摘要
[目的]阐明健脾祛湿方(Jianpi Qushi decoction,JQD)治疗大鼠非酒精性脂肪肝(nonalcoholic fatty liver disease,NAFLD)的药效和分子机制。[方法]试验通过饲喂高脂饲料建立大鼠NAFLD模型,将大鼠随机分为模型、奥利司他(32.4 mg/kg)和JQD低(0.35 g/kg)、高(0.70 g/kg)剂量组,另设对照组,每组6只。奥利司他和JQD不同剂量组大鼠灌胃给药,1 mL/只,每天1次,连续7周,对照组与模型组大鼠灌胃等剂量的生理盐水。试验结束后采集大鼠血液、肝脏组织。通过ELISA法测定大鼠血清血脂水平,主要包括总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C);统计大鼠肝脏指数,检测肝脏抗氧化指标(谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA));采用HE染色和油红O观察大鼠肝脏组织形态结构。利用药物和疾病靶点相关数据库分析JQD治疗NAFLD的活性成分和关键靶点;通过Cytoscape 3.7.2软件构建“药物-成分-靶点”网络图;通过分子对接和实时荧光定量PCR验证JQD主要活性成分和关键靶点的作用效果。[结果]与对照组相比,模型组大鼠血清中TC、TG、LDL-C和肝脏中MDA含量及肝脏指数均显著或极显著升高(P<0.05;P<0.01),血清中HDL-C含量和肝脏中GSH含量及SOD、CAT活性均显著或极显著降低(P<0.05;P<0.01)。与模型组比,JQD低、高剂量组大鼠血清中TC、TG含量和肝脏中MDA含量及肝脏指数均显著降低(P<0.05),肝脏中SOD、CAT活性和GSH含量均显著升高(P<0.05)。HE染色和油红O染色结果显示,对照组大鼠肝细胞形态正常,肝索整齐;模型组大鼠肝脏发生明显的脂肪变形,肝细胞萎缩,布满大小不一的空泡,肝索形态紊乱;JQD低、高剂量组大鼠肝脏空泡数量明显减少,肝索清晰可见。网络药理学共筛选出JQD的有效活性成分35个和NAFLD疾病靶点1890个;网络拓扑分析结果显示,JQD治疗NAFLD的核心活性成分为山柰酚、异鼠李
[Objective]The purpose of this experiment was to elucidate the pharmacological efficacy and molecular mechanism of Jianpi Qushi decoction(JQD)in the treatment of non-alcoholic fatty liver disease(NAFLD)in rats.[Method]After the rat NAFLD model was established by feeding high-fat diet,the rats were randomly divided into model,orlistat(32.4 mg/kg),JQD low(0.35 g/kg)and high(0.70 g/kg)dose groups,and control group was set up,with 6 rats in each group.Rats in orlistat and JQD different dose groups were given intragastric administration,1 mL/rat,once a day for 7 weeks,and the rats in control and model groups were given intragastric administration of equal dose of normal saline.After the experiment,the blood and liver tissues of rats were collected.Serum lipid levels of rats were determined by ELISA,including total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL-C)and high density lipoprotein(HDL-C).The liver index of rats was measured and the liver antioxidant indexes,including glutathione(GSH),superoxide dismutase(SOD),catalase(CAT)and malondialdehyde(MDA)were detected.HE staining and oil red O staining were used to observe the morphological structure of liver.The active ingredients and key targets of JQD for NAFLD were analyzed using drug and disease target database.The“drug-ingredient-target”network diagram was constructed by Cytoscape 3.7.2 software.The effects of main active ingredients of JQD and key targets were verified by molecular docking and Real-time quantitative PCR.[Result]Compared with control group,the contents of TC,TG and LDL-C in serum,MDA content in liver and liver index in model group were significantly or extremely significantly increased(P<0.05 or P<0.01),HDL-C content in serum,GSH content,SOD and CAT activities in liver were significantly or extremely significantly decreased(P<0.05 or P<0.01).Compared with model group,the contents of TC and TG in serum,MDA content in liver and liver index of rats in JQD low and high dose groups were significantly decreased(P<0.05),and the activ
作者
苏圆圆
于澄元
张密霞
张艳军
庄朋伟
SU Yuanyuan;YU Chengyuan;ZHANG Mixia;ZHANG Yanjun;ZHUANG Pengwei(College of Chinese Materia Medica,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;First Teaching Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China;National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion,Tianjin 300193,China)
出处
《中国畜牧兽医》
CAS
CSCD
北大核心
2024年第8期3662-3675,共14页
China Animal Husbandry & Veterinary Medicine
基金
国家重点研发计划(2018YFC1706804)。
