摘要
目的探索奥贝胆酸(OCA)和白藜芦醇(RSV)联合应用对非酒精性脂肪性肝病(NAFLD)模型小鼠的治疗作用和机制。方法雄性C57BL/6N小鼠42只,除正常对照组8只外,其余用1%四氯化碳(CCl_(4))按5 mL·kg^(-1)每周ip注射1次联合高脂饲料喂食4周诱导NAFLD模型。建模小鼠分为模型对照组、模型+OCA组、模型+RSV组和模型+OCA+RSV组,OCA和RSV均30 mg·kg^(-1) ig给药连续28 d。小鼠处死取血制备血清,取肝称重计算肝指数,HE染色观察肝组织病理形态,油红O染色检测肝细胞脂质沉积并统计脂滴面积百分比;全自动生化分析仪和检测试剂盒检测血清谷丙转氨酶(GPT)、谷草转氨酶(GOT)、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)、超氧化物歧化酶(SOD)活性、丙二醛(MDA)、白细胞介素1β(IL-1β)、IL-6和肿瘤坏死因子α(TNF-α)水平;Western印迹法检测肝组织沉默信息调节因子1(Sirt1)、NF-κB和p-NF-κB蛋白表达水平。结果与正常对照组相比,模型组小鼠肝指数显著升高(P<0.01);与模型组相比,各给药组肝指数显著降低(P<0.01);与模型+RSV相比,模型+OCA+RSV组肝指数降低更加明显(P<0.05)。与正常对照组相比,模型组小鼠肝组织有大量脂肪空泡和炎症损伤,油红O染色后细胞内有大量红色脂滴,血清GPT,GOT,TC,LDL,MDA,IL-1β,IL-6和TNF-α水平及肝组织NF-κB蛋白表达和磷酸化水平显著升高(P<0.01),血清HDL水平和肝组织Sirt1表达水平显著降低(P<0.05)。与模型组相比,模型+OCA组肝细胞形态正常,肝组织少量脂肪变性空泡,红色脂滴减少,血清GOT,TC,MDA,IL-6和TNF-α水平显著降低(P<0.05),GTP,HDL和LDL水平无明显变化;模型+RSV组脂滴空泡无明显减少,细胞内红色脂滴略有减少,血清TC,LDL,MDA,IL-1β,IL-6和TNF-α水平显著降低(P<0.05),GPT,GOT和HDL水平无明显变化,肝组织p-NF-κB蛋白表达水平显著降低(P<0.05),肝组织Sirt1表达水平显著升高(P<0.05);模型+OCA+RSV
OBJECTIVE To explore the therapeutic effect and mechanism of obeticholic acid(OCA)combined with resveratrol(RSV)in model mice with non-alcoholic fatty liver disease(NAFLD).METHODS Forty-two male C57BL/6N mice,except the normal control group,were injected with 1% CCl_(4) by ip 5 mL·kg^(-1) once a week and fed with high-fat diet for four weeks to induce an NAFLD model.The model mice were divided into the model group,model+OCA group,model+RSV group and model+OCA+RSV group,and ig administered with or without OCA 30 mg·kg^(-1) or(and)RSV 30 mg·kg^(-1),once a day,for 28 d.The mice were sacrificed,blood was collected,and the serum was collected by centrifugation.The liver tissues were collected to calculate the liver index.The pathological changes of the liver tissue of the mice were observed by HE staining.The serum levels of glutamic pyruvic transaminase(GPT),glutamic oxaloacetic transaminase(GOT),total cholesterol(TC),triglycerides(TG),low density lipoprotein(LDL),high density lipoprotein(HDL),superoxide dismutase(SOD),malondialdehyde(MDA),interleukin-1β(IL-1β),IL-6 and tumor necrosis factor-α(TNF-α)were detected by automatic biochemical analyzer and detection kits.The protein expression levels of sirtuin 1(Sirt1),NF-κB and p-NF-κB in liver tissue were detected by Western blotting.RESULTS Compared with the normal control group,the liver indexes of model group were increased significantly(P<0.01).Compared with the model group,the liver indexes of drug treated groups were decreased significantly(P<0.01).Compared with the model+RSV group,the liver indexes of the model+OCA+RSV group were decreased significantly(P<0.05).The liver tissue sections of the mice in the model group had a large number of fat vacuoles and inflammatory lesions,and a large number of red fat deposits were found via oil red O staining.The levels of serum GPT,GOT,TC,LDL,MDA,IL-1βand IL-6,the level of TNF-αand the expression and phosphorylation of NF-κB in liver tissue were significantly increased(P<0.01),while the level of serum HDL and th
作者
高川
姚宜山
周奎臣
付海洋
王金柱
梁霞霞
王瑜
GAO Chuan;YAO Yi-shan;ZHOU Kui-chen;FU Hai-yang;WANG Jin-zhu;LIANG Xia-xia;WANG Yu(Department of Clinical Medicine,Jiamusi University,Jiamusi 154007,China;Institute of Pharma-cology and Toxicology,Academy of Military Medial Sciences,Beijing 100850,China;Department of Pharmacy,Changzhi Medical College,Changzhi 046000,China)
出处
《中国药理学与毒理学杂志》
CAS
北大核心
2022年第5期338-345,共8页
Chinese Journal of Pharmacology and Toxicology
基金
国家自然科学基金(21302223)
佳木斯大学校长创新创业基金(XZFY2018-43)。
关键词
奥贝胆酸
非酒精性脂肪肝
白藜芦醇
氧化应激
obeticholic acid
non-alcoholic fatty liver
resveratrol
oxidative stress
