Bletilla striata is a well-known traditional Chinese herb with varieties of functions. In China, the natural resources of Bletilla striata have been severely damaged because of the excessive exploitation and destruction of natural habitats. The aim of present study was to provide a reference for fully exploiting and utilizing the germplasm resources of Bletilla striata. The genetic diversity of 50 varieties of Bletilla striata from different area in China was analyzed by SCoT and IRAP molecular marker technique. A total of 209 bands were amplified by 20 groups of SCoT primers, of which 201 (96.17%) were polymorphic, and 47 polymorphic bands (94%) were observed in 50 bands amplified by 8 groups of IRAP primers. The 50 populations of Bletilla striata were divided into two major groups by SCoT and IRAP at the genetic similarity coefficient value of 0.60 and 0.68 individually. The partition of clusters in the unweighted pair group method with arithmetic mean dendrogram and principal coordinate analysis plot based on the SCoT and IRAP markers was similar. Results indicated the abundant genetic diversity of Bletilla striata among different areas. Our results will provide useful information for resource protection.

Ischemic acute kidney injury (AKI) is a prevalent disorder among hospitalized patients worldwide. Astragaloside IV (AS-IV) has been shown to protect against ischemic AKI. However, the specific effects and mechanisms of AS-IV on alleviating kidney ischemia-reperfusion (I/R) injury remain unclear. The objective of this research was to elucidate the regulatory targets and mechanisms through which AS-IV protects kidney I/R injury. A combination of network pharmacology, molecular docking, molecular dynamics (MD) simulation, pharmacodynamic study and Western blot were employed to explore the underlying mechanisms. Network pharmacology revealed that ferroptosis was a potential mechanism of AS-IV against kidney I/R injury. Molecular docking and MD simulations demonstrated strong binding affinity between the GPX4/SLC7A11 and AS-IV. The experimental verification demonstrated that AS-IV improved cell proliferation, decreased the level of ROS and Fe²⁺, and increased the expressions of GPX4 and SLC7A11 as same as Ferrostatin-1 in OGD/R-injured HUVECs. In conclusion, AS-IV had a significant inhibition on ferroptosis in kidney I/R injury, providing a new perspective for drug development on kidney I/R injury. Definitely, further exploration in vivo is necessary to fully understand whether AS-IV alleviates kidney I/R injury through inhibiting endothelial ferroptosis.

Introduction: Dysfunction of brain microvascular endothelial cells (BMECs) induced by oxidative stress represents a critical event in the pathogenesis of ischemia/reperfusion (I/R) injury. Although previous investigations have demonstrated the protective effects of Hydroxysafflor yellow A (HSYA) against I/R injury, the precise underlying mechanisms remain incompletely understood. Methods: An oxygen-glucose deprivation/reoxygenation (OGD/R) model was established in rat BMECs (rBMECs). We analyzed cell viability, proliferation, oxidative stress markers (SOD, JC-1), and pyroptosis-related protein expression (NLRP3, Caspase-1, GSDMD, IL-1β). The NLRP3 inhibitor MCC950 was utilized to elucidate HSYA’s regulatory role in OGD/R-induced pyroptosis. The network pharmacology approach was used to identify potential targets of HSYA against ischemia/reperfusion (I/R) injury. Molecular docking, molecular dynamics (MD) simulation, CETSA, DARTS assays, along with PARP-1 overexpression/inhibition experiments were performed to elucidate the underlying mechanism of HSYA in ameliorating I/R injury. Results: The results indicated that HSYA enhanced cell viability and proliferation of rBMECs exposed to OGD/R injury, accompanied by increased SOD activity and preserved MMP. HSYA suppressed the expression of pyroptosis-related proteins (NLRP3, Caspase-1, GSDMD, and IL-1β). The protective effects of HSYA were comparable to those observed with the NLRP3 inhibitor MCC950. Cotreatment afforded superior protection and more pronounced inhibition of NLRP3-mediated pyroptosis in OGD/R-induced rBMECs. Network pharmacology identified PARP-1 as a key target of HSYA against I/R injury. This interaction was validated through molecular docking and MD simulation, which revealed stable binding with high-affinity. Further experimental validation using CETSA and DARTS assays confirmed the direct binding of HSYA to PARP-1. Modulation of PARP-1 activity resulted in altered NLRP3 expression; Notably, both the PARP-1 inhibitor Olaparib and HSYA suppressed NLRP3, suggesting that the protective effects of HSYA may be attributed to direct targeting of PARP-1/NLRP3 pathway. Discussion: This study demonstrates that HSYA protects rBMECs against OGD/R injury by directly targeting PARP-1, thereby inhibiting the NLRP3-mediated pyroptosis pathway. These findings reveal a novel mechanism of HSYA in mitigating I/R injury and implicate PARP-1 as a promising therapeutic target. Nevertheless, several limitations should be considered. The precise molecular details between PARP-1 and the NLRP3 inflammasome pathway require further elucidation, and our findings remain to be validated using animal models of cerebral I/R injury.