盘龙七片调控SIRT1/NF-κB通路对骨关节炎软骨细胞凋亡的影响

Effect of Panlongqi Tablet on Chondrocyte Apoptosis in Osteoarthritis by Regulating the Silent Information Regulator Type 1/Nuclear Factor-kappa B Pathway

目的观察盘龙七片对骨关节炎(osteoarthritis,OA)小鼠沉默信息转录调控因子(silent information regulator type 1,SIRT1)/核因子-κB(nuclear factor-kappa B,NF-κB)通路及软骨细胞凋亡的影响。方法手术切除右膝关节内侧半月板和前交叉韧带复制小鼠OA模型。将小鼠分为正常对照组,模型组,阳性对照组,盘龙七片低、中、高剂量组。通过番红O-快速绿染色观察膝关节结构变化,进行Mankin评分评估关节炎严重程度,流式细胞术检测软骨细胞凋亡指数,qRT-PCR检测Bax、Bcl-2、Caspase-3 mRNA表达水平,Western blot法检测软骨细胞SIRT1、NF-κB蛋白表达水平。结果与正常对照组比较,模型组小鼠膝关节软骨细胞凋亡率增加,Bax、Caspase-3 mRNA和NF-κB蛋白表达水平显著增加,Bcl-2 mRNA和SIRT1蛋白表达水平显著降低,差异均有统计学意义(P<0.05);与模型组比较,阳性对照组与盘龙七片低、中、高剂量组小鼠膝关节软骨细胞凋亡率显著降低,Bax、Caspase-3 mRNA和NF-κB蛋白表达水平显著降低,Bcl-2 mRNA和SIRT1蛋白表达水平显著增加,差异均有统计学意义(P<0.05),盘龙七片的作用呈明显的剂量依赖性。结论盘龙七片可抑制OA小鼠膝关节软骨细胞凋亡,其作用机制可能与调节SIRT1/NF-κB信号通路相关。

Objective To investigate the effect of Panlongqi Tablet on the silent information regulator type 1(SIRT1)/nuclear factor-kappa B(NF-κB)pathway and chondrocyte apoptosis in osteoarthritis(OA)mice.Methods A mouse model of OA was established by surgical resection of the medial meniscus and the anterior cruciate ligament of the right knee,and the mice were divided into normal control group,model group,positive control group,and low-,middle-,and high-dose Panlongqi Tablet groups.Safranin O-fast green staining was performed to observe the changes of knee joint structure;Mankin score was used to evaluate the severity of arthritis;flow cytometry was used to measure chondrocyte apoptosis index;qRT-PCR was used to measure the mRNA expression levels of Bax,Bcl-2,and caspase-3,and Western blot was used to measure the protein expression levels of SIRT1 and NF-κB.Results Compared with the normal control group,the model group had significant increases in the apoptosis rate of knee articular chondrocytes,the mRNA expression levels of Bax and caspase-3,and the protein expression level of NF-κB,as well as significant reductions in the mRNA expression level of Bcl-2 and the protein expression level of SIRT1(P<0.05).Compared with the model group,the positive control group and the low-,middle-,and high-dose Panlongqi Tablet groups had significant reductions in the apoptosis rate of knee articular chondrocytes,the mRNA expression levels of Bax and caspase-3,and the protein expression level of NF-κB,as well as significant increases in the mRNA expression level of Bcl-2 and the protein expression level of SIRT1(P<0.05),and Panlongqi Tablet exerted an effect in a dose-dependent manner.Conclusion Panlongqi Tablet can inhibit the apoptosis of knee articular chondrocytes in OA mice,possibly by regulating the SIRT1/NF-κB signaling pathway.