大黄灵仙方调控外泌体LINC00311抑制胆道系统结石形成的影响  

Effect of Dahuang Lingxian prescription on inhibiting biliary stone formation by regulating exosome LINC00311

作　　者：黄欣[1,2] 俞渊[2] 张曼 武丽[1] 戴建业[1] 何晓微[1] HUANG Xin;YU Yuan;ZHANG Man;WU Li;DAI Jian-ye;HE Xiao-wei(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Guangxi University of Chinese Medicine,Nanning 530001,China)

机构地区：[1]广西中医药大学,广西南宁530001 [2]广西中医药大学第一附属医院,广西南宁530023

出　　处：《时珍国医国药》2024年第14期3171-3178,共8页Lishizhen Medicine and Materia Medica Research

基　　金：广西自然科学基金(2021GXNSFBA220059);广西中医药大学“岐黄工程”高层次人才团队培育项目(202411)。

摘　　要：目的探讨大黄灵仙方调控外泌体LINC00311抑制结石形成,防治胆石病的机制。方法70只SD雄性小鼠随机分为:空白对照组、模型组、熊去氧胆酸组、大黄灵仙组、模型+基因阻滞组(模型KO组)、熊去氧胆酸+基因阻滞组(熊去氧胆酸KO组)、大黄灵仙+基因阻滞组(大黄灵仙KO组),每组10只。采用高胆固醇、高热量致石饲料诱发的方式建立胆石病小鼠模型,再按体内转染实验法建立LINC00311基因敲除小鼠模型。连续干预6周后,记录各组小鼠成石率,电镜观察胆管组织内细胞超微结构的变化,RT-PCR检测LINC00311及TGF-β1/Smads信号通路中关键因子的表达,核磁共振氢谱技术检测胆汁内代谢产物表达情况。结果大黄灵仙组的成石率为0%,低于模型组、模型KO组、大黄灵仙KO组(P<0.002)。电镜检测显示大黄灵仙组细胞器形态结构完整,优于模型组、模型KO组、大黄灵仙KO组。RT-PCR显示与模型组、模型KO组、大黄灵仙KO组相比,大黄灵仙组的LINC00311、Smad4、Smad7升高(P<0.05),TGF-β1、Smad2、Smad3降低(P<0.05)。核磁共振氢谱检测显示与模型组相比,大黄灵仙组胆汁中有9种代谢产物具有明显的差异性。结论大黄灵仙方能促进外泌体LINC00311在胆道系统内的聚集,改善细胞结构,调节炎症反应和纤维化,稳定胆汁代谢,具有防治胆石病的作用。Objective To investigate the mechanism of Dahuang Lingxian prescription in inhibiting stone formation and preventing cholelithiasis by regulating exosome LINC00311.Methods The 70 male SD mice were randomly divided into blank control group,model group,ursodeoxycholic acid group,Dahuang Lingxian group,model+gene block group(model KO group),ursodeoxycholic acid+gene block group(ursodeoxycholic acid KO group),Dahuang Lingxian+gene block group(Dahuang Lingxian KO group),with 10 mice in each group.The mice model of cholelithiasis was induced by high cholesterol and high calorie lithogenic diet.The LINC00311 gene knockout mice model was established according to the in vivo transfection experiment.After 6 weeks of continuous intervention,the stone formation rate of mice in each group was recorded,the ultrastructural changes of cells in bile duct tissue were observed by electron microscope,the expressions of LINC00311 and TGF-β1/Smads signaling pathway were detected by RT-PCR,and the expression of metabolites in bile was detected by 1H NMR.Results The stone formation rate of the Dahuang Lingxian group was 0%,which was lower than model group,model KO group,and Dahuang Lingxian KO group(P<0.002).Electron microscopy showed that the structure of organelles in the Dahuang Lingxian group were complete,which was better than that in the model group,model KO group,and Dahuang Lingxian KO group.RT-PCR showed that LINC00311,Smad4 and Smad7 in Dahuang Lingxian group were significantly higher than those in model group,model KO group and Dahuang Lingxian KO group(P<0.05),while TGF-β1,Smad2 and Smad3 were decreased(P<0.05).1H NMR showed that there were 9 metabolites in the bile of Dahuang Lingxian group with significant differences compared with model group.Conclusion Dahuang Lingxian prescription can promote the accumulation of exosome LINC00311 in the biliary system,improve cell structure,regulate inflammatory response and fibrosis,stabilize bile metabolism,prevent and treat cholelithiasis.

关 键 词：胆石病 大黄灵仙方 外泌体LINC00311 信号传导 胆汁代谢 

分 类 号：R285.5[医药卫生—中药学]