人脐带间充质干细胞来源的细胞外囊泡增强纤维化肝脏再生能力  

Human umbilical cord mesenchymal stem cell-derived extracellular vesicles enhance the regenerative capability of fibrotic liver

作　　者：雷耘果 姚嘉[1] 郑俊[1] 陆桐宇 张杰滨 萧家麒 刘亚松 陈海填 赵雪刚 杨兴业 Lei Yunguo;Yao Jia;Zheng Jun;Lu Tongyu;Zhang Jiebin;Xiao Jiaqi;Liu Yasong;Chen Haitian;Zhao Xuegang;Yang Xingye(Department of Liver Surgery,Liver Transplantation Center,the Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院肝脏外科暨肝移植中心,广州510630

出　　处：《器官移植》2023年第3期379-388,共10页Organ Transplantation

基　　金：广东省自然科学基金面上项目(2019A1515011698)。

摘　　要：目的探讨人脐带间充质干细胞来源的细胞外囊泡(hUC-MSC-EV)在纤维化肝脏再生中的作用。方法将C57BL/6小鼠随机分为正常肝脏70%肝切除组(Oil+PHx组)、肝纤维化70%肝切除组(CCl_(4)+PHx组)、肝纤维化70%肝切除+间充质干细胞来源的细胞外囊泡(MSC-EV)治疗组(CCl_(4)+PHx+MSC-EV组),每组8只。将LX-2细胞分为磷酸盐缓冲液(PBS)组、转化生长因子(TGF)-β组、TGF-β+MSC-EV组。检测各组小鼠肝部分切除术后丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)水平,分析各组小鼠肝组织纤维化及增殖相关指标的表达情况。检测各组LX-2细胞表皮细胞生长因子(EGF)、成纤维母细胞生长因子(FGF)、血管内皮生长因子(VEGF)、肝细胞生长因子(HGF)信使RNA(mRNA)的表达水平。观察对小鼠肝脏HGF表达的影响。结果与Oil+PHx组比较,CCl_(4)+PHx组小鼠血清AST、ALT、LDH水平升高,纤维化程度较高,天狼星红及α-平滑肌肌动蛋白(α-SMA)染色阳性区域面积增大,α-SMA蛋白表达水平升高;与CCl_(4)+PHx组比较,CCl_(4)+PHx+MSC-EV组小鼠血清AST、ALT、LDH水平下降,纤维化程度较轻,天狼星红及α-SMA染色阳性区域面积缩小,α-SMA蛋白表达水平下降,差异均有统计学意义(均为P<0.05)。与Oil+PHx组比较,CCl_(4)+PHx组Ki67、增殖细胞核抗原(PCNA)蛋白表达水平降低;与CCl_(4)+PHx组比较,CCl_(4)+PHx+MSC-EV组Ki67、PCNA蛋白表达水平升高,差异均有统计学意义(均为P<0.05)。与PBS组比较,TGF-β组LX-2细胞内CollagenⅠmRNA表达水平升高,α-SMA蛋白表达水平升高,HGF蛋白表达水平下降;与TGF-β组比较,TGF-β+MSC-EV组LX-2细胞内CollagenⅠmRNA表达水平下降,HGF mRNA和蛋白表达水平升高,α-SMA蛋白表达水平降低,差异均有统计学意义(均为P<0.05)。CCl_(4)+PHx组HGF蛋白表达水平较Oil+PHx组下降,但差异无统计学意义(P>0.05);CCl_(4)+PHx+MSC-EV组HGF蛋白表达水平较CCl_(4)+PHx组上升,差异有统计学意义(PObjective To investigate the role of human umbilical cord mesenchymal stem cell-derived extracellular vesicle(hUC-MSC-EV)in the regeneration of fibrotic liver.Methods C57BL/6 mice were randomly divided into the 70%normal liver resection group(Oil+PHx group),70%liver fibrosis resection group(CCl_(4)+PHx group)and 70%liver fibrosis resection+mesenchymal stem cell-derived extracellular vesicle(MSC-EV)treatment group(CCl_(4)+PHx+MSC-EV group),with 8 mice in each group.LX-2 cell lines were assigned into the phosphate buffer solution(PBS)group,transforming growth factor(TGF)-βgroup and TGF-β+MSC-EV group.The serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and lactate dehydrogenase(LDH)in mice after partial liver resection were detected in each group.The expression levels of liver fibrosis and proliferation-related parameters were analyzed in each group.The messenger RNA(mRNA)expression levels of epidermal growth factor(EGF),fibroblast growth factor(FGF),vascular endothelial growth factor(VEGF)and hepatocyte growth factor(HGF)in LX-2 cells were detected in each group,and their effects on HGF expression in mouse liver were observed.Results Compared with the Oil+PHx group,the serum levels of AST,ALT and LDH were up-regulated,and the degree of fibrosis was more severe,the positive area of Sirius red andα-smooth muscle actin(α-SMA)staining was larger,and the expression level ofα-SMA protein was up-regulated in the CCl_(4)+PHx group.Compared with the CCl_(4)+PHx group,the serum levels of AST,ALT and LDH were decreased,the degree of fibrosis was slighter,the positive area of Sirius red andα-SMA staining was decreased,and the expression level ofα-SMA protein was down-regulated in the CCl_(4)+PHx+MSC-EV group,and the differences were statistically significant(all P<0.05).Compared with the Oil+PHx group,the protein expression levels of Ki67 and proliferating cell nuclear antigen(PCNA)were lower in the CCl_(4)+PHx group.Compared with the CCl_(4)+PHx group,the protein expression levels of Ki67 a

关 键 词：肝纤维化 部分肝切除 肝移植 人脐带间充质干细胞(hUC-MSC) 细胞外囊泡 肝再生 肝细胞生长因子 肝星状细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R575[医药卫生—基础医学]