摘要
目的观察大鼠骨髓间充质干细胞(BMSCs)旁分泌肝细胞生长因子(HGF)对肝星状细胞(HSCs)死亡受体-5(DR5)及caspase-8的影响。方法应用6孔培养板建立双层细胞共培养体系,分A组:对照组;B组:共培养组;C组:TNF-α预处理组;D组:HGF-ShRNA干扰组。ELISA法检测上清液中HGF及TRAIL的浓度;Annexin V-FITC/PI双染法检测细胞凋亡;荧光定量-PCR、Western blot分别检测DR5、caspase-8 mRNA和蛋白的表达。结果 1)TNF-α预处理组中HGF的浓度明显高于BMSCs组及对照组(P<0.01);BMSCs组中TRAIL的浓度明显高于其他3组(P<0.01)。2)TNF-α预处理组中HSCs凋亡率明显高于其他3组且呈时间依赖性(P<0.01);HGF-ShRNA干扰组中HSCs的凋亡率低于共培养组,高于对照组(P<0.01)。3)共培养组及TNF-α预处理组DR5、caspase-8 mRNA及蛋白表达量明显高于对照组及HGF-ShRNA干扰组(P<0.01)。结论 BMSCs旁分泌HGF能上调DR5及caspase-8的表达促进HSCs的凋亡。
Abstract： Objective To observe the effect of HGF from rat bone marrow mesenchymal stem cells paracrine on he- patic stellate cells death Receptor-5 and caspase-8. Methods Six well plates were used to establish co-culture sys- tem. Cells were divided into group A, control group, group B, co-cultured group; group C： TNF-ot pretreatment group; group D： HGF-ShRNA interference group. The concentrations of HGF and TRAIL were measured by ELISA. Apoptosis of HSCs was determined by Annexin-V-FITC/PI; the expression of DR5 mRNA, caspase-8 mR- NA and protein were determined by FQ-PCR and Western blot respectively. Results 1 ） Concentration of HGF in TNF-α pretreatment group was obviously higher than that in BMSCs group and control group （P 〈0. 01 ） ; Concen- tration of TRAIL in the BMSCs group was higher than that in control group and TNF-α pretreament group; 2） Apoptosis rate of HSCs in TNF-α pretreatment group was obviously higher than that of the other three groups andhad time-dependent （P 〈0. 01 ） ; Apoptosis rate of HGF-ShRNA interference group was lower than that of in co- cluture group and higher than that of control group （P 〈 0.05 ）. 3 ） The expressions of DILS, caspase-8 protein in co-culture group and TNF-α pretreatment group were significantly higher than that of in control group and HGF- ShRNA interference group （ P 〈 0. 01 ）. Conclusions BMSCs paracrined HGF can up-regulate the expression of DR5 and caspase-8 which may be associated with the apoptosis of HSCs.
出处
《基础医学与临床》
CSCD
北大核心
2012年第7期804-808,共5页
Basic and Clinical Medicine
基金
广西自然科学基金(0897088)
关键词
骨髓间充质干细胞
肝星状细胞
肿瘤坏死因子相关凋亡诱导配体
肝细胞生长因子
死亡受体-5
bone marrow mesenchymal stem cells
hepatic stellate cells
tumor necrosis factor-related apoptosis-inducing ligand
hepatocyte growth factor
death receptor-5
