苍术内酯Ⅱ对骨关节炎大鼠软骨损伤、血清炎症因子和氧化应激的调节作用及机制研究

目的:研究苍术内酯Ⅱ(atractylenolide Ⅱ,AT-Ⅱ)对骨关节炎(osteoarthritis,OA)大鼠软骨损伤、血清炎症因子和氧化应激的调节作用及机制。方法:将30只SD大鼠随机分为对照组,模型组,AT-Ⅱ低、中、高剂量(5 mg·kg^-1、10 mg·kg^-1、20 mg·kg^-1)组,用5%(w/v)木瓜蛋白酶制备OA模型。各组大鼠用相应药物处理3周后染色观察各组软骨损伤情况,软骨组织中细胞外基质蛋白、细胞凋亡蛋白、NF-κB途径、STAT3途径相关蛋白表达情况,并检测大鼠外周血中氧化应激相关因子及炎症细胞因子的水平。结果:AT-Ⅱ处理OA模型后,能改善软骨损伤情况,且改善程度呈浓度依赖;AT-Ⅱ处理能促进OA模型中Coll-Ⅱ和蛋白多糖(aggrecan)的mRNA和蛋白表达,抑制基质金属蛋白酶13(MMP-13)的mRNA和蛋白表达(P<0.01);AT-Ⅱ处理能抑制细胞凋亡相关蛋白Caspase-3、Caspase-9、Bax的表达,促进Bcl-2蛋白的表达(P<0.01);AT-Ⅱ处理能提高血清中超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽(glutathione,GSH)、白细胞介素-10(interleukin-10,IL-10)水平,降低丙二醛(monochrome display adapter,MDA)、乳酸脱氢酶(lactate dehydrogenase,LDH)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor,TNF-α)水平(P<0.01);AT-Ⅱ处理能够抑制P65和STAT3蛋白的磷酸化(P<0.05)。结论:AT-Ⅱ可能通过抑制NF-κB途径及STAT3途径蛋白磷酸化抑制细胞凋亡,降低氧化应激及炎症反应从而缓解OA模型大鼠软骨损伤。

Effects and Mechanism of Atractylolide II on Cartilage Injury,Serum Inflammatory Factors and Oxidative Stress in Osteoarthritis Rats

Objective:To study the regulatory effect and mechanism of atractylenolide II(AT-II)on cartilage injury,serum inflammatory factors and oxidative stress in osteoarthritis(OA)rats.Methods:30 SD rats were randomly divided into control group,model group,AT-II low,middle,high dosage group(5 mg·kg^-1,10 mg·kg^-1,20 mg·kg^-1).The OA model group was made by 5%(w/V)papain.After 3 weeks of treatment with corresponding drugs,the cartilage injury,the expression of extracellular matrix protein,apoptosis protein,NF-κB pathway and STAT3 pathway related proteins in cartilage tissue were observed,and the levels of oxidative stress related factors and inflammatory cytokines in peripheral blood were detected.Results:After AT-II processes the OA model,it can improve cartilage damage,and the degree of improvement is concentration-dependent;AT-II treatment can promote the mRNA and protein expression of Coll-II and proteoglycan(aggrecan)in the OA model,and inhibit the mRNA and protein expression of matrix metalloproteinase 13(MMP^-13)(P<0.01);AT-II treatment can inhibit the expression of apoptosis-related proteins Caspase-3,Caspase-9,Bax,and promote the expression of Bcl-2 protein(P<0.01);AT-II treatment can increase serum superoxide dismutase(SOD),glutathione(GSH),interleukin^-10(IL^-10)levels,reduce monochrome display adapter(MDA),lactate dehydrogenase(LDH),interleukin^-1β(IL^-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)levels(P<0.01);AT-II treatment can inhibit the phosphorylation of P65 and STAT3 protein(P<0.05).Conclusion:AT-II may inhibit cell apoptosis by inhibiting NF-κB pathway and STAT3 pathway protein phosphorylation,and reduce oxidative stress and inflammation,thereby alleviating cartilage damage in OA rats.