摘要
目的探究烟酰胺腺嘌呤二核苷酸(NADH)通过SIRT1/Nrf2通路调控小鼠抗结核药物性肝损伤和凋亡的机制。方法将24只6周龄SPF级雄性小鼠按体质量随机分成4组:ADLI组[90 mg/(kg·d)异烟肼+135 mg/(kg·d)利福平+315 mg/(kg·d)吡嗪酰胺灌胃]、对照组[与抗结核药物性肝损伤(ADLI)组等体积生理盐水灌胃]、NADH组(对照组基础上应用30 mg/kg NADH灌胃)和NADH干预组(ADLI组基础上应用30 mg/kg NADH灌胃),每组6只,连续灌胃7 d,收集血清和肝组织。qRT-PCR法和Western blot法分别检测SIRT1/Nrf2通路中的沉默信息调节因子1(SIRT1)、核因子E2相关因子2(Nrf2)和凋亡相关指标B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、人胱天蛋白酶3(caspase-3)的mRNA和蛋白表达情况;HE染色法观察小鼠肝脏组织形态;称取小鼠肝脏质量,将其质量除以体质量获得肝脏指数;微板法检测肝损伤指标谷氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)、乳酸脱氢酶(LDH)的水平。结果与对照组相比,ADLI组SIRT1和Nrf2蛋白、mRNA表达水平下降(P<0.05);肝组织结构紊乱,细胞明显肿胀、边界不清;小鼠体质量下降,肝指数上升;抗凋亡因子Bcl-2 mRNA和蛋白表达水平降低,凋亡因子Bax、caspase-3 mRNA和蛋白表达增强;肝损伤指标ALT、AST、LDH水平升高(P<0.05)。而与ADLI组相比,NADH干预后,SIRT1、Nrf2的mRNA和蛋白表达升高;肝组织结构清晰,细胞呈多边形;抗凋亡因子Bcl-2 mRNA和蛋白表达升高,凋亡因子Bax、caspase-3 mRNA和蛋白水平降低;小鼠体质量上升,肝指数下降;肝损伤指标ALT、AST、LDH表达降低(P<0.05)。结论NADH可能通过调控SIRT1/Nrf2通路缓解小鼠抗结核药物性肝损伤和细胞凋亡。
Objective To investigate the mechanism by which Nicotinamide adenine dinucleotide(NADH)regulates anti-tuberculosis drug-induced liver injury and apoptosis in mice through SIRT1/Nrf2 pathway.Methods Twenty-four six-week-old SPF male mice were randomly divided into four groups according to body weight,ADLI group[90 mg/(kg·d)Isoniazid,135 mg/(kg·d)Rifampicin,315 mg/(kg·d)Pyrazinamide were given by gavage],control group[thesame volume of saline was given by gavage as antituberculosis drug-induced liver injury(ADLI)group],NADH group(30 mg/kg NADH wasgiven by gavage on the basis of control group)and NADH intervention group(30 mg/kg NADH wasgiven by gavage on the basis of ADLI group),with sixmice in each group.They were gavaged continuously for seven days,and their seruand liver tissues were collected.The mRNA and protein expression of silence information regulator 1(SIRT1),nuclear factor erythroid 2-related factor 2(Nrf2)in SIRT1/Nrf2 pathway,apoptosis indicators B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax)and caspase-3 were detected by qRT-PCR and Western blot,respectively.HE staining was performed to observe the morphology of liver tissue.The liver was weighedandthe liver index was obtained by dividingweight by body weight.The levels of glutamate aminotransferase(ALT),aspartate aminotransferase(AST)and lactate dehydrogenase(LDH),which are indicators of liver injury,were detected by microplate method.Results Compared with control group,the protein and mRNA expression of SIRT1,Nrf2decreased significantly in ADLI group.Liver tissue structure wasdisturbed,hepatocytes were obviously swollen,and their boundary was unclear.The weight of mice decreased,but liver index increased.The mRNA and protein expression level of anti-apoptotic factor Bcl-2 decreased,while that of Bax and caspase-3 was raised.The level of ALT,AST and LDH were also elevated.The differences above were statistically significant(P<0.05).Compared with ADLI group,the protein and mRNA expression of SIRT1,Nrf2 were higher after NADH intervention.
作者
李金凤
崔梦祥
龙奕妃
孟春燕
任琦
冯福民
Li Jinfeng;Cui Mengxiang;Long Yifei;Meng Chunyan;Ren Qi;Feng Fumin(School of Public Health,North China University of Science and Technology,Tangshan 063210)
出处
《安徽医科大学学报》
CAS
北大核心
2023年第12期2089-2094,共6页
Acta Universitatis Medicinalis Anhui
基金
河北省自然科学基金(编号:H2021209052)
河北省高等学校基本科研业务费项目(编号:JQN2021032)。
关键词
抗结核药物性肝损伤
凋亡
炎症
烟酰胺腺嘌呤二核苷酸
沉默信息调节因子1
核因子E2相关因子2
antituberculosis drug-induced liver injury
apoptosis
inflammation
nicotinamide adenine dinucleotide
silence information regulator 1
nuclear factor erythroid 2-related factor 2
