摘要
目的:探讨泽明红山方对大鼠代谢相关脂肪性肝病(MAFLD)合并混合型高脂血症(MHLP)模型的影响以及可能的机制。方法:SD大鼠给予高脂饲料8周建立MAFLD合并MHLP模型。随机分为模型组,中药低、高剂量组以及非诺贝特组;建立模型同时给予相应药物干预。对照组大鼠给予普通饲料。观察药物对模型大鼠血清学、肝脏病理、cAMP-PKA信号转导通路mRNA及脂肪酸转运相关蛋白表达的影响。结果:(1)体重及肝脏系数:各组内大鼠体重差异无统计学意义;模型组大鼠体重及肝脏系数显著升高,各干预组大鼠体重及肝脏系数均有不同程度下降,其中以高剂量组下降最为显著。(2)肝功能及血脂:与对照组比较,模型组大鼠的肝功能(ALT、AST)以及血脂水平(TG、TC、LDL-C、FFA)显著升高;与模型组比较,各治疗干预组大鼠肝功能及血脂水平均在不同程度得到改善。(3)肝脏病理染色:模型组大鼠肝小叶结构不清,胞浆内可见分布的脂滴空泡,伴大量炎性细胞浸润,证实肝脏表现出弥漫性脂肪变;各干预组大鼠肝细胞形态、肝组织结构呈现不同程度改善。(4)各组大鼠基因表达水平:与对照组比较,模型组大鼠cAMP、PKA及PPARα的基因表达水平显著下降,与模型组比较,各治疗干预组大鼠肝脏基因表达水平均在不同程度得到改善,其中高剂量组改善最为明显。(5)各组大鼠肝脏FABP1、CD36蛋白水平:与对照组比较,模型组大鼠肝脏中FABP1、CD36蛋白的表达显著升高,各干预组蛋白表达均有不同程度下调;其中高低剂量组大鼠肝组织FABP1、CD36蛋白改善效果显著。(6)各组大鼠肝脏组织cAMP、PPARα、CD36蛋白表达水平:与对照组比较,模型组大鼠肝组织中cAMP、PPARα蛋白染色浅,各干预组大鼠较模型组染色加深。与对照组比较,模型组大鼠肝组织中CD36蛋白染色深,各干预组较模型组染色浅。结论:泽明红山方可能通过cAMP-PKA通路,促进PPARα蛋白的激活,抑制FABP1和CD36的表达,并降低血清TG、TC、LDL-C、FFA水平,抑制肝脏摄取FFA过程,调节肝脏脂质代谢功能,缓解肝细胞炎症,逆转肝脏损伤,起到治疗MAFLD合并MHLP的作用。
Objective:Ze Ming Hong Shan prescription is an empirical formula for the clinical treatment of metabolic associated fatty liver disease,which has a positive effect on regulating liver function and blood lipids in MAFLD patients.This study explores the effects and possible mechanisms of this formula on a rat model of MAFLD combined with mixed hyperlipidemia.Methods:SD rats were given high-fat diet for 8 weeks to establish a MAFLD combined with MHLP model.They were Randomly divided into model group,low and high dose Chinese medicine group,and fenofibrate group.Establish a model and administer corresponding medication interventions.Give regular feed as the control group.Observe the effects of drugs on the serology,liver pathology,cAMP-PKA signaling pathway mRNA,and fatty acid transport related protein expression of model rats.Results:First,the body weight and liver coefficient:There was no statistically significant difference in body weight among rats in each group;The weight and liver coefficient of the model group significantly increased,and the weight and liver coefficient of rats in each intervention group decreased to varying degrees,with the high-dose group showing the most significant decrease.Secondly,the liver function and blood lipids:Compared with the control group,the liver function(ALT,AST)and blood lipid levels(TG,TC,LDL-C,FFA)of the model group rats were significantly increased;Compared with the model group,the liver function and blood lipid levels of rats in each treatment intervention group were improved to varying degrees.Thirdly,the liver pathological staining:the liver lobule structure of the model group rats was unclear,with lipid droplets and vacuoles visible in the cytoplasm,accompanied by a large number of inflammatory cell infiltration,confirming that the liver exhibited diffuse steatosis.The morphology of liver cells and liver tissue structure showed varying degrees of improvement in each intervention group.Fourthly,the effect on the mRNA expression of cAMP,PKA,PPARα,and FABP1 in liver tissue:Compared with the control group,the gene expression levels of cAMP,PKA,and PPARαin the model group rats were significantly reduced,while the gene expression level of FABP1 was significantly increased;Compared with the model group,the gene expression levels in the liver of rats in each treatment intervention group were improved to varying degrees,with the high-dose group showing the most significant improvement.Fifthly,The effect of Western blot on the expression of FABP1 and CD36 proteins in liver tissue was detected:compared with the control group,the expression of FABP1 and CD36 proteins in the liver of the model group rats was significantly increased,and the protein expression of each intervention group was downregulated to varying degrees;The improvement effect of FABP1 and CD36 proteins in liver tissue of rats in the high and low dose groups was significant.Sixthly,the immunohistochemical method was used to detect the effects of cAMP,PPARα,and CD36 protein expression in liver tissue:Compared with the control group,the cAMP and PPARαprotein staining in the liver tissue of the model group rats was lighter,while the staining in each intervention group was darker than that in the model group.Compared with the control group,the CD36 protein staining in the liver tissue of the model group rats was deeper,and the staining in each intervention group was lighter than that in the model group.Conclusion:Zequ Mingshan Formula may promote the activation of PPARαprotein,inhibit the expression of FABP1 and CD36,and lower serum levels of TG,TC,LDL-C,and FFA through the cAMP-PKA pathway.It can inhibit liver uptake of FFA,regulate liver lipid metabolism,alleviate liver cell inflammation,reverse liver damage,and play a role in treating MAFLD combined with MHLP.
作者
耿晓萱
卢秉久
郑佳连
GENG Xiao-xuan;LU Bing-jiu;ZHENG Jia-lian(Liaoning Affiliated Hospital of Traditional Chinese Medicine(Shenyang,110032),China;不详)
出处
《中西医结合肝病杂志》
2025年第5期564-570,共7页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
基金
全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号)
辽宁省科技厅省自然基金项目(No.2019-MS-231)
辽宁省教育厅科学技术研究项目(No.L201903)。
