Introduction

The microenvironment is the interstitial, tissue, organ, or humoral component of the cell, which is involved in constituting the environment of survival. The stability of microenvironment is an important condition for maintaining normal physiological activities. Abnormal changes of microenvironment components can cause pathological changes in cells and tissues.1 Under normal physiological function, the microenvironment of organ tissues affects the normal physiological activities of organ tissues. The thymus microenvironment regulates the phenotype and number of hematopoietic cells by regulating chemokines,2 bone marrow microenvironment has an important influence on the immune, hematopoietic and motor systems of the body.3 In addition, microenvironments exist in the lungs, pancreas, spleen and other organs, and play an important role in their physiology and pathology.4–6 In pathological conditions, the regulation of microenvironment also plays an important role in the occurrence and development of diseases, including tumor microenvironment, immune microenvironment, inflammatory microenvironment, and tissue damage microenvironment. In particular, the tumor microenvironment has become one of the targets of drug therapy in recent years.7,8 The regulation of microenvironment such as immunity, inflammation and tissue damage is also important for the occurrence and development of diseases, which directly or indirectly affects the disease process and the effect of drug therapy.

Medicinal plant is a plant used in medicine to prevent and treat diseases. The whole or part of Medicinal plant is used as Medicinal raw material, which is widely used in ethnomedicine. A lot of scientific research has been carried out on the monomer extracted from Medicinal plant. And widely used in clinical, such as paclitaxel, berberine and so on.9 In 1983, Pan BTd et al extracted exosomes for the first time from sheep reticulocytes in vitro.10 In 2009, Canal’s research group found that the intercellular fluid in the leaves of sunflower seedlings contained plant-derived exosome-like nanovesicles, it was first demonstrated that plants can secrete plant-derived exosome-like nanovesicles (PLDENs) into cells.11 Medicinal plant-derived exosome-like nanovesicles (MPLDENs) are extracellular vesicles secreted by cells of medicinal plants and are rich in proteins, nucleic acids, lipids and other bioactive substances,12 it has anti-inflammatory, anti-tumor, immunomodulatory and anti-oxidative stress effects.13,14 In the past 15 years, research on the application and mechanism of MPLDENs in the improvement and treatment of various diseases has become a research hotspot. From the different biological characteristics of MPLDENs, it is discussed that MPLDENs, as a new type of natural nanomaterial, have high biocompatibility and a series of functions such as drug delivery.15,16 In the past 15 years, research on the application and mechanism of MPLDENs in the improvement and treatment of various diseases has become a research hotspot. From the different biological characteristics of MPLDENs, it is discussed that MPLDENs, as a new type of natural nanomaterial, have high biocompatibility and a series of functions such as drug delivery.13,17 Compared with other extracts or monomers of medicinal plants, MPLDENs, as natural biological nanovesicles, have rich endogenous substances and active ingredients, and possess multiple therapeutic effects. Due to the biological characteristics of MPLDENs, MPLDENs have high lipid solubility. Therefore, MPLDENs significantly increase drug absorption and drug membrane permeability. Meanwhile, many related studies on MPLDENs have confirmed that MPLDENs are targeted to organs. The concentration of MPLDENs in the target organs is much higher than that in other organs, which is conducive to increasing the drug concentration in the target organs and can significantly enhance the therapeutic effect.

Due to the large number of research articles on MPLDENs, the extraction methods and administration methods of MPLDENs vary to some extent in different articles. Not only that, in the articles related to MPLDENs studying the same disease, the microenvironment and regulatory mechanisms involved in different articles also have certain differences. Therefore, this review comprehensively outlines the basic characteristics and extraction and identification methods of MPLDENs, summarizes the extraction methods of MPLDENs from different medicinal plant sources, and visually expounds the three main extraction methods. Meanwhile, the therapeutic effects of MPLDENs in diseases of various systems are classified. Meanwhile, the therapeutic effects of MPLDENs in diseases of various systems are classified, We mainly emphasize the impact of MPLDENs on cells, tissues and the microenvironment where diseases are located, and further summarize the specific mechanisms of MPLDENs regulation on the microenvironment. In addition, our goal is to explore how to apply the biological and therapeutic characteristics of MPLDENs and provide perspectives and ideas for further research on MPLDENs and the expansion of clinical application of MPLDENs.

Medicinal Plant-Derived Exosome-Like Nanovesicles Biological Characters and Characteristics of MPLDENs

MPLDENs are around 30–300 nm in size,18 The particle size and morphology of different MPLDENs have certain differences.19 The size of MPLDENs extracted from most of the Mosaic organs, such as tea leaves, honeysuckle flowers and aloe leaves, is small, with the particle size range below 150nm.20–22 The grain size of MPLDENs extracted from most of the root organs, such as polygala and ginseng, is larger, and the particle size ranges above 250nm.23–25 This is due to the subtle differences caused by the different compositions and structures of various organs in medicinal plants. The form of MPLDENs is small round or oval vesicles, and most MPLDENs have a typical saucer-like structure.26 The composition of MPLDENs mainly includes proteins, nucleic acids, lipids and other secondary metabolites, etc. Due to the differences in the growth environment of medicinal plants and autologous genes, the types and contents of proteins, nucleic acids and lipids in different MPLDENs are quite different, and the biological role played by MPLDENs is closely related to its composition. The researchers analyzed the proteins in MPLDENs and classified them into functional proteins, structural proteins and secreted proteins according to their functions.27,28 Functional proteins are involved in many biological processes such as cell expansion, protein folding, and heat stress.29,30 Wheat -derived PLDENs contain heat shock proteins and chaperone proteins that prevent heat stress in the rice during bad weather and help with protein folding.31 Nucleic acid is one of the main components of MPLDENs and plays an important role in the cell biology process. Due to the advantages of wide application and long-term effectiveness of miRNAs in treatment, miRNAs from MPLDENs have been widely extracted and identified and applied in the treatment and research of diseases.13 At the same time, MPLDENs can deliver siRNA by wrapping siRNA and thus play a role in disease treatment.32 Studies have shown that lipids, as the main components of MPLDENs, play a role in maintaining structural stability and participating in biological functions in vesicles. MPLDENs are phospholipid bilayer membrane structures, similar to the membrane structure of cell membrane, and contain a variety of lipid components, such as triglycerides, diacylglycerol, phosphatidylcholine and phosphatidylinositol.27,33,34 It has been reported that diacylglycerol and phosphatidic acid have a significant effect on membrane fusion and are involved in biological functions such as vesicle uptake, which is one of the advantages of exosome-like nanovesicles of medicinal plant origin into the human body35 (Figure 1).

Figure 1 Medicinal plant-derived exosome-like nanovesicles diagram.13,18,27,32–34

Methods for Extraction, Purification and Identification of MPLDENs

MPLDENs can be extracted and purified by a variety of methods, the main extraction and purification methods of MPLDENs include differential ultra-high speed centrifugation, sucrose gradient density centrifugation, immunoaffinity capture, ultrafiltration, size exclusion chromatography, PEG co-precipitation and so on,27 the extraction and purification methods of exosome-like nanovesicles from medicinal plants were summarized (Table 1). Among the MPLDENs extraction methods, differential ultra-high speed centrifugation and sucrose gradient density centrifugation are the most widely accepted.18 It is also the most used extraction and purification method. Three extraction methods steps (Figure 2). According to the biological properties and characteristics of MPLDENs, the morphology, particle size, protein concentration, nucleic acid content, protein type, nucleic acid type and lipid analysis of plant vesicles were evaluated.36–38 The identification methods of plant vesicles were summarized, and the advantages of different methods were compared (Table 2).39–46

Table 1 Extraction methods of MPLDENs from medicinal plants

Table 2 Comparison of plant vesicles identification methods

Figure 2 Methods for extraction and purification of MDPLNs (1) Method A: Differential ultra-high speed centrifugation; (2) Method B: Method A: Differential ultra-high speed centrifugation unite ultrafiltration; (3) Method C: Differential ultra-high speed centrifugation unite sucrose density gradient centrifugation method.

The Role and Mechanism of MPLDENs in Disease Treatment and Microenvironment Regulation

As a new type of therapeutic drug, MPLDENs enter the body through different modes of administration, so as treat the diseases occurring in each system in a targeted way, treating diseases by directly targeting disease tissues and cells and regulating the disease microenvironment. This section summarizes and discusses the therapeutic effect of MPLDENs on diseases of respiratory, digestive, motor, endocrine, circulatory, nervous and reproductive systems and skin diseases verified by in vivo and in vitro experiments and elucidates its mechanism of action on diseases and microenvironment regulation.

Respiratory System

MPLDENs are used in the treatment of respiratory diseases through different administration routes. The following diseases have been used in existing research Viral Pneumonia, Acute lung injury, Pulmonary fibrosis, Bronchial asthma (Figure 3 and Table 3).

Table 3 MPLDENs for the treatment of respiratory diseases

Figure 3 Mechanism of MPLDENs in the treatment of respiratory diseases.

Viral Pneumonia (VP)

VP is a kind of pulmonary inflammation caused by virus immersed in lung epithelial cells through respiratory tract. The main clinical manifestations are asthma, dry cough, fever and headache.84 Influenza A virus (IAV) and 2019 novel coronavirus (SARS-CoV-2) are the main types of viruses that cause VP in recent years.85,86

After the virus infects the body, it invades the lung and activates the immune system, recruits immune cells, transmits a large number of pro-inflammatory and anti-inflammatory cytokines, and shapes the pulmonary inflammatory microenvironment.87 In terms of improving the inflammatory microenvironment in the lungs, Houttuynia cordata-derived exosome-like nanovesicle miR168a-3p, miR168b-3p, novel_mir2 and miR8175 regulate the expression of PI3K-AKT pathway. novel_mir2 also regulates the expression of MAPK3.47 It can inhibit inflammation and improve the pulmonary inflammatory microenvironment. miR396a-5p from Ginger-derived exosome-like nanovesicles alleviates lung microenvironment inflammation by targeting the inhibition of NF-κb pathway mediated inflammation.68Taraxacum-derived exosome-like nanovesicles encapsulated PGY-sRNA-6 targeting the RELA gene in the NF-kB complex, reducing p65 protein expression, reducing oxidative stress, and inhibiting lung inflammation.47Pueraria lobata-derived exosome-like nanovesicles (PuLDENs) can regulate the balance of lung M1/M2 macrophages, increase the expression of M2 macrophages, improve the pulmonary inflammatory microenvironment, and reduce lung inflammation.48,56

In addition to improving the pulmonary inflammatory microenvironment, the proteins transcribed by the viral genome are closely related to the formation of the pulmonary microenvironment, and inhibiting the transcription of the viral genome is conducive to improving the pulmonary microenvironment.88 miRNAMIR2911 in Lonicera japonica Thunb-derived exosome-like nanovesicles targets multiple subtypes of IAV, including H1N1, H5N1, and H7N9. The expression of PB2 and NS1 proteins encoded by H1N1 was significantly reduced, thus significantly inhibiting the replication of H1N1 virus.82 In 2023, Zhu et al showed that miR858a and miR858b in Houttuynia cordata-derived exosome-like nanovesicles inhibited the expression of NP genes in IAV. miR166a-3p targets the inhibition of ORF1ab gene in SARS-CoV-2, reducing viral replication in vivo.47 At the same time, Ginger derived exosome-like miR396a-5p from nanovesicles inhibited the expression of viral S proteins and Nsp12 and Nsp13 proteins. The Cytopathic Effect (CPE) of SRAS-CoV-2 was inhibited in Vero E6 cells.68 This reduces the virus’s damage to cells and indirectly reduces the amount of virus in the body. In 2025,

The miRNAs in MPLDENs plays a major role in inhibiting virus replication, and miRNAs plays an important regulatory role in VP. The miRNAs produced in the body can inhibit virus infection symptoms and virus replication. In addition, viral infection can regulate the expression of miRNA in the body, regulate the pulmonary inflammatory microenvironment, and carry out immune escape through self-produced miRNA.89 Therefore, miRNAs in MPLDENs may not only inhibit viruses but also serve as endogenous miRNAs in the body to improve the pulmonary inflammatory microenvironment.

In conclusion, MPLDENs and their miRNAs can treat viral pneumonia by regulating the pulmonary inflammatory microenvironment and reducing the virus content in the body, preventing immune overactivation and increasing endogenous healing.90 Effective prevention of secondary pneumonia and acute respiratory distress syndrome.91,92 The two mechanisms are synergistic. Clinically, anti-inflammatory therapy and antiviral therapy are played by different drugs, and MPLDENs have a dual role, which can play a dual role and have a synergistic therapeutic effect. In addition to antiviral therapy, it can alleviate respiratory tract and fever symptoms such as asthma and dry cough in patients.

Acute Lung Injury (ALI)

ALI is the injury of alveolar epithelial cells and capillary endothelial cells caused by direct or indirect injury factors, which can lead to diffuse pulmonary interstitial and alveolar edema and acute hypoxic respiratory insufficiency, with high morbidity and high mortality.93 The damaged microenvironment formed by acute lung injury plays an important role in the development of ALI. By regulating the inflammatory microenvironment of ALI, M1 macrophages are reduced, M2 macrophages are increased, neutrophil infiltration is reduced, the secretion of proteins and inflammatory factors in lung tissue is inhibited, and the inflammatory microenvironment of ALI is improved, which can effectively alleviate ALI symptoms.94 In 2024, Ye et al found that Artemisia derived exosome-like nanovesicles (ALDENs) regulated the production of inflammatory factors and reduced inflammatory infiltration by inhibiting NF-κB and MAPK signaling pathways, and at the same time improved the mitochondrial function of macrophages by interacting with GABA receptors.55 Thus, it plays an anti-inflammatory role and prevents the progressive development of ALI. In 2023, Qiu et al showed that miR-7972 in fresh Rehmanniae Radix derived exosome-like nanovesicles extreme macrophages from M1 phenotype to M2 phenotype, reducing the production of inflammatory factors. miR-7972 inhibits Hedgehog pathway by targeting GPR161 protein and upstream and downstream GPR161 protein, thereby improving the inflammatory symptoms of ALI.60 By regulating the inflammatory microenvironment and immune microenvironment of ALI, MPLDENs and their miRNAs can alleviate the damage of alveolar epithelial cells and capillaries, control diffuse inflammation, and reduce pulmonary interstitial and alveolar edema. Fu et al showed that Platycodon grandiflorum-derived exosome-like nanovesicles can alleviate acute lung injury caused by IAV infection by regulating the mechanisms of immune and metabolic anti-inflammatory damage.81

Pulmonary Fibrosis (PF)

PF is a chronic interstitial lung disease characterized by abnormal activation of fibroblasts, excessive accumulation of extracellular matrix, inflammatory damage and abnormal lung remodeling.95 Hong Jing Tian-derived exosome-like nanovesicles and Taraxacum-derived exosome-like HJT-sRNA-m7 in nanovesicles (taraxion) reduce collagen and fibrin production by fibroblasts by inhibiting gene expression of α-SMA, Fibronectin (FN), and collagen type I α1 (COL1A1)48,96 and control the progressive development of pulmonary fibrosis. HJT-sRNA-m7 maintains extracellular matrix microenvironmental homeostasis and reduces epithelial mesenchymal transformation by reducing fibrin accumulation, thereby inhibiting myofibrocyte formation in lung mesenchyma,94,95 preventing fibroblasts from being over-activated and delaying pulmonary fibrosis.

Bronchial Asthma (BA)

BA is a chronic inflammatory disease of the airway, which is accompanied by damage and detachment of the airway epithelia, airway hyperresponsiveness and increased mucosal secretion, leading to symptoms such as cough, wheezing, shortness of breath, and eventually airway remodeling.96Aster yomena Callus-derived exosome-like nanovesicles (ACLDENs) have a protective effect on allergic airway diseases by inhibiting the T cell response associated with asthma and increasing the level of regulatory T cells (Foxp3+CD25+CD4+). Inhibition of airway overimmune response, thereby relieving airway smooth muscle tension and spasm, reduce airway hyperreactivity. Aster yomena callus-derived exosome-like nanovesicles inhibit Th2 and mixed Th2/Th17 responses associated with asthma, reduce helper T cell infiltration, regulate the immune microenvironment of asthmatic97 and reduce mucosal secretion. Improve symptoms such as cough, wheezing, and shortness of breath.61

Digestive System

MPLDENs are used in the treatment of digestive system through different administration routes. The following diseases have been used in existing research: ulcerative colitis, digestive system tumors, acute liver failure (Figure 4 and Table 4).

Table 4 MPLDENs for the treatment of digestive system

Figure 4 Mechanism of MPLDENs in digestive system.

Ulcerative Colitis (UC)

UC is a chronic inflammatory disease that affects the function of the colon, with typical symptoms including diarrhea, rectal bleeding, and abdominal pain. MPLDENs treat ulcerative colitis by improving intestinal inflammation with immune microenvironment and regulating intestinal microenvironment.

In terms of MPLDENs improving intestinal inflammation and immune microenvironment, in 2019, Zhu et al‘s research shows that Portulaca oleracea-derived exosome-like nanovesicles (PoLDENs) down-regulated the expression of Zbtb7b in conventional CD4+ T cells through the Lactobacillus-Enterobacteriaceae-indole derivative axis.97 Zbtb7b plays a role in peripheral CD4+ T cells, inhibiting the expression of CD8 lineage genes, including the expression of CD8 and cytotoxic effector genes perforin and granzyme B, and inhibiting the expression of IFN-γ during effector differentiation.97 The conventional CD4+ T cells are reprogrammed into DP CD4+CD8+ T cells (DP cells), thereby regulating the immune microenvironment of ulcerative colitis, inhibiting the production of TNF-α, IL-1β, IL-6, IL-12 and other inflammatory factors, increasing the production of anti-inflammatory factors, and significantly alleviating the symptoms of colitis and has specific targeted therapeutic action.97 Edible pueraria lobata-derived exosome-like nanovesicles (PuLDENs) reduce the formation of M1-type macrophages and regulate the immune microenvironment by polarizing macrophages into M2 macrophages. Reduce the production of TNF-α, IL-1β, IL-6 and other inflammatory factors, reduce intestinal epithelial damage, promote the proliferation of intestinal epithelial cells and restore barrier function,56 thereby reducing the inflammatory response of UC and repairing tissue damage caused by UC. In 2022, Liu et al showed that Turmeric-derived exosome-like nanovesicles (TLDENs) improved mouse colitis and accelerated the resolution of colitis by regulating inflammatory factors and antioxidant gene HO-1, and the potential mechanism of its treatment may be related to the NF-κB pathway.70

TLDENs can not only alleviate the symptoms of ulcerative colitis by improving the intestinal inflammatory microenvironment, but Gao et al found that TLDENs can also promote the transformation of M1 macrophages into M2 macrophages, up-regulate the expression of tight junction protein and regulate the intestinal microbiota to restore the damaged intestinal epithelial barrier and protect the intestinal barrier function.27 The effects of TLDENs inhibited intestinal inflammation and reduced intestinal damage. Sophora japonica exosome-like nanovesicles reduce iron death in intestinal epithelial cells and regulate metabolic disorders by regulating HIF pathway, thereby repairing intestinal mucosa, and have potential therapeutic effects on ulcerative colitis.75 Ginseng-derived exosome-like nanovesicles (GinLDENs) improve the immune microenvironment of ulcerative colitis by regulating the polarization of macrophages, inhibiting the anti-inflammatory effect of NF-κB pathway.74 Zhao et al found that Garlic derived exosome-like nanovesicles (GaLDENs) treated ulcerative colitis by inhibiting the TLR4/MyD88/NF-κB signaling pathway, thereby inhibiting downstream cascade reaction and the release of inflammatory factors. Inhibition of TLR4 expression may act through Han-miR3630-5p in GaLDENs.50 In 2022, Sriwastva et al found that Mulberry bark-derived exosome-like nanovesicles (MBLDENs) activate the anti-inflammatory pathway mediated by AhR-COPS8, AhR signal transduction through the combination of MBLDENs derived heat shock protein HSPA8 (MBELN-HSPA8) and AhR to activate and improve the inflammatory microenvironment of colitis and alleviate colitis symptoms.73

In the regulation of intestinal microenvironment by MPLDENs, garlic-derived exosome-like nanovesicles (GaLDENs) can restore the relative abundance of Lachnospiraceae beneficial to intestinal microenvironment. Decreased the abundance of Helicobacter, which leads to chronic intestinal inflammation97 peu MIR2916-p3 in GaLDENs specifically promoted the growth of Bacteroides thetaiotaomicron, which is beneficial to the intestinal microenvironment.71 To change the intestinal microbiota spectrum of colitis mice, which has a preventive effect on the occurrence of ulcerative colitis. Ginseng-derived exosome-like nanovesicles (GinLDENs) reduce the ratio of Firmicutes/Bacteroides, regulate intestinal flora, and slow the progression of ulcerative colitis.74 Fresh Rehmannia exosome-like nanovesicles can reduce the abundance of Escherichia coli and regulate the microenvironment of intestinal flora by inhibiting the formation of Escherichia coli biofilms.60

In conclusion, MPLDENs can regulate the balance of M1 and M2 macrophages and CD4+ T cell genes to regulate the immune microenvironment, reduce the production of inflammatory factors and increase the expression of pro-inflammatory factors, thus inhibiting inflammatory response, improving the inflammatory microenvironment, regulating the type and abundance of intestinal flora, and optimizing the intestinal microenvironment for the treatment of ulcerative colitis. Relieve symptoms such as diarrhea, abdominal pain and blood in the stool, and repair the intestinal barrier effect, reducing the damage of ulcerative colitis to the colon. Intestinal microflora microenvironment, immune microenvironment and inflammatory microenvironment are closely related to the progression and treatment of ulcerative colitis. The immune microenvironment improves the intestinal microflora microenvironment and inflammatory microenvironment, and the intestinal microflora and inflammatory microenvironment affect the expression and function of macrophages and dendritic cells in the immune microenvironment.99

Digestive System Tumors

Colorectal cancer is a malignant tumor occurring in the colon or rectum. Early clinical manifestations are usually hematochezia, diarrhea and abdominal pain, and late symptoms are anemia and cachexia with weight loss.100,101 Ginseng-derived exosome-like nanovesicles (GinLDENs) enhance the efficacy of immune checkpoint antibodies by reprogramming the cold tumor microenvironment, thereby promoting T lymphocyte infiltration and anti-tumor response. Improved the efficacy of immune checkpoint inhibitor (ICI) related combination in the treatment of colon cancer.75 Ginger-derived exosomes like nanovesicles (GLDENs), after being taken up by colorectal cancer cells, mediated the metabolic transformation of cancer cells in the tumor microenvironment by activating the PI3K-AKT signaling pathway, and PI3K/AKT signaling was associated with apoptosis of cancer cells.102 Promote the apoptosis of colorectal cancer cells and inhibit the cell viability of colorectal cancer cells and have anti-colorectal cancer effect.98

Hepatocellular carcinoma is a malignant tumor occurring in the liver, which often occurs in the terminal stage of chronic liver disease and decompensated stage of cirrhosis.103 Oral administration of Morus nigra L. leaves-derived exosome-like nanovesicles (MLLDENs) significantly inhibited tumor growth in mouse hepatoma models. After MLDENs intervention with Hepa1-6 cells, the cell cycle arrest in G0/G1 phase of Hepa1-6 cells and apoptosis of Hepa1-6 cells were induced.97Asparagus cochinchinensis-derived exosome-like nanovesicles (ALDENs) can induce apoptosis of Hep G2 cells by up-regulating protein levels of AIF, Bax and Bak and activating caspase-9, inhibiting the occurrence, development and metastasis of liver cancer.97 However, apoptosis of liver cancer cells affects the inflammatory microenvironment of liver cancer, thus accelerating the progression of tumor.104 After the apoptosis of liver cancer cells is induced by MLDENs and ALDENs, the inflammatory response of the liver is significantly reduced, indicating that in addition to promoting the apoptosis of tumor cells, the tumor inflammatory microenvironment can be improved and inflammatory infiltration can be reduced. MLDENs and ALDENs both target the liver and have good biosafety, and the inflammation and liver function indexes are close to normal. In the absence of damage to normal liver cells, better targeted treatment of liver cancer.72,79

In conclusion, MPLDENs can induce apoptosis of cancer cells by increasing PI3K/AKT signal and activating caspase-9 to slow down tumor growth. The cold tumor microenvironment can be changed into a hot tumor microenvironment, which can increase the efficacy of immunotherapy and improve the tumor inflammatory microenvironment to alleviate some series of symptoms such as fever and pain caused by tumors. Digestive system tumors, especially liver cancer, have a high degree of malignancy, easy to metastasize, and the symptoms of fever and pain caused by tumors are more serious. MPLDENs treat both the primary tumor and the adverse symptoms of tumor-induced inflammation.

Acute Liver Failure

Acute liver failure, also known as fulminant liver failure, is a rare life-threatening disease with major complications including type A hepatic encephalopathy and multiple organ failure, and a high mortality rate.105,106 Garlic-derived exosome-like nanovesicles (GaLDENs) inhibit the production of inflammatory factors, inhibit CCR2/CCR5 signaling, and reduce the aggregation of macrophages, monocytes and other immune cells into the liver. Reducing the intense immune response and inflammatory response caused by acute liver failure,66 regulating the inflammatory microenvironment, GaLDENs as a natural plant extract has high biosafety, low liver toxicity, alleviating heavy liver metabolic burden, and controlling the process of acute liver failure.50

Motor System

MPLDENs is used in the treatment of digestive system through different administration routes. The following diseases have been used in existing research osteoporosis, osteoarthritis, osteosarcoma, muscle atrophy (Figure 5 and Table 5).

Table 5 MPLDENs for the treatment of motor system diseases

Figure 5 Mechanism of MPLDENs in muscle atrophy.

Osteoporosis

Osteoporosis is a systemic bone disease caused by a variety of reasons, the main characteristics are the decrease of bone density and bone quality, the damage of bone microstructure, the increase of bone fragility, the high probability of fracture, and the reduction of bone trabeculae.108,109 MPLDENs treatment of osteoporosis works through its effects on osteoblasts and osteoclasts, thereby regulating dysfunction of bone cell activity and abnormal cytokine secretion in the osteoporosis microenvironment, reducing bone mass loss and bone imbalance.110Pueraria lobata-derived exosome-like nanovesicles (PuLDENs) can improve the differentiation and calcification of hBMSCs cells, regulate the imbalance of intestinal flora through TMAO, and increase the autophagy of BMSC cells to promote bone formation and increase the number of osteoblasts. Improve and treat osteoporosis.57Rhizoma Drynariae-derived exosome-like nanovesicles (RLDENs) induce osteoblast differentiation by activating ERα signaling pathway, up-regulating Runx2 and BMP2. RLDENs can effectively relieve bone loss in vivo, maintain normal bone density and bone quality, and prevent and treat osteoporosis.62Morinda Officinalis-derived exosomes like nanovesicles (MoLDENs) increased the expression of CREB and RSK1 proteins and promoted the proliferation of MC3T3-E1 cells by activating MAPK signaling pathway. MC3T3-E1 cells were induced to differentiate into osteoblasts to reduce bone trabecular destruction, increase bone density and alleviate osteoporosis.67 Yam-derived exosome-like nanovesicles (YLDENs) also promoted the proliferation and differentiation of MC3T3-E1 cells by activating MAPK signaling pathway and up-regulating p38.80 Ginseng-derived exosome-like nanovesicles (GinLDENs) inhibited osteoclast differentiation and reduced bone microstructure damage by inhibiting IκBα, JNK and ERK signaling pathways and down-regulating c-Fos, c-Jun and NFATc1 genes. Delay the progression of osteoporosis.76 By promoting the proliferation and differentiation of osteoblasts and inhibiting the differentiation of osteoclasts, MPLDENs can alleviate and treat osteoporosis in various aspects, maintain bone density, bone quality and the number of bone trabeculae, reduce the damage of bone microstructure, and maintain a bone microenvironment with a balance between osteoclasts and osteogenesis.

Osteoarthritis

Osteoarthritis is a non-inflammatory degenerative joint disease characterized by pain and stiffness in the joints.97 Garlic derived exosome-like nanovesicles (GaLDENs) regulate inflammatory signaling pathways, improve inflammatory microenvironment, and relieve pain symptoms by inhibiting MAPK signaling pathway and phosphorylation of ERK, JNK, and P38. GaLDENs promote the expression of collagen and agglutinoglycan, play a role in cartilage repair, improve the symptoms of bone and joint stiffness, and slow down the progression of osteoarthritis.97

Osteosarcoma

OS is the most common primary bone malignancy with a high tendency of local invasion and metastasis.97 The main symptoms are pain, mass, lameness, and related systemic symptoms.97 The tumor immune microenvironment has been shown to promote cancer cell proliferation, migration, apoptosis, and drug resistance, as well as immune escape.97Dipsacus Asperoides-derived exosome-like nanovesicles (DLDENs) can significantly inhibit the invasion and migration of osteosarcoma cells by activating the low-toxicity and high-affinity apoptosis pathway induced by P38/JNK signal. Inhibit the proliferation of osteosarcoma, so as to achieve the role of treatment of osteosarcoma.97

Muscle Atrophy

Muscular dystrophy refers to the reduction of muscle volume caused by rhabdomyodystrophy, the thinning or disappearance of muscle fibers, and the main symptoms are loss of normal muscle strength, fatigue and reduced exercise capacity, and reduced quality of life.97 Gouqi-derived exosome-like nanovesicles (GouLDENs) activation of AMPK/SIRT1/PGC1α signaling pathway can improve quadriceps grip strength and increase the muscular area, change the metabolites of quadriceps muscle, and regulate the internal microenvironment of C2C12 cells. Promotes muscle regeneration in mice with muscle atrophy.97

Endocrine System

MPLDENs are used in the treatment of endocrine system through different administration routes. The following diseases have been used in existing research type 2 diabetes, diabetic ulcer (Figure 6 and Table 6).

Table 6 MPLDENs for the treatment of circulatory, endocrine, nervous and reproductive systems

Figure 6 Mechanism of MPLDENs in circulatory, endocrine, nervous and reproductive systems.

Type 2 Diabetes

Type 2 diabetes is a chronic disease caused by insufficient or inefficient use of insulin, also known as insulin-dependent diabetes, the main symptoms of polydipsia, polyphagia and polyuria.118 Mung Bean Sprouts-derived exosomes -like nanovesicles (MLDENs) increased the expression of P-GSK-3β and GS by regulating the PI3K/Akt/GLUT4/GSK-3β pathway, which promoted glycogen synthesis and glycogen synthesis and transformation. Converting glucose into glycogen storage, GSK-3β is a protein that regulates Nrf2 oxidative stress level,119 promotes the expression of Nrf2 factor, reduces the oxidative stress response caused by insulin resistance in hepatocytes, restores the normal physiological function of hepatocytes, and increases the expression of GLUT4 protein in insulin-resistant hepatocytes. Promote glucose absorption and glycogen synthesis, reduce blood sugar, increase glucose tolerance.97 Insulin resistance is closely related to the pathogenesis of type 2 diabetes.97 Insulin resistance causes compensatory production of insulin by functional islet beta cells, disrupts the microenvironment that maintains the islets, and even leads to islet failure.121 Garlic-derived exosome-like nanovesicles (GaLDENs) can reverse insulin resistance97 by increasing Acinetobacter mucophilus and stimulating GLP-1 signaling pathway to reverse insulin resistance51 and protect existing islet beta cells. Maintenance of islet microenvironment balance.123Astragalus-derived exosome-like nanovesicles (ALDENs) can improve glucose metabolism and reduce blood sugar by regulating the ratio of Firmicutes and Bacteroidetes and their metabolites.64 Type 2 diabetes is associated with complications as the disease progresses.124 Citrus reticulatae Pericarpium-derived exosome-like nanovesicles can enhance the expression of FAO-related genes and inhibit the expression of genes associated with liver lipogenesis and glycolysis by regulating intestinal flora. Modulating liver lipid metabolism CiLDENs activate FXR-SHP pathway and promote liver bile acid effection, inhibit cholesterol accumulation and alleviate T2DM induced hepatic steatosis.54

Diabetic Ulcer

Diabetic ulcer is a complication of diabetic peripheral neuropathy, somatic nerve and sympathetic autonomic neuropathy. Diabetic ulcers have extremely complex microenvironments, such as hyperglycemia, ischemia, hypoxia, excessive inflammation, persistent infection, high protease activity, and insufficient energy supply.97 Non-healing of diabetic chronic ulcers is usually due to impaired angiogenesis.97 Ginseng-derived exosome-like nanovesicles (GinLDENs), through accelerating the biological process of glycolysis, provide substances and energy for the repair and formation of blood vessels, inhibit oxidative phosphorylation, reduce oxidative stress, and restore angiogenesis in diabetic ulcers. Improving the microenvironment of diabetic ulcer in many ways and controlling the progress of diabetic ulcer.77

Circulation System

Myocardial injury is a serious pathological consequence of circulatory system.127 Drug-induced myocardial injury is a common problem in clinical practice.97 Mitochondrial dysfunction and mitochondrial injury of cardiomyocytes are the main causes of death of cardiomyocytes and endothelial cells in myocardial injury.129,130Momordica charantia L.-derived exosomes like nanovesicles (McLDENs) regulate the p62/Keap1/Nrf2 pathway to reduce the increased cardiotoxicity and apoptosis of cardiomyocytes induced by doxorubicin (DOX) during its antitumor effects.131 Panax ginseng-derived exosome-like nanovesicles (PGinLDENs) improved abnormal apoptosis of cardiomyocytes by regulating apoptosis factors, increasing the expression of anti-apoptotic factor Bcl-2, inhibiting the expression of pro-apoptotic factors Bax and Cyt C. ameliorating myocardial damage caused by DOX.116 Oxidative stress is an important pathological mechanism of myocardial injury.132 McLDENs can alleviate oxidative stress and promote the proliferation of DOX-treated cardiomyocytes by increasing the expression of Nrf2 factor and HO-1 gene.97 By regulating apoptosis, energy metabolism and oxidative stress of cardiomyocytes, MPLDENs protects cardiomyocytes, maintains normal physiological functions of cardiomyocytes, and improves pathological manifestations of myocardial injury. At the same time, McLDENs and PLDENs can both reduce mitochondrial damage and maintain normal physiological activities of mitochondria and reduce myocardial damage caused by DOX (Figure 6 and Table 6).

Nervous System

In neurological diseases, MPLDENs can penetrate the blood–brain barrier and delay the progressive exacerbation of neurological diseases by improving the energy metabolism of nerve cells, regulating the immune and inflammatory microenvironment. Pueraria lobata-derived exosome-like nanovesicles (PuLDENs) activate the Pink1-Parkin pathway, and PINK1 and Parkin control the specific elimination of dysfunctional or redundant mitochondria. Thus regulating the mitochondrial network and preserving energy metabolism,133 mediating mitochondrial autophagy, reducing dysfunctional mitochondria, restoring ATP supplementation, maintaining mitochondrial homeostasis thereby improving the cellular microenvironment, improving mitochondrial dysfunction in dopamine neurons,58 improving tremor and retardation caused by Parkinson’s disease in progressive neurodegenerative diseases.134 Polygalae Radix-derived exosome-like nanovesicles can cross the blood–brain barrier 25 and improve the manifestation of dementia caused by Alzheimer’s disease.135 Ginseng-derived exosome-like nanovesicles (GinLDENs) can directly act on glioma through the blood–brain barrier, promote apoptosis of glioma cells, inhibit cell growth of glioma cells, fibroblasts and endothelial cells, and act on glioma.136 Tumor cells actively reshape their microenvironment and create crosstalk between components of the microenvironment to promote disease development.137 By regulating tumor inflammation and immune microenvironment, GinLDENs can inhibit the inflammation of tumor tissue, regulate macrophages, inhibit M2 macrophages, reduce the expression of growth factors, angiogenic factors and chemokines, and slow down the occurrence and development of glioma78 (Figure 6 and Table 6).

Reproductive System Cervical Cancer

Cervical cancer is a malignant tumor occurring in the female reproductive tract, which occurs in the cervix. Human tumor virus (HPV) infection is the main cause of cervical cancer.138,139 HPV-infected cells actively reshape the local environment by conspiring with cells and inducing a supportive and immunosuppressive post-infection microenvironment that supports virus persistence, transmission, and promotes cervical cancer97 MiR2911 from Lonicera japonica Thunb-derived exosome-like nanovesicles binds to the E6 and E7 oncogenes of HPV16/18 to inhibit the proliferation of cervical cancer cells. The apoptosis of cervical cancer cells was induced by regulating Caspase3 to achieve the purpose of treating cervical cancer.20

Breast Cancer

Breast cancer is a disease in which abnormal breast cells grow out of control and form tumors.97Brucea javanica-derived exosome-like nanovesicles (BLDENs) inhibit the PI3K/Akt/mTOR pathway, and the PI3K/Akt/mTOR pathway, PI3K/AKT signal mediates the metabolic transformation of cancer cells in the tumor microenvironment97 resulting in the apoptosis of 41T tumor cells. At the same time, by inhibiting the synthesis of VEGF, inhibiting angiogenesis, improving the tumor microenvironment, reducing the blood flow of tumor tissue, and slowing down the development of breast cancer97 (Figure 6 and Table 6).

Skin Diseases

MPLDENs are used in the treatment of Skin Diseases through different administration routes. The following diseases have been used in existing research skin melanoma, skin trauma, skin aging, Alopecia (Figure 7 and Table 7).

Table 7 MPLDENs for the treatment of Skin disease

Figure 7 Mechanism of MPLDENs in skin disease.

Skin Melanoma

Cutaneous melanoma is a malignant tumor caused by melanocytes of the skin,143 Ginseng-derived exosome-like nanovesicles (GinLDENs) induced macrophage polarization by mediating the activation of TLR4-MyD88 pathway, accelerated the apoptosis of melanoma cells and the expression of apoptosis gene caspase 3/7, and inhibited the growth of melanoma.28

Skin Trauma

Skin trauma forms a damaging microenvironment that triggers inflammatory and immune responses.97 MPLDENs can repair wounds and improve local inflammatory symptoms. Wheat-derived exosome-like nanovesicles (WLDENs) accelerate wound repair by increasing type I collagen, promote the proliferation and migration of endothelial cells, epithelial cells and dermal fibrocytes, promote the repair of skin wound tissue and the formation of blood vessels, and improve the skin injury microenvironment.53Dendrobium-derived exosome-like nanovesicles (DLDENs) improve the inflammatory microenvironment and promote wound healing by inhibiting overexpression of immune cells and genes.97

Skin Aging

Skin aging is more pronounced with age because senescent cells accumulate with age, and during skin aging, the extracellular matrix microenvironment undergoes tremendous structural changes and degradation, resulting in thinning of the dermis and loss of elasticity in the aging phenotype.134,145 As a sign of aging, cell senescence is a basically irreversible cell cycle arrest with the characteristics of apoptosis.146Bletilla-derived exosome-like nanovesicles (BLDENs) inhibited the apoptosis of human immortalized epidermal cells by regulating caspase-9 and Caspase-E3 in the Caspase-dependent apoptotic pathway.160 BLDENs avoid cell cycle arrest and optimize extracellular matrix microenvironment, slow down skin aging.135,146

Alopecia

Alopecia refers to the phenomenon of hair loss. There are many causes of hair loss, such as androgenic alopecia, endocrine alopecia and physical alopecia.97 Studies have shown that the epidermis and perihair follicle axons within the intact hairy skin of mice have significant dynamic plasticity related to their microenvironment.136 By applying Garlic derived exosome-like nanovesicles (GaLDENs), the Wnt/β-catenin signaling pathway can be regulated, the hair follicle microenvironment can be improved, the release of growth factors can be increased, the hair follicle development can be stimulated, and hair loss can be prevented or treated.97

MPLDENs as Advantageous Drugs for Microenvironment Regulation

In vivo imaging experiments have shown that MPLDENs can be used to target therapeutic organs. For example, Morinda and rhizome can target bone tissue, and puerariae can target cerebral cortex through the blood–brain barrier.57,62,67 Due to the many side effects of drugs in the treatment of some diseases and the serious damage to normal tissues, the research and development of targeted therapies and targeted drugs are now widely carried out in clinical and scientific research.147,148 The natural targeting of MPLDENs can bring non-negligible advantages for targeted treatment of diseases. It can be enriched in the disease site and play a therapeutic role. MPLDENs play a strong role in regulating the microenvironment where the disease is located. For the treatment of the disease microenvironment, MPLDENs can cooperate with symptomatic treatment and etiological treatment, consolidate the efficacy of other drugs for the disease, delay the development of the disease, and achieve better therapeutic effects.

MPLDENs have different modes of administration in different therapeutic systems, the main purpose of which is to make MPLDENs more effective. In the respiratory system, MPLDENs can be treated by inhalation, in the digestive system, mostly by oral administration, and in the circulatory system, it can be treated by intravenous injection. Skin diseases such as skin wounds are treated by external application. The microenvironment of a disease is composed of various factors such as the disease and the organism. When MPLDENs are used as the regulatory medium of the microenvironment, the mode of administration of MPLDENs determines the regulatory ability and therapeutic effect of MPLDENs to a certain extent, as well as affects the medical compliance of patients.147,149 As lipids are the main components of MPLDENs, the membrane structure of MPLDENs is phospholipid bilayer membrane structure, which is similar to that of cell membrane, and it is easy to fuse with the cell membrane to release the functional substances in MPLDENs. MPLDENs can penetrate into the tissue microenvironment, immune microenvironment, inflammatory microenvironment, extracellular matrix, etc., and has a high degree of biocompatibility, which not only increases the regulation of MPLDENs on the microenvironment, but also reduces the adverse reactions and toxicity caused by drugs in vivo.97

MPLDENs can be used as drug carriers to load drugs, MPLDENs, as non-cellular nanoscale structures, can deliver drugs for shuttle communication between cells, and MPLDENs can enter cells through the close connection between cells. The membrane lipid composition and surface proteins of MPLDENs can be further regulated to enhance their affinity and selectivity and improve their targeting properties. MPLDENs are able to protect loaded drugs from early degradation or clearance until they reach the target cell. Inside the cell, due to changes in the intracellular environment, drugs can be efficiently released from MPLDENs, producing efficacy. After MPLDENs are modified, MPLDENs can achieve better delivery results. In the engineering process, the way of drug administration and the location of drug delivery are important factors to consider. When drugs are orally absorbed through the digestive tract, the acid resistance of MPLDENs membrane surface protein needs to be taken into account. Sesame leaf-derived exosome-like nanovesicles as a delivery vector for luteolin maintain structural integrity and improve bioaccessibility during simulated digestion.151 When drugs are absorbed percutaneous by external application, the affinity between the lipids on the surface of MPLDENs membrane and the skin needs to be considered. Due to metabolic pathways such as first-pass effect, the original drug content to reach the diseased tissue is low. However, after the drug is loaded in MPLDENs engineered, the drug uses the engineered MPLDENs as the carrier, and the dose of the drug is smaller than that of the original drug, but the effect is the same as that of the original drug dose, so as to reduce the adverse reactions and metabolic burden caused by high-dose drugs.

All in all, due to the good organ-targeting, diverse drug delivery modes and high biocompatibility of MPLDENs, the regulation of MPLDENs on the disease microenvironment has been enhanced, which can alleviate disease symptoms and delay disease progression. After MPLDENs are engineered, MPLDENs can cooperate with other drugs to carry out disease treatment, achieve accurate and targeted localization of lesion sites, improve drug efficacy and reduce drug metabolic burden.

Future Prospect and Ch