基于肠道菌群探讨坤安煎剂对去卵巢大鼠脂代谢的影响

Effects of Kun′an Decoction on Lipid Metabolism in Ovariectomized Rats from Perspective of Intestinal Flora

目的:基于肠道菌群变化探讨坤安煎剂对去卵巢大鼠脂代谢的影响。方法:从40只大鼠中随机选取10只作为空白组,其余大鼠采用双侧卵巢摘除法建立围绝经期模型。将造模成功的大鼠随机分为模型组、坤安煎剂组及雌激素组,连续灌胃10周。HE染色观察肝脏、腹主动脉病理形态,ELISA法检测血清卵泡刺激素(follicle stimulating hormone,FSH)、雌二醇(estradiol,E 2)含量,全自动生化分析仪检测血清总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate transaminase,AST)含量,宏基因组测序技术检测肠道菌群。结果:与空白组比较,模型组大鼠血清FSH、TC、TG、ALT、AST含量升高,E 2含量降低(P<0.01)。与模型组比较,坤安煎剂组大鼠血清FSH、TC、LDL-C、ALT、AST含量降低,E 2、HDL-C含量升高(P<0.01)。HE染色显示,空白组大鼠肝细胞和腹主动脉形态无明显异常。模型组大鼠的肝组织脂肪变性,伴细胞坏死;腹主动脉细胞排列不规则,内膜增厚。与模型组比较,坤安煎剂组和雌激素组大鼠肝细胞和腹主动脉病理形态明显改善。宏基因组测序显示,在种水平上,坤安煎剂组大鼠Limosilactobacillus reuteri、Lactobacillus johnsonii丰度较模型组升高(P<0.05)。物种差异性分析显示,空白组大鼠的特征微生物包括拟杆菌科(Bacteroidaceae)、居海事城球杆菌属(Phocaeicola)及埃氏拟杆菌(Bacteroides eggerthi)等。模型组大鼠的特征微生物包括联合乳杆菌属(Ligilactobacillus)、鼠联合乳杆菌(Ligilactobacillus murinus)、动物联合乳杆菌(Ligilactobacillus animalis)等。坤安煎剂组大鼠的特征微生物包括黏液乳杆菌属(Limosilactobacillus)、罗伊黏液乳杆菌(Limosilactobacillus reuteri)、粪普雷沃菌(Prevotella copri)等。与模型组比较,坤安煎剂组大鼠D-丙氨酸代谢、D-谷氨酰胺和D-谷氨酸代谢通路的丰度升高,缬氨酸、亮氨酸和异亮氨酸的生物合成通路的丰度降低(P<0.05)。Spearman相关性分析显示,大鼠血脂(LDL-C、HDL-C、TC、TG)水平与Lactobacillus johnsonii、Limosilactobacillus reuteri丰度存在相关性。结论:坤安煎剂可能通过调节肠道菌群从而发挥改善去卵巢大鼠脂代谢紊乱的作用。

Objective:To explore the effects of Kun′an Decoction on lipid metabolism in ovariectomized rats based on changes in intestinal flora.Methods:Ten rats were randomly selected from 40 rats as the blank group,and the rest of the rats were modeled as perimenopausal by bilateral ovary removal method.The rats were randomly divided into the model group,the Kunan decoction group and the estrogen group,and were gavaged continuously for 10 weeks.HE staining was used to observe the pathological morphology of the liver and the abdominal aorta,and serum follicle stimulating hormone(FSH)and estradiol(E 2)were detected by ELISA,and serum total cholesterol(tcal)was detected by fully automated biochemistry analyzer.Serum total cholesterol(TG),triacylglyceride(TC),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C)was detected.lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),alanine aminotransferase(ALT),and aspartate transaminase(AST),and intestinal flora were detected by macro-genome sequencing technology.Results:Compared with the blank group,the serum FSH,TC,TG,ALT,AST content of rats in the model group increased,and the E 2 content decreased(P<0.01).Compared with the model group,the serum FSH,TG,LDL-C,ALT,AST content of rats in Kun′an Decoction group decreased,and E 2,HDL-C content increased(P<0.01).HE staining showed that there was no obvious abnormality in the morphology of hepatocytes and abdominal aorta of the rats in the blank group.In the model group,fatty degeneration of liver tissue was accompanied by cell necrosis;the cells of the abdominal aorta were irregularly arranged,and the inner membrane was thickened.Compared with the model group,the pathologic morphology of hepatocytes and abdominal aorta in the Kun′an Decoction and estrogen groups improved significantly.Macrogenomic sequencing showed that at the species level,the abundance of Limosilactobacillus reuteri and Lactobacillus johnsonii was elevated in the Kun′an Decoction group compared with that of the model group(P<0.05).Species differentiation analysis showed that the characteristic microorganisms of rats in the blank group included Bacteroidaceae,Phocaeicola,and Bacteroides eggerthi,etc.In the model group,the abundance of Limosilactobacillus reuteri and Lactobacillus johnsonii was higher than that in the model group(P<0.05).Characteristic microorganisms of rats in the model group included Ligilactobacillus,Ligilactobacillus murinus and Ligilactobacillus animalis.Characteristic microorganisms of rats in the Kun′an Decoction group included Limosilactobacillus,Limosilactobacillus reuteri and Prevotella copri.Compared with the model group,the abundance of D-alanine metabolism,D-glutamine and D-glutamate metabolism pathway increased,and the abundance of valine,leucine and isoleucine biosynthesis pathway decreased in the kunan decoction group(P<0.05).With Spearman′s correlation analysis it is found that the levels of blood lipids(LDL-C,HDL-C,TC,TG)and Lactobacillus johnsonii were significantly different from each other in the rats.Lactobacillus johnsonii,Limosilactobacillus reuteri abundance were correlated.Conclusion:Kun′an Decoction may play a role in improving lipid metabolism disorders in ovariectomized rats by regulating intestinal flora.