独活寄生汤含药血清对兔退变软骨细胞“caveolin-p38MAPK”信号通路调控作用的影响  

Effects of Serum Medicated with Duhuo Jisheng Decoction on the Induced Degeneration of Articular Chondrocytes in Vitro Culture Based upon “caveolin-p38MAPK” Signal Pathway

作　　者：林飞太[1] 林煜[1] 张怡元[1] 夏春[2] 张兵[3] 肖莉莉[1] 冯尔宥[1] 王武炼[1] 

机构地区：[1]厦门大学附属福州第二医院,福建福州350007 [2]厦门大学附属中山医院,福建厦门361004 [3]厦门大学医学院,福建厦门361005

出　　处：《风湿病与关节炎》2015年第4期5-9,共5页Rheumatism and Arthritis

基　　金：国家自然科学基金资助项目(81302986);福建省卫生厅青年科研课题资助计划项目(2012-2-69);福建省自然科学基金(2013J01389);福州市卫生系统科技计划(2013-S-wq6)

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 m RNA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 m RNA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。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 path- way 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 carti- lage 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 chon- drocytes.The degenerated cartilage cell models in vitro were established,which were intervened by the serum medicated with Duhuo Jisheng Decoction （ drug containing serum group ） and the blank serum （ blank serumgroup ） after 36 h,collecting chondrocytes The Western Blot method was used to detect the expression of degenerated cartilage cells caveolin-1 and p-p38 protein after serum intervention.The RT-PCR method was usedto detect the expression of degenerated cartilage cells IL-113,TNF-a,MMP-3,MMP-13 and caveolin-1 mRNA after serum intervention.Results：The concentration of 15% and 36 h sampling time point serum on the prolifera- tion;the activation of ＂caveolin-p38MAPK＂ signaling pathway in the degeneration of cartilage cell,Duhuo Jisheng decoction containing serum can inhibit the expression of caveolin-1,p-p38 protein expression and IL-1β,TNF-α, MMP-3,MMP-13,caveolin-l,mRNA,the difference was statistically significant （P 〈 0.05 ） .Conclusion：Duhuo Jisheng decoction can inhibit the ＂caveolin-p38MAPK＂ signal pathway and its downstream effector molecules, which can effectively inhibit the apoptosis of cartilage cells.

关 键 词：骨关节炎 独活寄生汤 软骨细胞 细胞凋亡 信号通路 

分 类 号：R285.5[医药卫生—中药学]