脐带间充质干细胞介导TLR4/NF-κB炎症通路及氧化应激改善NAFLD大鼠肝脏损伤作用研究  

Human umbilical cord mesenchymal stem cells alleviate liver damage of non-alcoholic fatty liver disease rats by inhibitting TLR4/NF-κB and oxidative stress

作　　者：王文清[1] 易文城[1] 林拥华[1] Wang Wenqing;Yi Wencheng;Lin Yonghua(Department of General Surgery,the Second Affiliated Hospital of Fujian Medical University,Quanzhou 362000,China)

机构地区：[1]福建医科大学附属第二医院普通外科,泉州362000

出　　处：《中华细胞与干细胞杂志（电子版）》2018年第5期297-302,共6页Chinese Journal of Cell and Stem Cell（Electronic Edition）

摘　　要：目的探讨脐带间充质干细胞(hUC-MSCs)对非酒精性脂肪肝病(NAFLD)大鼠肝脏损伤的作用及其与TLR4/NF-κB炎症通路和氧化应激的关系。方法 SD大鼠随机分为3组:正常对照组(NC组)、模型对照组(MC组)和hUC-MSCs治疗组(MC+hUC-MSCs组)。每组大鼠均为10只,分别喂食常规或高糖高脂饲料并给予相应治疗8周,其中MC+hUC-MSCs组每两周尾静脉注射含5×106个hUC-MSCs。检测各组肝指数、谷丙转氨酶(ALT)、谷草转氨酶(AST)、甘油三酯(TG)、胆固醇(TC)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)和胰岛素抵抗指数(HOMA-IR)。光镜和电镜观察大鼠肝脏组织病理改变,评估NAFLD活动度积分(NAS);Western Blot法检测大鼠肝脏组织超氧化物歧化酶(SOD)、8-羟基脱氧鸟苷(8-OHdG)、肿瘤坏死因子(TNF-α)、白介素-6(IL-6)Toll样受体4(TLR4)和核因子-κB (NF-κB)蛋白表达。组间比较采用单因素方差分析。结果干预结束后,(1)NC组TG(0.96±0.29) mmol/L、TC(2.23±0.37)mmol/L、LDL(0.71±0.18)mmol/L、HDL(2.95±0.27)mmol/L,与MC组TG(5.79±0.68)mmol/L、TC(6.08±0.79)mmol/L、LDL(6.06±0.31)mmol/L、HDL(0.75±0.18) mmol/L比较差异具有统计学意义(均P <0.05);与MC组比较,MC+hUC-MSCs组上述指标差异具有统计学意义(均P <0.05)。(2)光镜下NC组肝细胞形态正常;MC组出现肝细胞脂肪变性、肝小叶排列不齐及炎症细胞浸润;而MC+hUC-MSCs组肝组织病理改变明显好转。与NC组比较,MC组肝组织NAS升高;与MC组比较,MC+hUC-MSCs组NAS降低[(0.36±0.16)分vs(8.72±0.35)分、(4.78±0.51)分,P <0.05]。电镜下亦观察到与MC组比较,MC+hUC-MSCs大鼠肝细胞脂肪变性、胞核变形、内质网和线粒体形态异常明显改善。(3)与NC组比较,MC组大鼠TNF-α、IL-6、8-OHdG、TLR4和NF-κB蛋白表达均增高(均P <0.05),SOD降低(P <0.05);与MC组比较,MC+hUC-MSCs组大鼠上述指标改善(P <0.05)。结论 hUC-MSCs通过抑制氧化应激和TLR4/NF-κB而保护NAFLD大鼠肝脏功能。Objective To evaluate the effect of human umbilical cord mesenchymal stem cells on liver injury and TLR4/NF-κB and oxidative stress of non-alcoholic fatty liver disease rats.Methods SD rats were randomly divided into three groups:control group(NC),non-alcoholic fatty liver disease model group(MC)and hUC-MSCs treatment group(MC+hUC-MSCs)(n=10 for each group).The rats in the control group were fed with conventional feed,and the rats in MC and MC+hUC-MSCs group were fed with high glucose and high fat diet.Once every two weeks,the rats in hUC-MSCs group were injected 5×10^6 hUC-MSCs by caudal vein.At the end of therapy,liver index,ALT,AST,TG,TC,LDL,HDL and HOMA-IR were measured.Liver tissues were observed with light microscope and transmission electronic microscope.The protein expressions of 8-OhdG,SOD,TNF-α,IL-6,TLR4 and NF-κB of liver tissues were detected by Western Blot.Results(1)At the end of therapy,parameters in the control group were as follow:TG(0.96±0.29)mmol/L,TC(2.23±0.37)mmol/L,LDL(0.71±0.18)mmol/L,HDL(2.95±0.27)mmol/L;for the MC group,they were:TG(5.79±0.68)mmol/L,TC(6.08±0.79)mmol/L,LDL(6.06±0.31)mmol/L,HDL(0.75±0.18)mmol/L compared with the control group,the value of the liver index,ALT,AST,TG,TC,LDL and HOMA-IR in the MC group were significantly increased(P<0.05),and HDL decreased(P<0.05),which were reversed in the MC+hUC-MSCs group(P<0.05).(2)HE staining showed fatty degeneration,disorder and inflammatory cell infiltration in the MC group.However,the above liver histopathology changes were improved in the MC+hUC-MSCs group.Compared with the control group,the value of NAS in the MC group significantly incresed.Compared with the MC group,the value of NAS was significantly declined in the MC+hUC-MSCs groups(0.36±0.16)vs(8.72±0.35)、(4.78±0.51),P<0.05.Transmission electron microscope showed hepatocyte steatosis,cell nucleus deformation,endoplasmic reticulum and mitochondrial abnormalities were significantly improved in the MC+hUC-MSCs group.(3)Compared with the control group,the protein

关 键 词：非酒精性脂肪肝 TLR4 NF-ΚB 氧化应激 脐带间充质干细胞 

分 类 号：R575[医药卫生—消化系统]