摘要
目的观察吴茱萸次碱对小鼠肝脏缺血再灌注损伤的保护作用并探讨其作用机制。方法将24只成年雄性C57BL/6小鼠按随机数表法随机分为假手术组(Sham组)、70%肝脏缺血再灌注损伤组(HIRI组)及治疗组。采用夹闭肝左叶、肝中叶肝蒂方法建立肝脏缺血再灌注损伤模型,Sham组于手术前连续3d腹腔注射等量生理盐水然后再做开腹处理,HIRI组给予等量生理盐水再建模,治疗组则给予504μg/(kg·d)吴茱萸次碱后再建模。夹闭60 min再灌注6 h后处死小鼠,收集外周血及肝组织。全自动血清生化分析仪检测外周血丙氨酸氨基转移酶(ALT)及天冬氨酸氨基转移酶(AST)水平;酶联免疫吸附试验(ELISA)检测肝组织中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6及IL-1β含量;化学比色法检测肝组织中丙二醛(MDA)水平及超氧化物歧化酶(SOD)活性;苏木精-伊红(HE)染色检测肝组织病理变化;定量聚合酶链反应(qPCR)法检测肝组织中Toll样因子受体4(TLR4)及核因子-κB(NF-κB)的mRNA表达;蛋白质印迹法(Western blot)检测肝组织中TLR4及NF-κB的蛋白表达。采用单因素方差分(One-Way ANOVA)和LSD-t检验对数据进行检验。结果再灌注6 h后,治疗组小鼠外周血ALT及AST低于HIRI组[ALT:(166.200±9.682)U/L比(303.400±9.882)U/L,F=203.000,P<0.01;AST:(313.200±14.360)U/L比(504.400±11.740)U/L,F=304.000,P<0.01];治疗组肝组织中炎症因子TNF-α、IL-6及IL-1β低于HIRI组[TNF-α:(196.800±15.540)pg/mg比(304.200±24.960)pg/mg,F=32.950,P<0.01;IL-6:(480.800±32.910)pg/mg比(760.400±61.270)pg/mg,F=44.770,P<0.01;IL-1β:(623.600±73.300)pg/mg比(958.400±117.300)pg/mg,F=15.620,P<0.01];治疗组肝组织中MDA低于HIRI组[(4.996±0.161)nmol/mg比(6.804±0.256)nmol/mg,F=58.810,P<0.01];治疗组肝组织中SOD活性高于HIRI组[(71.800±1.590)U/mg比(50.100±1.292)U/mg,F=71.070,P<0.01]。治疗组肝组织中肝窦与中央静脉充血、肝细胞肿胀变性坏死及炎细胞聚集等病理改变轻于
Objective To investigate the protective effect of Rutaecarpine(Rut)against hepatic ischemia-reperfusion injury(HIRI)in mice and its mechanism.Methods Totally,24 adult male C57BL/6 mice were randomly divided into three groups:Sham group,HIRI group and treatment group(n=8 each).The animals in treatment group were injected intraperitoneally with Rut[504μg/(kg·d)]before operation.Except for Sham group,an 70%volume HIRI model was established by 60 min ischemia and then 6 h reperfusion in the other two groups.The peripheral blood and liver tissues were collected.The serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were determined by automatic biochemistry analyzer.The protein expression levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-1βin hepatic tissues were measured by enzyme linked immunosorbent assay(ELISA).The levels of malondialdehyde(MDA)and enzyme activities of superoxide dismutase(SOD)were measured by chemical colorimetry.The pathological changes in liver tissues were examined by hematoxylin and eosin(HE)staining.The mRNA and protein expression of Toll-like receptor 4(TLR4)and nuclear factor-κB(NF-κB)was detected by quantitative polymerase chain reaction(qPCR)and Western blotting,respectively.Data were analyzed by one-way ANOVA and LSD-t.Results At 6 h after reperfusion,the serum levels of ALT and AST in treatment group were both remarkably lower than those in HIRI group[ALT:(166.200±9.682)vs.(303.400±9.882)U/L,F=203.000,P<0.01;AST:(313.200±14.360)vs.(504.400±11.740)U/L,F=304.000,P<0.01].The levels of TNF-α,IL-6 and IL-1βin treatment group were both lower than those in HIRI group[TNF-α:(196.800±15.540)vs.(304.200±24.960)pg/mg,F=32.950,P<0.01;IL-6:(480.800±32.910)vs.(760.400±61.270)pg/mg,F=44.770,P<0.01;IL-1β:(623.600±73.300)vs.(958.400±117.300)pg/mg,F=15.620,P<0.01].The expression of MDA in treatment group was lower than that in HIRI group[(4.996±0.161)vs.(6.804±0.256)nmol/mg,F=58.810,P<0.01].The enzyme activity of SOD in treatment group w
作者
邹雯佳
游建
汪洁
王夏
何鑫
Zou Wenjia;You Jian;Wang Jie;Wang Xia;He Xin(Department of General Surgery,Wuhan Forth Hospital Puai Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430033,China)
出处
《中华实验外科杂志》
CAS
北大核心
2022年第5期865-868,共4页
Chinese Journal of Experimental Surgery
