壮腰通络方调节SIRT1/HIF-1α信号通路抑制大鼠髓核细胞衰老延缓椎间盘退变的实验研究OA

Objective:Based on the theory of"invigorating the kidney and activating blood circulation",this study aims to investigate the mechanism by which Zhuangyao Tongluo for-mula(ZYTLF)delays intervertebral disc degeneration(IVDD)by inhibiting nucleus pulposus cell(NPC)senescence through regulation of the SIRT1/HIF-1α signaling pathway.Methods:Seventy 8-week-old SPF male rats were randomly divided into six groups:blank group,model group,positive drug control group(Celecoxib),and Zhuangyao Tongluo for-mula low-dose,medium-dose,and high-dose groups.Except for the blank group,a lumbar instability-induced IVDD model was established in all other groups by resection of the spinous processes,articular processes,and supraspinous ligaments.Intragastric administration was initiated on the third day after modeling and continued for 8 weeks.Behavioral perform-ance was observed,and intervertebral disc and nucleus pulposus tissues were collected.Hematoxylin-eosin(HE)staining and Safranin O-Fast Green(SO/FG)staining were used to observe the morphology of the intervertebral disc tissues.Western Blot and real-time quantitative PCR(RT-qPCR)were employed to detect the protein and mRNA expression lev-els of SIRT1,HIF-1α,p16,and Ki-67 in the nucleus pulposus tissues.Results:Compared with the blank group,the nucleus pulposus tissues in the model group exhibited shrinkage and aggregation,disordered arrangement of the annulus fibrosus,and severe loss of matrix proteoglycans.The protein and mRNA expression levels of HIF-1α and p16 were significantly in-creased(P＜0.05 or P＜0.01),while the protein and mRNA expression levels of SIRT1 and Ki-67 were decreased;how-ever,the difference in Ki-67 mRNA expression was not statistically significant(P＞0.05).Compared with the model group,the pathological damage in all ZYTLF intervention groups was alleviated to varying degrees,with the medium-dose group showing the most significant protective effect on disc structure and matrix.The protein and mRNA expression lev-els of HIF-1α and p16 were significantly decreased(P＜0.05 or P＜0.01),and the protein and mRNA expression levels of SIRT1 were significantly increased(P＜0.05 or P＜0.01).There were no statistically significant differences in the protein and mRNA expression levels of Ki-67(P＞0.05).Conclusion:Zhuangyao Tongluo formula can inhibit the senes-cence of rat nucleus pulposus cells and ameliorate intervertebral disc tissue injury,thereby playing a role in delaying IVDD.Its mechanism may be related to the regulation of the SIRT1/HIF-1α signaling pathway.