Telomerase reverse transcriptase (TERT) is an essential component of telomerase that plays a significant role in regulating telomerase activity and maintaining chromosomal telomere length (Forino et al., 2025; Wyatt et al., 2010). In addition to its canonical role in telomere maintenance, the non-canonical functions of TERT have gained increasing recognition in recent research (Ale-Agha et al., 2014, Cuevas et al., 2025). Our team has extensively summarized and analyzed the critical role of TERT in neurological diseases (Li et al., 2010) and has continuously explored its potential role in hypoxic-ischemic brain damage (HIBD) in neonatal animals (Li et al., 2011). Our previous studies found that TERT reduces neuronal apoptosis, promotes neurogenesis and angiogenesis, and improves neurological function after HIBD (Li et al., 2013, 2022a, 2022b; Zhao et al., 2012), thereby exerting neuroprotective effects. However, in our previous studies (Li et al., 2022a, 2022b), TERT expression in neonatal animals was successfully enhanced through lentiviral or adenoviral infection approaches. While these studies established the therapeutic potential of TERT upregulation, the clinical translation of viral-based gene delivery faces substantial challenges, including immunogenicity risks in human neonates. Therefore, the current study investigates a clinically feasible pharmacological approach to enhance TERT expression following neonatal HIBD, which may offer a safer and more practical therapeutic alternatives for treating hypoxic-ischemic encephalopathy (HIE).

Astragalus membranaceus [The plant name Astragalus membranaceus (Fisch.) Bunge has been verified with The Plant List (www.theplantlist.org), accessed on March 28, 2025] is a widely used traditional Chinese medicine. Since the Qing Dynasty (the 17th century), large doses of Astragalus membranaceus have been used to treat stroke (Lo et al., 2021). Numerous studies have reported the neuroprotective effects of Astragalus membranaceus in ischemia-reperfusion brain injury (Liu et al., 2013; Lo et al., 2021), including reducing cell apoptosis and cerebral infarction. The protective effects of Astragalus membranaceus in neonatal HIBD have also been reported (Jia et al., 2003, 2005), but the specific mechanisms remain unclear. TA-65 is a commercially available small-molecule telomerase activator (Bawamia et al., 2023), composed of a >95 % pure single chemical entity isolated from a proprietary extract of the dried root of Astragalus membranaceus, along with inert excipients (Harley et al., 2011). TA-65 has been marketed for its potential health benefits (Liu et al., 2017; Salvador et al., 2016). The protective effects of TA-65 have been observed in both basic and clinical research on cardiovascular diseases (Ale-Agha et al., 2021; Maier et al., 2020) and are achieved by activating TERT expression (Ale-Agha et al., 2021; Bernardes de Jesus et al., 2011). In neurology, animal experiments have shown that TA-65 increases TERT expression in brain tissue following traumatic brain injury and promotes neurological function recovery after brain injury (Eyolfson et al., 2020). Research on mouse models of Parkinson's disease has shown that TA-65 upregulates TERT expression and activates autophagy, thus delaying disease progression (Wan et al., 2021). Therefore, TA-65 presents a promising application for TERT-based therapies in treating HIBD.

The primary objective of this study was to evaluate the therapeutic potential and safety profile of TA-65 for neonatal HIBD. To achieve this, we established both in vivo and in vitro models of HIBD in neonatal mice and administered TA-65 to: (1) systematically characterize its effects on TERT expression in brain tissue and neuronal cells; (2) quantify its neuroprotective efficacy through comprehensive assessment of brain injury and neurological function; and (3) conduct crucial safety evaluations to support future clinical translation for treating hypoxic-ischemic encephalopathy (HIE). This investigation provides the first direct evidence supporting TA-65 as a promising pharmacological strategy for HIE.