当飞利肝宁胶囊对CCl4诱导的非酒精性脂肪性肝病大鼠急性肝损伤肝组织SIRT1/NF-κB/p53信号通路的影响  

The effect of Dangfei Liganning Capsules on the SIRT1/NF-κB/p53 signaling pathway in liver tissues of CCl4-induced acute liver injury in rats with non-alcoholic fatty liver disease

作　　者：周岐鸣 宋海燕 刘洋 续嗣钰 杨丽丽 柳涛 王倩蕾 ZHOU Qiming;SONG Haiyan;LIU Yang;XU Siyu;YANG Lii;LIU Tao;WANG Qianei(Institute of Digestive Diseases,Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Department of Nephrology,Lanxi Hospital of Traditional Chinese Medicine,Lanxi,Zhejiang 321100,China;Second Department of Digestive Diseases,Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201316,China)

机构地区：[1]上海中医药大学附属龙华医院脾胃病研究所,上海200032 [2]兰溪市中医院肾内科,浙江兰溪321100 [3]上海中医药大学附属龙华医院脾胃病二科,上海201316

出　　处：《上海中医药杂志》2024年第5期78-85,共8页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82274386)。

摘　　要：目的基于沉默信息调节因子1(SIRT1)调控的炎症和凋亡相关信号通路,探讨当飞利肝宁胶囊改善四氯化碳(CCl4)诱导的非酒精性脂肪性肝病(NAFLD)大鼠急性肝损伤的作用机制。方法雄性Wistar大鼠按体质量随机分为正常(N)组、NAFLD模型(HF)组、正常加CCl4(N-CCl4)组、NAFLD加CCl4(HF-CCl4)组和当飞利肝宁胶囊(DF)组。N和N-CCl4组大鼠给予普通饲料,HF、HF-CCl4组及DF组均给予高脂饲料,DF组同时予当飞利肝宁胶囊(0.3 g·kg^(-1))每天灌胃干预。8周后,除N和HF组外,其余组大鼠腹腔注射小剂量CCl4。48 h后取大鼠血清和肝组织。检测指标包括:①通过苏木精-伊红(HE)染色观察肝组织病理,检测血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST);②通过实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测肝组织促炎因子白介素-1β(IL^(-1β))、单核细胞趋化蛋白1(MCP1)及肿瘤坏死因子-α(TNF-α)的mRNA水平,同时采用Western blot法检测肝组织组蛋白-赖氨酸N-甲基转移酶(SUV39H2)、SIRT1、乙酰化核因子-κB亚基p65(Acetyl-p65)、乙酰化肿瘤抑制蛋白p53(Acetyl-p53)、剪切型多聚ADP核糖多聚酶(Cleaved-PARP)及凋亡相关分子p53上调凋亡调节因子(PUMA)、Bcl-2相关蛋白(Bcl-xL)的蛋白表达水平。结果①HF组大鼠肝组织出现广泛肝细胞脂肪变性,HF-CCl4组除此之外还出现大量肝细胞气球样变性、炎症细胞浸润和凋亡小体,DF组大鼠肝组织脂肪变性、气球样变和炎症浸润程度均显著减轻。小剂量CCl4未导致肝组织异常改变。血清ALT、AST水平在各组大鼠中的变化趋势同肝组织病理改变,在HF组比N、N-CCl4组上调(P<0.05),在HF-CCl4组又较HF组增加(P<0.05),而在DF组上述转氨酶下调(P<0.05)。②肝组织SIRT1水平在HF组和HF-CCl4组大鼠低于N组(P<0.05),DF组SIRT1水平显著提高(P<0.05);SUV39H2表达水平变化则与SIRT1趋势相反(P<0.05)。大鼠肝组织Acetyl-p65水平及促炎因子IL^(-1β)、MObjective To investigate the mechanism of Dangfei Liganning Capsules in improving acute liver injury induced by carbon tetrachloride(CCl4)in rats with non-alcoholic fatty liver disease(NAFLD)based on the inflammation and apoptosis-related signaling pathways regulated by Sirtuin 1(SIRT1).Methods Male Wistar rats were randomly divided into a normal(N)group,an NAFLD model(HF)group,a normal with CCl4(N-CCl4)group,an NAFLD with CCl4(HF-CCl4)group,and a Dangfei Liganning Capsule(DF)group.Rats in the N and N-CCl4 groups were fed a normal standard diet,while those in the HF,HF-CCL4,and DF groups received a high-fat diet.Additionally,the DF group was simultaneously treated with Dangfei Liganning Capsules(0.3 g·kg^(-1))daily by gavage.After 8 weeks,except for the N and HF groups,the remaining rats received an intraperitoneal injection of a low dose of CCl4.Rat serum and liver tissues were collected 48 hours later.The parameters measured included:①Liver tissue pathology was observed through HE staining;Serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels were tested;②mRNA levels of pro-inflammatory cytokines interleukin-1β(IL^(-1β)),monocyte chemoattractant protein 1(MCP1)and tumor necrosis factor-α(TNF-α)in liver tissues were detected by RT-qPCR,while protein expression levels of suppressor of variegation 3-9 homolog 2(SUV39H2),SIRT1,acetylated p65(Acetyl-p65),acetylated p53(Acetyl-p53),cleaved poly ADP-ribose polymerase(Cleaved-PARP),P53 up-regulated modulator of apoptosis(PUMA)and Bcl-2 associated protein(Bcl-xL)were measured by Western blot.Results①Diffuse hepatic steatosis was observed in the HF group,while the HF-CCl4 group also showed significant hepatocyte ballooning,inflammatory cell infiltration,and apoptotic bodies.The degree of steatosis,ballooning,and inflammatory infiltration in the DF group was significantly alleviated.Low-dose CCl4 did not cause abnormal pathological changes in liver tissues.The change trends in serum ALT and AST levels among the rats in each group correspo

关 键 词：非酒精性脂肪性肝病 肝损伤 当飞利肝宁胶囊 大鼠模型 作用机制 中药研究 

分 类 号：R285.5[医药卫生—中药学]