摘要
目的:探究三七总皂苷(PNS)对骨关节炎(OA)大鼠软骨细胞焦亡的影响及其作用机制。方法:50只SD大鼠随机分为假手术组(Sham)、模型组(Model)、三七总皂苷低剂量组(PNS-L,50 mg/kg)、三七总皂苷高剂量组(PNS-H,200 mg/kg)和阳性对照药塞来昔布组(Celecoxib,24 mg/kg),每组各10只。除Sham组,其余各组大鼠均采用Hulth法构建OA模型。药物干预8周后,苏木素-伊红(HE)染色、番红O(SO)染色和阿尔新蓝(Alcian blue)染色观察膝关节组织病理学变化;电子压痛仪和足底热测痛仪检测大鼠压痛阈值、热痛阈值;TUNEL检测软骨细胞焦亡;免疫组化和Western blot检测软骨细胞TLR4/NLRP3/Caspase-1信号通路相关蛋白表达。结果:与Sham组相比,Model组大鼠关节软骨组织坏死,纤维组织增生明显,各层细胞排列紊乱,Mankin’s评分显著增加,有典型的OA病理损伤,大鼠压痛阈值及热痛阈值均显著降低(P<0.01),软骨细胞焦亡率、TLR4阳性细胞率、NLRP3阳性细胞率、Caspase-1阳性细胞率以及软骨组织中TLR4、NLRP3、Caspase-1、IL-1β、IL-18蛋白表达均显著增加(P<0.01)。与Model组相比,PNS各剂量组大鼠关节软骨细胞增多,各层细胞排列相对规则,Mankin’s评分显著降低,病理损伤明显减轻,大鼠压痛阈值及热痛阈值均显著增加(P<0.01),软骨细胞焦亡率、TLR4阳性细胞率、NLRP3阳性细胞率、Caspase-1阳性细胞率以及软骨组织中TLR4、NLRP3、Caspase-1、IL-1β、IL-18蛋白表达均显著降低(P<0.05或P<0.01)。结论:PNS能够抑制OA大鼠软骨细胞焦亡,改善骨关节损伤,其机制可能与抑制TLR4/NLRP3/Caspase-1信号通路相关。
Objective:To investigate the effect and mechanism of Panax notoginseng saponins(PNS)on chondrocyte pyroptosis in osteoarthritis(OA)rats.Methods:50 SD rats were randomly divided into the sham operation group,the model group,the low-dose group of Panax notoginseng saponins(PNS-L,50 mg/kg),and the high-dose group of Panax notoginseng saponins(PNS-H,200 mg/kg),and the positive control drug group of Celecoxib(Celecoxib,24 mg/kg);with 10 rats in each group.Except for the sham operation group,the other groups were all used Hulth method to establish OA model.After 8 weeks of interventions,H staining,SO staining and Alcian blue staining were used to observe histopathological changes of knee joints.Electronic tenderness instrument and plantar thermal analgesia were used to detect tenderness threshold and thermal pain threshold.TUNEL was used to detect chondrocyte pyroptosis.Immunohistochemistry and Western blot were used to detect the expressions of TLR4/NLRP3/Caspase-1 signaling pathway-related proteins in chondrocytes.Results:Compared with those in the sham operation group,the articular cartilage tissue was necrotic,the hyperplasia of fibrous tissue was obvious,the cells in each layer were disordered,the Mankin’s score was significantly increased,there were typical pathological lesions of OA,and the tenderness threshold and thermal pain threshold were significantly decreased(P<0.01);the rate of chondrocyte pyroptosis,TLR4 positive cell rate,NLRP3 positive cell rate,Caspase-1 positive cell rate and the protein expressions of TLR4,NLRP3,Caspase-1,IL-1β and IL-18 in cartilage tissues significantly increased in the model group(P<0.01).Compared with those in the model group,the articular cartilage cells increased,the cells in each layer were relatively regular,the Mankin’s score was significantly reduced,the pathological damage was significantly reduced,and the tenderness threshold and the thermal pain threshold were significantly increased(P<0.01);chondrocyte pyroptosis rate,TLR4 positive cell rate,NLRP3 positive cell rate,Caspase-1 positive cell rate and the protein expressions of TLR4,NLRP3,Caspase-1,IL-1β and IL-18 in cartilage tissues significantly decreased in the PNS groups(P<0.05,P<0.01).Conclusion:PNS can inhibit chondrocyte pyroptosis and improve bone and joint injuries in OA rats,and its mechanism may be related to the inhibition of TLR4/NLRP3/Caspase-1 signaling pathway.
作者
蔡猛
张永宁
CAI Meng;ZHANG Yongning(Zhumadian Central Hospital,Zhumadian 463000,China;School of Medicine,Huanghuai College,Zhumadian 463000,China)
出处
《中医药信息》
2023年第2期11-17,共7页
Information on Traditional Chinese Medicine
基金
河南省高等学校重点科研项目(20B320120)。
