独活寄生汤含药血清对兔退变软骨细胞caveolin―p38MAPK信号通路调控作用影响.docVIP

独活寄生汤含药血清对兔退变软骨细胞caveolin―p38MAPK信号通路调控作用影响 　　【摘 要】目的：观察独活寄生汤含药血清对兔退变软骨细胞“caveolin-p38MAPK”信号通路的调控作用，探讨独活寄生汤治疗骨关节炎的作用机制。方法：将20只3月龄新西兰兔随机分为生理盐水组（空白血清组）和独活寄生汤组（含药血清组），每组10只。分别在24 h、36 h、48 h不同采血时间点采集独活寄生汤含药血清和空白血清，将5%、10%、15%、20%不同浓度两种血清作用于体外培养第2代软骨细胞，确定含药血清最佳干预条件；建立体外退变软骨细胞模型，分别给予独活寄生汤含药血清（含药血清组）和空白血清（空白血清组）干预36 h，收集软骨细胞，运用Western Blot法检测血清干预后退变软骨细胞caveolin-1、p38、p-p38蛋白表达，RT-PCR法检测血清干预后退变软骨细胞白细胞介素（IL）-1β、肿瘤坏死因子（TNF）-α、基质金属蛋白酶（MMP）-3、MMP-13、caveolin-1 mRNA表达。结果：浓度为15%的36 h采血时间点含药血清的促增殖作用最明显；退变软骨细胞中存在“caveolin-p38MAPK”信号通路的激活，独活寄生汤含药血清可抑制caveolin-1、p-p38蛋白表达及IL-1β、TNF-α、MMP-3、MMP-13、caveolin-1 mRNA的表达，差异有统计学意义（P 0.05）。结论：独活寄生汤能通过抑制 　　“caveolin-p38MAPK”信号通路的激活及其下游效应分子，从而有效抑制软骨细胞凋亡。 　　【关键词】 骨关节炎；独活寄生汤；软骨细胞；细胞凋亡；信号通路 　　doi：10.3969/j.issn.2095-4174.2015.04.001 　　Effects of Serum Medicated with Duhuo Jisheng Decoction on the Induced Degeneration of Articular Chondrocytes in Vitro Culture Based upon “caveolin-p38MAPK” Signal Pathway 　　LIN Fei-tai，LIN Yu，ZHANG Yi-yuan，XIA Chun，ZHANG Bing，XIAO Li-li，FENG Er-you，WANG Wu-lian 　　【ABSTRACT】Objective：To observe the effects of serum medicated with Duhuo Jisheng Decoction on the induced degeneration of articular chondrocytes in vitro culture based upon “caveolin-p38MAPK” signal pathway and probe the mechanism of Duhuo Jisheng Decoction in the treatment of osteoarthritis.Methods：20 March 　　old New Zealand rabbits were randomly divided into normal saline group（blank serum group）and Duhuo Jisheng Decoction group （drug containing serum group），10 rats in each group.The second generation of cartilage cells was cultured in vitro using different time points of sample collection such as 24 h，36 h and 48 h，serums medicated with 5%，10%，15% and 20% concentrations of Duhuo Jisheng Decoction and blank serums.The best intervention time of medicated serum was determined，which was then used to deal with the degenerating chondrocytes.The degenerated cartilage cell models in vitro were established，which were intervened by the serum medicated