逍遥丸对代谢相关脂肪性肝炎大鼠LXR-α/SREBP-1c通路的调节机制研究  

Exploration on the regulation mechanism of LXR-α/SREBP-1c pathway in MASH rats based on the theory of"liver and large intestine communication"

作　　者：李梦琪 张馨月 张玉伟 孟雅楠 苗宇船 LI Meng-qi;ZHANG Xin-yue;ZHANG Yu-wei;MENG Ya-nan;MIAO Yu-chuan(Shanxi University of Chinese Medicine,Jinzhong,Shanxi 030619,China)

机构地区：[1]山西中医药大学,山西晋中030619

出　　处：《时珍国医国药》2024年第13期2919-2925,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(81470190);山西省卫生健康委项目(2019089);山西中医药大学科技创新能力培育计划“基础研究专项”项目(2021PY-JC-04);风湿免疫性疾病中西医结合基础研究(2024XKJS-03)。

摘　　要：目的探讨逍遥丸对代谢相关脂肪性肝炎(MASH)大鼠的部分治疗机制,阐明中医“肝与大肠相通”理论对治疗MASH的指导意义。方法选取成年雄性SD大鼠24只,将其随机分为空白组(Control组)8只,模型0组(Model 0组)16只。其中Control组给予普通饲料喂养,Model 0组给予高脂饮食喂养,40%四氯化碳背部皮下注射,同时给予饥饱失常和夹尾刺激,4周后,将Model 0组随机分为模型组(Model组)和逍遥丸组(XYW组),每组各8只。XYW组大鼠给予逍遥丸灌胃,其他两组给予生理盐水灌胃。给药4周后,分别测定大鼠肝功能和肝脂肪指标含量;苏木精-伊红(HE)染色观察肝组织病理变化;阿利新蓝-过碘酸雪夫(AB-PAS)染色观察肠道屏障受损情况;ELISA试剂盒测定大鼠肝匀浆中炎症因子水平;qRT-PCR测定大鼠肝脏LXR-α、SREBP-1c、Nrf2及结肠Claudin1、ZO-1、SREBP-1c、Nrf2 mRNA的表达;Western blot检测大鼠肝脏LXR-α、SREBP-1c、Nrf2及结肠Claudin1、ZO-1、SREBP-1c、Nrf2的蛋白表达情况。结果与Control组比较,Model组大鼠血清ALT、AST以及肝匀浆T-CHO、TG、LDL-C、IL-8、IL-17、TNF-α水平升高(P<0.05),HDL-C、IL-10、TGF-β1水平降低(P<0.01);Model组大鼠肝组织LXR-α、SREBP-1c mRNA表达升高(P<0.001),Nrf2mRNA表达降低(P<0.01),结肠组织Claudin1、ZO-1、Nrf2 mRNA的表达降低(P<0.01),SREBP-1c的表达升高(P<0.01);Model组大鼠肝组织LXR-α、SREBP-1c蛋白水平升高(P<0.01),Nrf2降低(P<0.05),结肠组织中Claudin1、ZO-1、Nrf2蛋白水平降低(P<0.001),SREBP-1c升高(P<0.001)。结论逍遥丸对MASH大鼠具有一定的治疗作用,其治疗机制可能与减轻炎症、氧化应激反应,抑制脂肪酸堆积有关;保护肠黏膜屏障免受损害可以有效减轻肝脏的损伤,“肝与大肠相通”理论对于MASH治疗具有一定的指导意义。Objective To explore the therapeutic mechanism of Xiaoyao pills on MASH rats,and to clarify the guiding significance of the theory of"connecting liver and large intestine"in TCM.Methods A total of 24 adult male SD rats were selected and randomly divided into Control group(8 rats)and Model group(16 rats).The Control group was fed with ordinary diet,and the Model 0 group was fed with high-fat diet,with subcutaneous injection of 40%carbon tetrachloride into the back,as well as with hunger and satiation disorder and tail-clip stimulation.4 weeks later,the Model 0 group was randomly divided into Model group and Xiaoyao pill group(XYW group),with 8 animals in each group.The rats in XYW group were given Xiaoyao pill and the other two groups were given normal sa⁃line.After 4 weeks of administration,the liver function and liver fat content of rats were measured respectively.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of liver tissue.The damage of intestinal barrier was observed by Alcian blue-periodate Shev(AB-PAS)staining.The levels of inflammatory factors in rat liver homogenate were determined by ELISA kit.The mRNA expressions of LXR-α,SREBP-1c and Nrf2 in liver and Claudin1,ZO-1,SREBP-1c and Nrf2 in colon were determined by qRT-PCR.The protein expression of LXR-α,SREBP-1c and Nrf2 in liver and Claudin1,ZO-1,SREBP-1c and Nrf2 in colon were detected by Western blot.Results Compared with Control group,the levels of serum ALT,AST and liver homogenate T-CHO,TG,LDL-C,IL-8,IL-17 and TNF-αin Model group were increased(P<0.05),while the levels of HDL-C,IL-10 and TGF-β1 were decreased(P<0.01).In Model group,the mRNA expressions of LXR-αand SREBP-1c in liver tissue were increased(P<0.001),the mRNA expressions of Nrf2 were decreased(P<0.01),and the mrna expressions of Claudin1,ZO-1 and Nrf2 in colon tissue were decreased(P<0.01).The expression of SREBP-1c was increased(P<0.01).The protein levels of LXR-αand SREBP-1c in liver tissue of Model group were increased(P<0.01),while Nrf2 was decreas

关 键 词：代谢相关脂肪性肝炎 逍遥丸 肝与大肠相通 肝X受体Α 固醇调节元件结合蛋白-1C 核因子E2相关因子 

分 类 号：R285.5[医药卫生—中药学]