大柴胡汤加减方对湿热证胆囊胆固醇结石小鼠胆固醇代谢基因SRBI、ABCG5/G8表达的影响  

Influence of Modified Dachaihu Decoction on the Expression of Cholesterol Metabolism Genes SRBI,ABCG5/G8 in Cholesterol Gallstone Mice with Damp-Heat Syndrome

作　　者：王素英 章小燕 闵莉[1,2,3] WANG Suying;ZHANG Xiaoyan;MIN Li(Chinese Medicine School,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;Fujian Provincial 2011 Collaborative Innovation Center of TCM Health Management,Fuzhou,Fujian 350122,China;TCM Health Status Identification Center,Fuzhou Fujian 350122,China)

机构地区：[1]福建中医药大学中医学院,福建福州350122 [2]福建省中医健康管理2011协同创新中心,福建福州350122 [3]中医健康状态辨识研究室,福建福州350122

出　　处：《福建中医药》2023年第9期12-17,33,共7页Fujian Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81603541);福建省自然科学基金项目(2021J01131533)。

摘　　要：目的观察大柴胡汤加减方(MDD)对湿热证胆囊胆固醇结石(CS)小鼠胆固醇代谢基因SRBI、ABCG5/G8表达的影响。方法将24只雄性C57BL/6J小鼠按体质量随机分为空白组、模型组、中药组和西药组,每组6只。除空白组外,其余小鼠采用“2%高胆固醇致石饲料+内湿+外湿”的方法造模13周建立CS湿热证小鼠模型,通过观察胆囊形态、一般情况与旷场行为学实验评判CS湿热证模型是否建立成功。第14周开始,4组小鼠均按体质量0.02 mL/(g·d)灌胃干预,中药组和西药组分别给予1.17 g/mL大柴胡汤加减方药液和2.5 mg/mL熊去氧胆酸溶液,其余2组则给予生理盐水,共干预4周。ELISA法检测4组小鼠胆汁中总胆固醇(TC)含量、总胆汁酸(TBA)、磷脂(PL)的含量和血清中高密度脂蛋白胆固醇(HDL-C)含量、总胆固醇(TC)的含量,HE染色法观察肝脏细胞组织形态学变化,qPCR和Western blot分别检测B类清道夫受体Ⅰ(SRBI)、三磷酸腺苷结合盒转运体G5(ABCG5)、三磷酸腺苷结合盒转运体G8(ABCG8)mRNA与蛋白表达量。结果与空白组比较,模型组小鼠胆囊结石与湿热证候表现突出,肝脏脂肪样变性明显,胆汁中TC含量升高、TBA与PL含量均减少(P<0.05),血清中HDL-C含量降低、TC含量均增多(P<0.05),SRBI、ABCG5、ABCG8 mRNA相对表达水平均有明显上调(P<0.05),SRBI和ABCG8蛋白表达均量上调(P<0.05)。与模型组比较,中药组小鼠的肝脏脂肪变性程度减轻,胆汁中TBA、PL含量显著升高而TC含量均显著降低(P<0.05),血清中HDL-C升高、TC均降低(P<0.05),SRBI mRNA相对表达水平明显降低(P<0.05),ABCG8 mRNA及蛋白表达均呈明显下调(P<0.05)。结论SRBI、ABCG5/G8表达异常升高可能是CS湿热证的分子生物学机制之一。大柴胡汤加减方能通过抑制SRBI、ABCG5/G8的表达,调节胆固醇代谢,减少胆固醇过饱和胆汁的分泌,改善肝脏脂肪变性,发挥清热祛湿、利胆消石的作用,从而达到治疗湿热型胆石�Objective:To observe the effects of Modified Dachaihu Decoction(MDD)on the expression of cholesterol metabolism genes SRBI and ABCG5/G8 in cholesterol gallstone(CS)mice with damp-heat syndrome.Methods:A totel of 24 male C57BL/6J mice were randomly divided into blank group,model group,traditional Chinese medicine group and Western medicine group according to body weight,with 6 mice in each group.The method of"2%high cholesterol lithogenic feed+internal damp-heat+external damp-heat"was used to establish the model of CS damp-heat syndrome in mice for 13 weeks.By observing gallbladder morphology,general condition and open field behavior experiment,the success of CS damp-heat syndrome model was evaluated.From the 14th week,mice in the four groups were given a solution of 0.02 mL/g by gavage.The Chinese medicine and Western medicine groups respectively administered a solution of MDD at a concentration of 1.17 g/mL,and a solution of 2.5 mg/mL ursodeoxycholic acid.The remaining two groups were administered with 0.9%saline.The intervention lasted for a total of 4 weeks.The enzyme-linked immunosorbent assay(ELISA)method was used to measure the elevated levels of total cholesterol(TC),increased levels of total bile acids(TBA),phospholipids(PL)in the bile;decreased levels of high-density lipoprotein cholesterol(HDL-C)and elevated levels of total cholesterol(TC)in the serum of the four groups.Hepatic cellular histology changes were examined using the hematoxylin and eosin(HE)staining method.Furthermore,quantitative polymerase chain reaction(qPCR)and Western blot analysis were employed to determine the mRNA and protein expression levels of scavenger receptor Class-B type I(SRBI),ATP binding cassette transporter G5(ABCG5),and binding cassette transporter G8(ABCG8).Results:Compared with the blank group,cholecystolithiasis and damp-heat syndrome of mice in model group were more prominent,liver fatty degeneration was obvious,TC content in bile increased,TBA and PL contents decreased(P<0.05),serum HDL-C content decreased and TC con

关 键 词：胆囊胆固醇结石 湿热证 SRBI ABCG5/G8 大柴胡汤加减方 胆固醇代谢 

分 类 号：R285.5[医药卫生—中药学]