健脾化浊饮对代谢相关脂肪性肝病大鼠肠肝轴体系影响的研究  

Study on the effect of Jianpi Huazhuo Yin on the gut-liver axis system in rats with metabolic-related fatty liver disease

作　　者：杨洪超 徐龙进[2] 孙建光[3] YANG Hongchao;XU Longjin;SUN Jianguang(Qingdao Hospital of Traditional Chinese Medicine(Haici Hospital),Qingdao 266000,Shandong,China;Shandong Center for Disease Control and Prevention,Jinan 250000,Shandong,China;Shandong Hospital of Traditional Chinese Medicine,Jinan 250000,Shandong,China)

机构地区：[1]青岛市中医医院(青岛市海慈医院),山东青岛266000 [2]山东省疾病预防控制中心,山东济南250000 [3]山东省中医院,山东济南250000

出　　处：《现代中西医结合杂志》2021年第24期2623-2630,2696,共9页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：山东省中医药科技发展计划项目(2009-49);山东省高等学校科技计划项目(J09LF14)。

摘　　要：目的研究健脾化浊饮对代谢相关脂肪性肝病(MAFLD)大鼠肠肝轴体系的影响,初步探讨健脾化浊饮的作用机制。方法随机选取8只健康SD大鼠作为正常组,予普通饲料喂养及生理盐水灌胃;将35只造模成功MAFLD大鼠随机分为模型组8只、健脾化浊饮低剂量组8只、健脾化浊饮中剂量组9只、健脾化浊饮高剂量组10只,均予以高脂饲料喂养,同时模型组灌胃生理盐水,健脾化浊饮低、中、高剂量组分别灌胃含生药10,15,20 g/kg的健脾化浊饮。灌胃12周后,检测各组大鼠血清总胆固醇(TC)、三酰甘油(TG)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转氨酶(AST)、空腹血糖(FPG)、脂多糖(LPS)、D-乳酸(D-Lac)水平;苏木素-伊红(HE)染色与油红O染色检测肝组织脂肪变性程度;回肠组织HE染色和肠道紧密连接蛋白Occludin免疫组化检测肠黏膜通透性;Western blot法检测肝组织中TLR4、NF-κB蛋白表达情况;RT-PCR法检测大鼠粪便中大肠杆菌、乳酸杆菌相对含量。结果与正常组比较,模型组大鼠血清ALT、AST、TC、TG、FPG、LPS、D-Lac水平及肝组织中TLR4、NF-κB蛋白表达量和粪便中大肠杆菌相对含量均明显升高(P均<0.05),肠组织中Occludin蛋白阳性染色颗粒明显减少,粪便中乳酸杆菌相对含量明显降低(P均<0.05);健脾化浊饮中、高剂量组大鼠血清ALT、AST、TC、TG、FPG、LPS、D-Lac水平及肝组织中TLR4、NF-κB蛋白表达量和粪便中大肠杆菌相对含量均明显低于模型组(P均<0.05),肠组织中Occludin蛋白阳性染色颗粒明显增多,粪便中乳酸杆菌相对含量明显高于模型组(P均<0.05)。结论健脾化浊饮对MAFLD大鼠肠肝轴体系具有调节作用,肠道细菌可能是其关键调节因子,其能通过调节肠道菌群,修复肠黏膜屏障,降低肠黏膜通透性,进而抑制LPS/TLR4/NF-κB信号通路,阻断肠肝损伤的恶性循环,从而达到治疗MAFLD的作用。Objective It is to study the effect of Jianpi Huazhuo Yin on the gut-liver axis system of rats with metabolic associated fatty liver disease,and to explore the mechanism of this decoction.Methods Eight healthy SD rats were randomly selected as the normal group,fed with normal feed and given normal saline by gavage;35 successfully modeled MAFLD rats were randomly divided into a model group(n=8),a low-dose Jianpi Huazhuo Yin group(n=8),a medium dose Jianpi Huazhuo Yin group(n=9)and a high dose Jianpi Huazhuo Yin group(n=10),all were fed with high-fat feed.At the same time,the model group was given normal saline by gavage,and the low,medium and high dose groups of Jianpi Huazhuo Yin were respectively given Jianpi Huazhuo Yin which contained 10,15,20 g/kg crude drugs by gavage.After intragastric administration for 12 weeks,the levels of total serum cholesterol(TC),triacylglycerol(TG),alanine aminotransferase(ALT),aspartate aminotransferase(AST),fasting blood glucose(FPG),lipopolysaccharide(LPS),D-lactic acid(D-Lac)of the rats in each group were measured;the steatosis of liver tissue was detected by hematoxylin-eosin(HE)staining and oil red O staining;the intestinal mucosal permeability was detected by HE staining of ileum tissue and immunohistochemistry of intestinal tight junction protein Occludin;the expression of TLR4 and NF-κB protein in liver tissue were detected by Western blot;and the relative content of Escherichia coli and Lactobacillus in rat feces were detected by RT-PCR method.Results Compared with the normal group,the levels of serum ALT,AST,TC,TG,FPG,LPS,D-Lac,the expression of TLR4 and NF-κB protein in liver tissue and the relative content of Escherichia coli in feces in the model group were significantly increased(all P<0.05),the positive staining particles of Occludin protein in the intestinal tissue were significantly reduced,and the relative content of Lactobacillus in the stool was significantly reduced(all P<0.05);the levels of serum ALT,AST,TC,TG,FPG,LPS,D-Lac,and the protein expression of TLR

关 键 词：代谢相关脂肪性肝病 肠肝轴 肠黏膜屏障 肠道菌群 健脾化浊饮 

分 类 号：R-332[医药卫生]