苓桂术甘汤调控自噬对多柔比星抗非酒精性脂肪肝病相关肝细胞癌的增效作用  

Synergistic effects of Linggui zhugan decoction regulating autophagy on doxorubicin against non-alcoholic fatty liver disease-hepatocellular carcinoma

作　　者：曹慧敏[1] 孔亮[1,2] 隋国媛 吴瑾[1] 贾连群[1] CAO Huimin;KONG Liang;SUI Guoyuan;WU Jin;JIA Lianqun(Key Laboratory of Theory and Application of Visceral State of Traditional Chinese Medicine Co-constructed by Ministry of Education,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;School of Pharmacy,Liaoning University of Traditional Chinese Medicine,Liaoning Dalian 116600,China)

机构地区：[1]辽宁中医药大学中医脏象理论及应用教育部重点实验室,沈阳110847 [2]辽宁中医药大学药学院,辽宁大连116600

出　　处：《中国药房》2023年第19期2316-2322,共7页China Pharmacy

基　　金：国家自然科学基金项目(No.82204629);辽宁省教育厅高校基本科研项目(No.LJKQZ20222384);辽宁省自然科学基金项目(No.2022-BS-197)。

摘　　要：目的探讨苓桂术甘汤(LGZG)通过调控自噬增强多柔比星(DOX)的抗非酒精性脂肪肝病相关肝细胞癌(NAFLD-HCC)的作用及机制。方法将C57BL/6小鼠采用随机数字表法分为空白组、NAFLD-HCC组、LGZG组、DOX组、DOX+LGZG组,每组10只。采用腹腔注射二乙基亚硝胺溶液(50mg/kg)和喂养高脂饲料相结合的方法建立NAFLD-HCC小鼠模型;空白组小鼠注射等量生理盐水,给予普通饲料喂养。建模后,各给药组分别灌胃LGZG水提液(20g/kg)和/或腹腔注射DOX溶液(8mg/kg),空白组、NAFLD-HCC组灌胃等量生理盐水,每日1次,连续4周。在模型建立及药物干预期间,监测小鼠的一般情况。药物干预结束后,检测小鼠体重、肝重及肝脏系数;观察小鼠肝脏组织病理形态和纤维化程度;检测小鼠血脂水平[甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)的水平],血清中甲胎蛋白(AFP)、癌胚抗原(CEA)含量,肝脏组织中增殖标志物Ki-67、B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)的表达水平,以及微管相关蛋白1A/1B-轻链3(LC3)、苄氯素1(Beclin1)、选择性自噬接头蛋白P62的蛋白表达水平。结果与空白组比较,NAFLD-HCC组小鼠随着给药时间延长活动逐渐减少,精神萎靡,毛发蓬乱无光泽,体重显著降低(P<0.05),肝重有上升趋势,肝脏系数显著增加(P<0.05);小鼠肝脏组织炎症细胞大量浸润,部分细胞出现气球样变及小细胞性增生的病变,肝脏纤维化程度加重;血清中TC、TG、LDL-C水平及AFP、CEA含量均显著升高,HDL-C水平显著降低(P<0.05);肝脏组织中Ki-67、Bcl-2蛋白表达水平均显著升高(P<0.05),Bax蛋白表达水平有降低趋势;肝脏组织中Beclin1蛋白表达水平显著降低(P<0.05),LC3Ⅱ/LC3Ⅰ比值有所降低,P62蛋白表达水平有所升高。与NAFLD-HCC组比较,DOX组、LGZG组及DOX+LGZG组小鼠上述指标均有不同程度改善,且DOX联合LGZG的干预效果优于DOX。结�OBJECTIVE To explore the enhancement effect of Linggui zhugan decoction(LGZG)regulating autophagy on doxorubicin(DOX)against non-alcoholic fatty liver disease-hepatocellular carcinoma(NAFLD-HCC).METHODS C57BL/6 mice were randomly divided into blank group,NAFLD-HCC group,LGZG group,DOX group and DOX+LGZG group,with 10 mice in each group.The NAFLD-HCC model was established by intraperitoneal injection of diethylnitrosamine(50 mg/kg)and high-fat diet.The blank group was injected with the same amount of normal saline and fed with ordinary diet.After modeling,administration groups were given LGZG aqueous extract(20 g/kg)intragastrically and/or DOX solution intraperitoneally(8 mg/kg);the blank group and NAFLD-HCC group were given a constant volume of normal saline intragastrically,once a day,for 4 consecutive weeks.The general condition of mice was monitored during modeling and drug intervention.After drug intervention,body weight,liver weight and liver coefficient of mice were detected.The histopathologic morphology and fibrosis degree of liver tissue in mice were observed;the levels of blood lipid[the levels of triglyceride(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)],the serum contents of alpha fetoprotein(AFP)and carcinoembryonic antigen(CEA),and the expressions of marker of proliferation Ki-67,B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X(Bax)in liver tissue were all detected as well as protein expressions of microtubule-associated proteins 1A and 1B(LC3),Beclin1 and selective autophagy adopt proteins P62.RESULTS Compared with the blank group,the activity of mice decreased gradually as time in the NAFLD-HCC group;mental fatigue,disheveled and matte hair were observed,and body weight decreased significantly(P<0.05);liver weight had an upward trend,and liver coefficient increased significantly(P<0.05).The inflammatory cells of liver tissue were infiltrated,with some cells showing ballooning and small cell hyperplasia,and the degree of liver fib

关 键 词：苓桂术甘汤 非酒精性脂肪肝病相关肝细胞癌 自噬 多柔比星 化疗增敏 

分 类 号：R965[医药卫生—药理学]