摘要
目的探讨骨髓间充质干细胞(MSCs)移植对大鼠肝纤维化模型的作用与机制。方法分离大鼠MSCs培养传代至第4代。采用大鼠腹腔注射四氯化碳制造肝纤维模型。将造模成功SD大鼠60只,随机分为3组,每组20只:(1)对照组:鼠尾静脉注射等量的生理盐水;(2)MSCs组:鼠尾静脉注射MSCs悬液;(3)诱导组:鼠尾静脉注射经肝细胞生长因子(HGF)诱导14 d后的MSCs悬液。于移植后第1周、2周、3周、4周分别处死大鼠5只,检测大鼠血清透明质酸(HA)、层黏蛋白(LN)、Ⅳ型胶原水平;光镜下观察肝组织纤维化程度;分别用PCR法及Western检测大鼠肝脏Col-Ⅰ、RhoA、Cdc42、Rac1 mRNA基因及其蛋白质表达水平。结果(1)诱导组与MSCs组的纤维化程度评分随着时间延长逐渐下降,且在第4周诱导组评分明显低于MSCs组(P<0.05)。(2)移植3周后各组血清HA、LN、Ⅳ型胶原含量均显著下降,4周时诱导组和MSCs组各指标含量明显低于对照组,并且诱导组低于MSCs组(P<0.05)。(3)MSCs移植后诱导组、MSCs组大鼠肝脏组织Col-Ⅰ、RhoA、Cdc42、Rac1 mRNA和蛋白表达均随时间延长而下降,移植4周时诱导组各项指标明显低于MSCs组和对照组(P<0.05)。结论 MSCs移植可抑制肝纤维化大鼠肝组织中HA、LN、Ⅳ型胶原的分泌,并可下调肝纤维化大鼠肝组织Rho信号通路相关因子RhoA,Cdc42,Rac1 mRNA及蛋白表达。HGF诱导MSCs后抗大鼠肝纤维化作用显著增强。
Objective To study the effect of mesenchymal stem cells （ MSCs ） transplantation on rats with hepatic fibrosis and its mechanism.Methods MSCs was isolated from Sprague-Dawley（SD） rats and cultured for 4 passages.The hepatic fibrosis model of SD rats was induced by subcutaneous injection of CCl 4.Sixty SD hepatic fibrosis rats were divided into 3 groups randomly （20 rats in each group）：①Control group：The rats were treated by infusion of normal saline via tail vein.②MSCs group：The rats were treated by injection of MSCs suspension via tail vein .③Induced group：The rats were treated by injection of hepatocyte growth factor （HGF）-induced MSCs suspension via tail vein 14 days after induction.The rats were killed 1 week,2 weeks,3 weeks,4 weeks after MSCs transplantation,respectively,5 rats for each batch.The serum levels of hyaluronic acid （HA）,laminin（LN）,collagen Ⅳ（Col-Ⅳ） were detected.The degree of hepatic fibrosis was observed with light microscope.The mRNA expressions of collagenⅠ（Col-Ⅰ）,RhoA,Cdc42,ras-related C3 botulinum toxin substrate（Rac1） in hepatic tissues were detected by PCR ,whereas the protein expressions of Col-Ⅰ,RhoA,Cdc42,Rac1 in hepatic tissues were detected by Western blot method .Results ①The score of hepatic fibrosis degree was smaller with increasing time in the induced group and MSCs group ,which was smaller in the induced group in the fourth week in contrast with that in the MSCs group （P〈0.05）.②The serum levels of HA,LN,Col-Ⅳdecreased significantly in eachgroup 3 weeks after transplantation ,and they were significantly lower in the induced group ,MSCs group compared with control group ,which were lower in the induced group in contrast with those in the MSCs group （ P〈0 .05 ） .③After MSCs transplantation,the mRNA and protein expressions of Col-Ⅰ,RhoA,Cdc42,Rac1 in hepatic tissues reduced more signif-icantly in the MSCs group and induced group with increasing time ,which were significantly lower in the induced group 4 weeks after transplantation compared with MSCs group and control group （P〈0.05）.Conclusion MSCs transplantation may suppress the secretion of HA ,LN, Col-Ⅳin the tissues of hepatic fibrosis rats as well as down-regulate the mRNA and protein expressions of Col-Ⅰ,RhoA,Cdc42,Rac1,which were Rho signal channel-related factors,in the tissues of hepatic fibrosis rats.HGF-induced MSCs can further strengthen this effect .
出处
《广西医学》
CAS
2014年第2期141-145,共5页
Guangxi Medical Journal
基金
国家自然科学基金资助项目(No.81260083
31360221)
广西自然科学基金资助项目(2012GXNSFAA053140)
