文献类型：期刊文献

中文题名：当归-白芍联合BM-MSCs移植对NASH相关肝硬化小鼠肝脏炎症及肝细胞再生的影响

英文题名：Effects of Angelicae Sinensis Radix-Paeoniae Radix alba Combined with BM-MSCs Transplantation on Liver Inflammation and Hepatocytes Regeneration in Mice with NASH Related Cirrhosis

作者：姚凝[1];王钰涵[2];王昕[3];杨芳丽[4];王馨[1];左小宁[2];秦英[2];夏艳青[5];韩金孝[6];田利民[7]

第一作者：姚凝

机构：[1]甘肃中医药大学第一临床医学院,甘肃兰州730000;[2]苏州大学苏州医学院,江苏苏州215123;[3]甘肃中医药大学中西医结合学院,甘肃兰州730000;[4]青海省交通医院,青海西宁810008;[5]兰州市第一人民医院,甘肃兰州730050;[6]金川集团职工医院,甘肃金昌737100;[7]四川省医学科学院·四川省人民医院,电子科技大学医学院,四川成都610072

第一机构：甘肃中医药大学临床医学院

年份：2024

卷号：31

期号：11

起止页码：129

中文期刊名：中国中医药信息杂志

外文期刊名：Chinese Journal of Information on Traditional Chinese Medicine

收录：CSTPCD;;CSCD:【CSCD_E2023_2024】；

基金：国家自然科学基金(82160845);甘肃省自然科学基金(20JR5RA181);甘肃省教育厅高等学校创新基金(2021B-173);甘肃省兰州市城关区科技计划项目(2021-2-5);敦煌医学与转化教育部重点实验室基金项目(DHYX20-11);甘肃中医药大学引进人才科研启动基金项目(2024YJRC-01)。

语种：中文

中文关键词：当归-白芍;骨髓间充质干细胞;非酒精性脂肪性肝炎;肝硬化;TLR9信号通路;小鼠

外文关键词：Angelicae Sinensis Radix-Paeoniae Radix alba;BM-MSCs;NASH;cirrhosis;TLR9 signaling pathway;mice

摘要：目的观察当归-白芍药对联合骨髓间充质干细胞(BM-MSCs)移植对非酒精性脂肪性肝炎(NASH)相关肝硬化小鼠肝脏炎症及肝细胞再生的影响,探讨其可能机制。方法采用西式饮食联合四氯化碳复合诱导法制备NASH相关肝硬化小鼠模型,将小鼠随机分为对照组、模型组、当归-白芍组、BM-MSCs组和当归-白芍+BM-MSCs组,每组12只。造模成功后,按分组予相应干预,连续4周。HE染色观察肝组织形态;检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆红素(TBil)、三酰甘油(TG)、白蛋白(ALB)、甲胎蛋白(AFP)含量及肝组织白细胞介素-1β(IL-1β)、肿瘤坏死因子(TNF)-α、肝细胞生长因子(HGF)含量;RT-qPCR检测肝细胞Toll样受体9(TLR9)、髓样分化因子88(MyD88)、TNF受体相关因子6(TRAF6)、核因子-κB(NF-κB)p65 mRNA表达;分离肝脏原代细胞,行体外CpG ODN 2216诱导刺激,提取细胞上清液,检测IL-1β和TNF-α含量。结果与对照组比较,模型组小鼠肝组织炎性细胞浸润,伴肝细胞坏死和胶原沉积,形成假小叶;血清ALT、AST、TBil、TG含量升高,ALB含量下降,肝组织HGF含量下降,IL-1β、TNF-α含量升高,差异均有统计学意义(P<0.05);肝细胞TLR9、MyD88、TRAF6、NF-κBp65 mRNA表达升高(P<0.05),CpG ODN 2216诱导后肝细胞上清液IL-1β、TNF-α含量升高(P<0.05)。与模型组比较,当归-白芍+BM-MSCs组小鼠肝组织炎性细胞浸润及肝细胞坏死减少,肝纤维化程度减轻,当归-白芍组和BM-MSCs组小鼠肝组织损伤轻度好转;除BM-MSCs组血清TG含量外,各干预组检测指标均明显改善(P<0.05)。与当归-白芍组及BM-MSCs组比较,当归-白芍+BM-MSCs组相关检测指标差异均有统计学意义(P<0.05)。结论当归-白芍联合BM-MSCs移植可有效改善NASH相关肝硬化小鼠肝功能,促进肝细胞再生,其机制与下调TLR9/NF-κB信号通路表达及功能,抑制炎症因子分泌,改善肝脏炎症微环境有关。
Objective To observe the effects of Angelicae Sinensis Radix-Paeoniae Radix alba combined with bone marrow mesenchymal stem cells(BM-MSCs)transplantation on liver inflammation and hepatocytes regeneration in non-alcoholic steatohepatitis(NASH)associated cirrhosis;To discuss its possible mechanism.Methods West diet combined with carbon tetrachloride was used to prepare the NASH related cirrhosis model.The mice were randomly divided into control group,model group,Angelicae Sinensis Radix-Paeoniae Radix alba group,BM-MSCs group,and Angelicae Sinensis Radix-Paeoniae Radix alba+BM-MSCs group,with 12 mice in each group.After successful modeling,corresponding interventions were given according to grouping for 4 consecutive weeks.HE staining was used to observe the morphology of liver tissue;the contents of serum ALT,AST,TBil,TG,ALB,AFP,as well as the contents of IL-1β,TNF-αand HGF in liver tissue were detected;RT-qPCR was used to detect TLR9,MyD88,TRAF6 and NF-κBp65 mRNA expression in liver cells;primary liver cells were separated,CpG ODN 2216 stimulation was induced in vitro,cells supernatant was extracted,and IL-1βand TNF-αcontent were detected.Results Compared with the control group,inflammatory cell infiltrated in liver tissue of model group mice,accompanied by hepatocyte necrosis and collagen deposition,forming pseudo lobules;the contents of serum ALT,AST,TBil and TG increased,the content of ALB decreased,the content of HGF in liver tissue decreased,the contents of IL-1βand TNF-αincreased,with statistical significance(P<0.05);the mRNA expressions of TLR9,MyD88,TRAF6 and NF-κBp65 in liver cells increased(P<0.05),and the contents of IL-1βand TNF-αin cells supernatant increased after CpG ODN 2216 induction(P<0.05).Compared with the model group,the inflammatory infiltration in liver tissue and necrosis of liver cells were reduced and the degree of liver fibrosis was alleviated in the Angelicae Sinensis Radix-Paeoniae Radix alba+BM-MSCs group,the liver tissue damage in Angelicae Sinensis Radix-Paeoniae Radix alba group and BM-MSCs group were slightly improved;except for the TG content in BM-MSCs group,all detection indicators showed significant improvement in each intervention group(P<0.05).Compared with the Angelicae Sinensis Radix-Paeoniae Radix alba group and BM-MSCs group,the Angelicae Sinensis Radix-Paeoniae Radix alba+BM-MSCs group showed statistical significance in related detection indicators(P<0.05),and demonstrated a synergistic effect.Conclusion Angelicae Sinensis Radix-Paeoniae Radix alba combined with BM-MSCs transplantation could effectively improve the liver function and promote liver cell regeneration.The mechanism is associated with the down-regulation of TLR9/NF-κB signaling pathway expressions and function,the inhibition of inflammatory cytokines secretions,and the improvement of liver inflammatory microenviroment.