摘要
目的:研究电针"肝俞""阳陵泉"对大鼠肝脏缺血再灌注损伤(HIRI)的保护作用和对肝脏组织高迁移率族蛋白1(HMGB1)表达的影响。方法:雄性SD大鼠随机分为假手术组、模型组、肝俞-阳陵泉组和非经非穴组,每组10只。采用无创动脉夹阻断供应肝中叶和左叶的动脉、静脉和胆管1 h,解除阻断后再灌注4 h以构建70%HIRI模型。肝俞-阳陵泉组取两侧"肝俞"和"阳陵泉",非经非穴组取两侧"肝俞"穴区旁开6~8 mm的非经非穴区肌肉结缔组织,此二组均在造模前给予电针刺激30 min。以HE染色法观察肝脏组织损伤情况,全自动生化分析仪检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)水平,酶联免疫法检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和HMGB1水平,免疫组织化学法和蛋白质免疫印迹法检测肝脏组织HMGB1蛋白表达,荧光定量PCR检测肝脏组织HMGB1 mRNA表达。结果:与假手术组比较,造模后大鼠肝脏损伤评分及血清ALT、AST、TNF-α、IL-6和HMGB1含量均显著升高(P<0.01),肝脏组织中的HMGB1阳性细胞数、HMGB1 mRNA及蛋白表达均明显升高(P<0.01)。与模型组比较,肝俞-阳陵泉组大鼠肝脏损伤评分及血清ALT、AST、TNF-α、IL-6和HMGB1含量均降低(P<0.05),肝脏组织中的HMGB1阳性细胞数、HMGB1 mRNA及蛋白表达降低(P<0.05)。非经非穴组大鼠肝脏损伤评分及血清ALT、AST、TNF-α、IL-6和HMGB1含量均高于肝俞-阳陵泉组(P<0.05),肝脏组织中HMGB1阳性细胞数、HMGB1 mRNA及蛋白表达均高于肝俞-阳陵泉组(P<0.05)。结论:电针"肝俞""阳陵泉"可减轻大鼠肝脏缺血再灌注损伤,这种保护作用可能是通过抑制HMGB1从胞核向胞质移位和释放、下调相关炎性因子表达来发挥的。
Objective To explore the protective effect of electroacupuncture(EA)on hepatic ischemia-reperfusion injury(HIRI)and the expression of high mobility group protein 1(HMGB1)in liver tissues in rats.Methods A total of 40 male SD rats were randomly divided into 4 groups,namely sham control,HIRI model,"Ganshu"(BL18)-"Yanglingquan"(GB34)and non-acupoint group,with 10 rats in each group.The HIRI model was induced by blocking the arteries,veins and bile ducts supplying the middle and left lobes of the liver for 1 h,and reperfusion for 4 h to induce an area of about 70%HIRI.EA was applied to bila-teral BL18 and GB34,or non-acupoints about 6—8 mm to the bilateral BL18 for 30 min before modeling.Serum alanine transaminase(ALT)and aspartate aminotransferase(AST)levels were measured by using an automatic biochemical analyzer.Serum tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and HMGB1 levels were assayed by ELISA.Hematoxylin-eosin(H.E.)staining was used to observe histopathological changes of the liver tissue by using tissue injury scaling(0-3 scores).The expression of HMGB1 protein in liver tissues was detected by immunohistochemical staining,Western blot and PCR,separately.Results Following modeling and compared with the sham group,the levels of serum ALT,AST,TNF-α,IL-6,and HMGB1 contents,the number of HMGB1 immunoreaction(IR)-positive cells,and HMGB1 protein and mRNA were significantly increased(P<0.01).After the treatment,the contents of serum ALT,AST,TNF-α,IL-6,and HMGB1,liver HMGB1 IR-positive cells,protein and mRNA were considerably down-regulated in the BL18-GB34 group(P<0.05),rather than in the non-acupoint group(P>0.05)in contrast to the model group.H.E.stain showed a higher liver injury score in the model group than in the sham group(P<0.01),and a lower liver injury score in the BL18-GB34 group(not the non-acupoint group)relevant to the model group(P<0.05).Conclusion EA of BL18 and GB34 points has a protective effect on ischemic liver injury in rats with HIRI,which may be associated with its functions in inhibiting the migration and release of HMGB1 from the nucleus to the cytoplasm and in down-regulating the expression of inflammatory factors.
作者
谭思由
黄俊
苏颖颖
陈文雁
孔高茵
刘景诗
魏来
TAN Si-you;HUANG Jun;SU Ying-ying;CHEN Wen-yan;KONG Gao-yin;LIU Jing-shi;WEI Lai(Department of Anesthesiology,Hunan Provincial People’s Hospital/The First Affiliated Hospital of Hunan Normal University,Changsha 410002,China;Hunan Province Perioperative Accelerated Rehabilitation Anesthesia Clinical Medical Research Center,Changsha 410002)
出处
《针刺研究》
CAS
CSCD
北大核心
2020年第11期888-894,共7页
Acupuncture Research
基金
湖南省教育厅项目(No.16C0971)
湖南省中医药管理局项目(No.202071)。
关键词
电针
肝脏缺血再灌注损伤
高迁移率族蛋白1
炎性因子
Electroacupuncture
Hepatic ischemia-reperfusion injury
High mobility group protein 1
Inflammatory factor
