白芍总苷对人骨关节炎软骨细胞增殖及p38MAPK蛋白表达的影响

Effects of total glucosides of paeonia on proliferation of osteoarthritic chondrocytes by regulating p38MAPK

目的:研究白芍总苷对人骨关节炎软骨细胞增殖和p38MAPK表达的影响。方法:分离膝骨关节炎患者的关节软骨细胞,分为4组,对照组只加培养基,白芍总苷组加入白芍总苷终浓度为8μmol/L的培养基,激动剂组加入p38MAPK信号通路激动剂anisomycin终浓度为2μg/L的培养基,联合组加入含白芍总苷和anisomycin的培养基(终浓度同前)。分组培养24、48、72 h后,MTT法检测细胞增殖能力;分组培养48 h后,碘化丙啶单染法检测细胞周期,qRT-PCR法检测细胞中Ki67、PCNA mRNA表达水平,Western blot法检测细胞中p38MAPK和磷酸化p38MAPK(p-p38MAPK)蛋白表达水平。结果:Anisomycin单独作用可抑制骨关节炎软骨细胞的增殖,阻滞细胞周期进展,促进p38MAPK和p-p38MAPK蛋白的表达(P <0. 05);白芍总苷单独作用则促进细胞增殖和细胞周期进展,抑制p38MAPK和p-p38MAPK蛋白的表达(P <0. 05);两者联用,白芍总苷可以拮抗anisomycin诱导的骨关节炎软骨细胞p38MAPK和p-p38MAPK的表达增加和增殖抑制作用(P <0. 05)。结论:白芍总苷可能通过抑制p38MAPK信号通路,促进人骨关节炎软骨细胞的增殖。

Aim:To study the effects of total glucosides of paeonia(TGP)on the proliferation and p38MAPK protein expression of human osteoarthritic chondrocytes.Methods:Human osteoarthritic chondrocytes were isolated from the patients with knee osteoarthritis,and were allocated into 4 groups,TGP group(cultured with 8μmol/L TGP),agonist group(cultured with 2μg/L anisomycin which was p38MAPK signal pathway agonist),combination group(cultured with 8μmol/L TGP and 2μg/L anisomycin),blank control group(cultured without TGP or anisomycin).After 24,48,72 hour culture,cell proliferation was detected by MTT assay.After 48 hour culture,the cell cycle was detected by PI monochrome staining,the expressions of Ki67 and PCNA mRNA was detected by qRT-PCR,and the p38MAPK and phosphorylated-p38MAPK(p-p38MAPK)proteins were detected by Western blot.Results:When treated with anisomycin,the proliferation ability of human osteoarthritic chondrocytes was decreased,the cell cycle was blocked at G0/G1 phase,and the expression levels of p38MAPK and p-p38MAPK were increased(P<0.05).When treated with TGP,the proliferation ability of human osteoarthritic chondrocytes was increased,the cell cycle was progressing,and the expression levels of p38MAPK and p-p38MAPK were decreased(P<0.05).The combination of the two drugs antagonized each other(P<0.05).Conclusion:TGP may promote the proliferation of human osteoarthritic chondrocytes by inhibiting p38MAPK signaling pathway.