小鼠骨髓间充质干细胞移植对不同剂量X射线所致小鼠肝损伤的干预效应  

Interventional effect of bone marrow mesenchymal stem cell transplantation with different doses of X-ray irradiation induced hepatic injury in mice

作　　者：梁悦 罗蓝 程天宇 陈高峰[1,2] 刘伟[1,2] 慕永平[1,2] 陈佳美[1,2] 刘平[1,2,3] Liang Yue;Luo Lan;Cheng Tianyu;Chen Gaofeng;Liu Wei;Mu Yongping;Chen Jiamei;Liu Ping(Institute of Liver Diseases,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai201203,China;Key Laboratory of Liver and Kidney Diseases(Ministry of Education),Shanghai Key Laboratory of Traditional Chinese Medicine,Shanghai201203,China;Institute of Interdisciplinary Integrative Medicine Research,Shanghai University of Traditional Chinese Medicine,Shanghai201203,China)

机构地区：[1]上海中医药大学附属曙光医院,上海市中医药研究院肝病研究所,上海201203 [2]肝肾疾病病证教育部重点实验室,上海市中医临床重点实验室,上海201203 [3]上海中医药大学交叉科学研究院,上海市中医药化学生物学前沿基地,上海201203

出　　处：《中华肝脏病杂志》2024年第11期1019-1027,共9页Chinese Journal of Hepatology

基　　金：国家自然科学基金重点项目(82130120)。

摘　　要：目的探讨小鼠骨髓间充质干细胞(BMMSC)移植对不同剂量X射线致小鼠肝损伤的干预作用。方法将18只雌性C57BL/6J小鼠随机分为0、2、3 Gy照射组,0、2、3 Gy移植组:照射组作为对照,予等体积培养基注射;移植组小鼠予以不同剂量X射线辐照,并予以BMMSC骨髓内注射;第21天末处死取材。每日记录小鼠体质量变化,全自动血液检测仪检测外周血淋巴细胞、红细胞、血小板、血红蛋白含量变化,苏木精-伊红染色观察小鼠肝组织形态变化,生物化学分析仪检测小鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)活性,微板法检测肝组织还原型谷胱甘肽含量,硫代巴比妥酸法检测肝组织丙二醛含量,羟胺法检测肝组织总超氧化物歧化酶(T-SOD)含量,免疫组织化学法检测肝组织F4/80蛋白表达,蛋白免疫印迹法测定肝组织核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)蛋白表达,实时荧光定量聚合酶链式反应检测肝组织核苷酸结合结构域富含亮氨酸重复序列和含热蛋白结构域受体3(NLRP3)、白细胞介素(IL)-6、Nrf2 mRNA表达。多组比较采用析因设计方差分析,组间比较采用LSD法进行统计学分析。结果与0 Gy照射组比较,3 Gy照射组小鼠外周血淋巴细胞、红细胞、血小板、血红蛋白含量显著降低(P<0.05),血清ALT和AST活性显著升高(P<0.05);肝组织丙二醛含量、F4/80蛋白表达水平、NLRP3与IL-6 mRNA表达水平显著升高(P<0.05),肝组织T-SOD与谷胱甘肽含量、Nrf2与HO-1蛋白表达水平、Nrf2 mRNA表达水平显著下降(P<0.05)。与3 Gy照射组比较,3 Gy移植组外周血淋巴细胞、红细胞、血小板、血红蛋白含量显著升高(P<0.05),血清ALT和AST活性显著下降(P<0.05),肝组织丙二醛含量、F4/80蛋白表达水平、NLRP3与IL-6 mRNA表达水平显著降低(P<0.05),肝组织T-SOD与谷胱甘肽含量、Nrf2与HO-1蛋白表达水平、Nrf2 mRNA表达水平显著升高(P<0.05)。结论3 Gy剂量X�ObjectiveTo investigate the interventional effect of bone marrow mesenchymal stem cell(BMMSC)transplantation with different doses of X-ray irradiation induced hepatic injury in mice.MethodsEighteen female C57BL/6J mice were randomly divided into 0,2,and 3 Gy irradiation groups and 0,2,and 3 Gy transplantation groups.The irradiation group was used as the control and injected with an equal volume of culture medium.The mice in the transplantation group were irradiated with different doses of X-ray irradiation,and BMMSCs were intravenously infused into the bone marrow.The mice were sacrificed for sampling at the end of the 21st day.Mice body weight changes were recorded daily.The changes in the content of peripheral blood lymphocytes,red blood cells,platelets,and hemoglobin were detected by an automatic blood tester.The morphological changes in mice liver tissues were observed by hematoxylin-eosin staining.The serum activities of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were detected by a biochemical analyzer.The reduced glutathione contents in liver tissue were detected by the microplate method.The malondialdehyde content in liver tissue was detected by thiobarbituric acid.The content of total superoxide dismutase(T-SOD)in liver tissue was detected by the hydroxylamine method.The expression of the F4/80 protein in liver tissue was detected by the immunohistochemistry method.The protein expression of nuclear transcription factor erythroid 2 related factor 2(Nrf2)and heme oxygenase 1(HO-1)in liver tissue was determined by the western blotting method.The mRNA expression of NLRP3,IL-6,and Nrf2 in liver tissue was detected by a real-time quantitative polymerase chain reaction.The multiple-group comparisons were analyzed by factorial analysis of variance.The inter-group comparisons were analyzed by the LSD method for statistical analysis.ResultsThe contents of peripheral blood lymphocytes,erythrocytes,platelets,and hemoglobin were significantly decreased in the 3 Gy irradiation group than the 0 Gy i

关 键 词：骨髓间充质干细胞 移植 X射线 肝损伤 氧化应激 Nrf2/HO-1通路 治疗 

分 类 号：R575[医药卫生—消化系统]