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目的：探讨独活-桑寄生组分通过激活PINK1/Parkin及Nrf2通路改善机械压力诱导SD大鼠椎间盘退变的作用机制。方法：检索独活-桑寄生药对核心组分，将SD大鼠随机分为正常对照组、假手术对照组、模型对照组、槲皮素50 mg/kg+蛇床子素20 mg/kg+二氢欧山芹醇当归酸酯20 mg/kg组、槲皮素100 mg/kg+蛇床子素40 mg/kg+二氢欧山芹醇当归酸酯40 mg/kg组及线粒体靶向抗氧化剂依拉米肽3 mg/kg组，每组10只。采用大鼠尾部椎间盘弯折加压法构建椎间盘退变模型，并通过X线评估模型建立情况；利用MRI、HE染色及番红O-固绿染色观察椎间盘退变进程；采用TUNEL染色检测髓核组织凋亡水平；DCFH-DA荧光探针和ELISA法评估氧化应激水平；Western blot法检测PINK1/Parkin通路和Nrf2通路相关蛋白表达。结果：与正常对照组相比，模型对照组大鼠椎间盘MRI、HE及番红O-固绿染色呈现典型椎间盘退变特征，TUNEL染色细胞凋亡率显著增高(P<0.01)，DCFH-DA荧光探针检测ROS水平显著升高(P<0.01)，ELISA检测MDA及蛋白质羰基含量显著增加(P<0.01)、T-SOD及GSH-PX含量显著减少(P<0.01)，Western blot检测BAX、caspase 3、cleaved-caspase 3、cytochrome c、Beclin1、LC3II/I、Parkin、PINK1、P62、Keap1蛋白表达均显著上调(P<0.05或P<0.01)，BCL-2及Nrf2蛋白表达均显著下调(P<0.05或P<0.01)；与模型对照组相比，中药组分各组及线粒体靶向抗氧化剂依拉米肽3 mg/kg组椎间盘退变呈不同程度改善，细胞凋亡率及ROS水平高度显著降低(P<0.01)，MDA及蛋白质羰基含量降低(P<0.05或P<0.01)、T-SOD及GSH-PX升高(P<0.05)，BAX、cleaved-caspase 3、cytochrome c、P62、Keap1均下调(P<0.05或P<0.01)、BCL-2、Beclin1、LC3II/I、Parkin、PINK1、Nrf2均上调(P<0.05或P<0.01)。其中，槲皮素100 mg/kg+蛇床子素40 mg/kg+二氢欧山芹醇当归酸酯40 mg/kg组干预效果更为显著。结论：“祛止补益”中药组分可协同调控PINK1/Parkin与Nrf2信号通路，抑制压力诱导的髓核细胞凋亡，从而延缓椎间盘退变进展。
Abstract：Objective: To investigate the mechanisms through which Duhuo (独活)-Sangjisheng (桑寄生) components ameliorate compression-induced intervertebral disc degeneration (IVDD) in SD rats through activating the PTEN-induced kinase 1 (PINK1)/Parkin and nuclear factor erythroid 2-related factor 2 (Nrf2) pathways. Methods: The core components in the herb pair Duhuo-Sangjisheng were identified. SD rats were randomized into a normal control group， a sham group， a model control group， a quercetin (50 mg/kg) + osthole (20 mg/kg) + columbianadin (20 mg/kg) group， a quercetin (100 mg/kg) + osthole (40 mg/kg) + columbianadin (40 mg/kg) group， and a mitochondrion-targeted antioxidant (3 mg/kg) group， with 10 rats in each group. A rat model of IVDD was established by tail intervertebral disc bending and compression. X-ray imaging was employed to evaluate the modeling results. Magnetic resonance imaging (MRI)， hematoxylin-eosin (HE) staining， and Safranin O-Fast Green staining were employed to observe the degree of intervertebral disc degeneration. Terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay was employed to assess apoptosis levels in the nucleus pulposus (NP) tissue. The DCFH-DA fluorescence probe and enzyme-linked immunosorbent assay (ELISA) were utilized to evaluate oxidative stress levels. Western blot analysis was performed to determine the expression levels of proteins related to the PINK1/Parkin and Nrf2 pathways. Results: Compared with the normal control group， the model control group exhibited typical IVDD features， increased apoptotic rate (P<0.01)， elevated reactive oxygen species (ROS)， malondialdehyde (MDA)， and protein carbonyl levels (P<0.01)， declined levels of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-PX) (P<0.01)， up-regulated protein levels of BCL-2-associated X protein (BAX)， cysteinyl aspartate-specific proteinase 3 (Caspase 3) ， Cleaved-caspase 3， Cytochrome c， Beclin1， microtubule-associated protein 1 light chain 3 subunit II/I (LC3II/I)， Parkin， PINK1， P62， and Kelch-like ECH-associated protein 1 (Keap1) (P<0.05 or P<0.01)， and down-regulated protein levels of B-cell lymphoma 2 (BCL-2) and Nrf2 (P<0.05 or P<0.01). Compared with the model control group， the Chinese medicine component groups and the mitochondrion-targeted antioxidant elamipretide (3 mg/kg) group exhibited varying degrees of alleviation in IVDD， decreased apoptotic rate and ROS levels (P<0.01)， lowered MDA and protein carbonyl levels (P<0.05 or P<0.01)， elevated T-SOD and GSH-PX levels (P<0.05)， down-regulated protein levels of BAX， Cleaved-caspase 3， Cytochrome c， P62， and Keap1 (P<0.05 or P<0.01)， and up-regulated protein levels of BCL-2， Beclin1， LC3II/I， Parkin， PINK1， and Nrf2 (P<0.05 or P<0.01). Among them， the quercetin (100 mg/kg) + osthole (40 mg/kg) + columbianadin (40 mg/kg) group showed the most pronounced therapeutic effects. Conclusion: The Chinese medicine components with the effect of Quzhibuyi (祛止补益) can synergistically modulate both PINK1/Parkin and Nrf2 pathways to inhibit compression-induced apoptosis in NP， thereby delaying the progression of IVDD.
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基本信息:
DOI：10.13412/j.cnki.zyyl.20260716.001
中图分类号:R285.5
引用信息:
[1]冯超群,赵敏,罗茂银,等.基于“祛止补益”理论探究独活-桑寄生组分蛇床子素、二氢欧山芹醇当归酸酯、槲皮素配伍调控PINK1/Parkin和Nrf2通路改善机械应力诱导大鼠椎间盘退变的机制研究[J].中药药理与临床().DOI:10.13412/j.cnki.zyyl.20260716.001.
基金信息:
四川省科技厅（编号：2026NSFSC1870）; 四川省干保课题（川健研2025-501）; 四川省中医药管理局（编号：2024MS551）
2026-07-16
2026-07-16
2026-07-16