腹腔注射富氢生理盐水促进高脂血症金黄地鼠胆固醇逆向转运和血脂水平降低  

Intraperitoneal injection of hydrogen-rich saline promotes reverse cholesterol transport and decreases plasma lipid levels in golden hamster with hyperlipidemia

作　　者：陈微 王新程 陈远 秦树存 CHEN Wei;WANG Xincheng;CHEN Yuan;QIN Shucun(The Second Affiliated Hospital of Shandong First Medical University,Tai’an,Shandong 271000,China;School of Clinical and Basic Medicine,Shandong First Medical University,Tai’an,Shandong 271000,China;Taishan Institute of Hydrogen Biomedical Research,Shandong First Medical University,Tai’an,Shandong 271000,China)

机构地区：[1]山东第一医科大学第二附属医院,山东省泰安市271000 [2]山东第一医科大学临床与基础医学院,山东省泰安市271000 [3]山东第一医科大学泰山氢生物医学研究院,山东省泰安市271000

出　　处：《中国动脉硬化杂志》2024年第12期1041-1050,共10页Chinese Journal of Arteriosclerosis

基　　金：国家自然科学基金项目(NSFC 81929002);山东第一医科大学学术提升计划项目(2019QL010);山东省高校重大疾病与氢医学转化应用重点实验室(鲁教科字[2022]7号);山东省医药卫生氢分子生物医学研究重点实验室(鲁卫科教字[2022]3号)。

摘　　要：[目的]观察腹腔注射富氢生理盐水(HRS)对高脂饲料饲喂金黄地鼠胆固醇逆向转运的影响。[方法]将雄性金黄地鼠分为三组:正常饮食组、高脂饮食组和高脂饮食+HRS组。高脂饲料饲喂和HRS同时持续干预12周后,将3H-胆固醇标记的巨噬细胞注射到金黄地鼠体内,检测血液、肝脏、胆汁和粪便中的放射性;通过RT-qPCR和Western blot检测肝脏组织中胆固醇逆向转运相关基因转录水平和蛋白表达水平。[结果]长期的高脂饲料饲喂诱导金黄地鼠出现明显的高脂血症和肝脏脂质蓄积。与高脂饮食组相比,高脂饮食+HRS组金黄地鼠体质量下降(P<0.01),血浆总胆固醇(TC)、低密度脂蛋白胆固醇(LDLC)显著降低(P<0.05),甘油三酯(TG)略有降低(P=0.11),高密度脂蛋白胆固醇(HDLC)显著升高(P<0.01),血浆和肝脏中氧化应激指标丙二醛(MDA)显著降低(P<0.05或P<0.01),抗氧化指标谷胱甘肽(GSH)显著升高(P<0.01),肝体比略下降(P=0.05),肝脏中TC和TG分别下降10.8%(P=0.05)和20.1%(P<0.01),肝脏脂肪变性明显缓解,但是炎症因子水平没有明显变化。同位素示踪结果显示,与高脂饮食组相比,HRS组金黄地鼠血浆3H放射性在24 h和48 h分别下降16.5%(P<0.01)和8.9%(P<0.05),肝脏、胆汁和粪便中的3H放射性分别升高了1.2倍(P<0.05)、1.2倍(P=0.08)和1.1倍(P=0.08)。RT-qPCR和Western blot结果显示,HRS干预使高脂饲料饲喂金黄地鼠肝脏CD36、清道夫受体B1(SR-B1)和低密度脂蛋白受体(LDLR)蛋白水平分别降低39.5%(P<0.05)、40.5%(P<0.01)和28.0%(P<0.01),ATP结合盒转运体G5(ABCG5)和固醇调节元件结合蛋白2(SREBP2)蛋白水平分别升高1.5倍(P<0.05)和1.3倍(P<0.01),同时ATP结合盒转运体A1和G8的mRNA水平分别升高2.9倍(P<0.05)和3.2倍(P<0.01)。[结论]腹腔注射HRS可促进高脂饲料饲喂金黄地鼠胆固醇逆向转运,并发挥降血脂效应;此外,腹腔注射HRS也可能通过抑制肝脏对脂质的摄取和促进肝脏胆固醇外排来Aim To observe the effect of intraperitoneal injection of hydrogen-rich saline(HRS)on reverse cholesterol transport in golden hamsters with high fat diet.Methods The male golden hamsters were divided into three groups:chow diet group,high-fat diet group,and high-fat diet+HRS group.After 12 weeks of high-fat diet feeding and HRS administration,macrophages labeled with 3H-cholesterol were injected into hypercholesterolemic golden hamsters,and then radioactivity in blood,liver,bile,and feces was measured.RT-qPCR and Western blot were used to assess the transcription and protein expression levels of cholesterol reverse transport-related genes in liver tissue.Results Long-term feeding with a high-fat diet induced significant hyperlipidemia and liver lipid accumulation in golden hamsters.Compared with the high-fat diet group,after HRS intervention,the body mass of golden hamsters decreased(P<0.01),plasma TC and LDLC significantly decreased(P<0.05),TG slightly decreased(P=0.11),HDLC significantly increased(P<0.01),oxidative stress index MDA in plasma and liver significantly decreased(P<0.05 or P<0.01),antioxidant index glutathione(GSH)significantly increased(P<0.01),liver weight/body weight ratio slightly decreased(P=0.05),TC and TG in liver decreased by 10.8%(P=0.05)and 20.1%(P<0.01),respectively.Liver steatosis was significantly relieved,but there was no significant change in inflammatory factor levels.In isotopic tracing,high-fat diet fed golden hamsters treated with HRS showed decreased 3H radioactivity in plasma at 24 and 48 hours by 16.5%(P<0.01)and 8.9%(P<0.05)respectively,while increased 3H radioactivity was observed in liver,bile,and feces by 1.2-fold(P<0.05),1.2-fold(P=0.08),and 1.1-fold(P=0.08)respectively,compared to those fed a high-fat diet alone.Furthermore,RT-qPCR and Western blot analyses of liver tissue demonstrated that HRS intervention resulted in a decrease of CD36,scavenger receptor-B1(SR-B1),and low density lipoprotein receptor(LDLR)protein levels by 39.5%(P<0.05),40.5%(P<0.01),and 28.0%(P<0.01)re

关 键 词：富氢生理盐水 高脂血症 金黄地鼠 胆固醇逆向转运 血脂 

分 类 号：R5[医药卫生—内科学] R363[医药卫生—临床医学]