| Objective: The 16 S r DNA sequencing technology was used to observe the distribution characteristics of intestinal flora of spontaneously hypertensive rats fed with high-fat diets,and to explore the effect of huanglianjiedu decoction on intestinal flora of the model rats,so as to provide a theoretical basis for the prevention and treatment of hypertension with intestinal flora as the target of traditional Chinese medicine.Methods: 1.Experimental grouping: 20 12-week-old male spf-rated WKY rats and 30 SHR rats were randomly divided into the normal control group(group A,n=10)and the normal high-fat group(group B,n=10)for adaptive feeding for 1 week.SHR was randomly divided into the hypertensive control group(group C,n=10).20 rats were fed with normal diet and the model group were fed with high-fat diet.All rats were raised continuously for8 weeks.At the end of 8 weeks,the model group was randomly divided into the hypertensive high-fat group(group D,n=10)and the huanglian jiedu decoction group(group H,n=10).The rats’ diets remained unchanged.2.Test and analysis: the systolic blood pressure of rats in each group was measured by non-invasive blood pressure tester every week.After 12 weeks,the rats fasting,the water 8 h,press 3 ml/kg intraperitoneal injection of 10%chloral hydrate anesthesia,about 2 cm from the cecum and end intercept in the rat cecum,remove the cecum contents(fresh feces),tag placement after60 ml box and immediately frozen in the ice,and then transferred to-80 ℃refrigerator aside,entrust Shanghai beauty,biomedical technology co.,LTD.(feces)in the cecum and the sample of 16 s r DNA detection.PE reads obtained by Miseq sequencing were firstly splicing according to overlap relationship,and sequence quality was controlled and filtered at the same time.After samples were separated,OTU clustering analysis and species taxonomy analysis were carried out,and OTU Venn diagram and PCo A diagram were used for display.The microflora differences and the changes of microflora structure at the level of phylum,class,family and genus of intestinal microflora among each group were further analyzed,which were displayed by colony Bar map and colony Heatmap.Results: 1.Blood pressure level of rats in each group: before the experiment,there was no statistically significant difference in blood pressure between rats of the same species.In WKY rats,compared with group A of the same period,blood pressure in group B increased at week 8 and 12,with statistically significant difference(P<0.05).In SHR,compared with group C of the same period,blood pressure in group D and group H increased significantly at week 8,with statistically significant difference(P<0.05).At12 weeks,the blood pressure of group D was significantly higher than that of group C and group H,and the difference was statistically significant(P <0.05).2.SHR structure of intestinal flora characteristics: compared with group A,group C significantly reduce the number of OTUs,analysis of PCo A PC1 dimension and PC2 had no significant difference,A significant reduction in the abundance of intestinal flora,no difference between the diversity and flora structure difference of deformation of thick wall bacteria,bacteria,actinomyces,green turn bacterium,lactobacillus decreased significantly,rumen bacteria,la bacteria bacterium such as increased significantly.3.Effect of high-fat diet on intestinal microflora of WKY rats: compared with group A,the number of OTUs in group B decreased,and the abundance of intestinal microflora showed A significant decline.In PCo A analysis,significant differences and remote clustering distances were found in PC1 dimension and PC2 dimension.Intestinal flora showed increased abundance of firmicutes,roscorella,bratistia and clostridium.The abundance of bacteroidetes,lactobacillus,prevotella and rumen cocci decreased.4.Effect of high-fat diet on SHR intestinal flora: compared with group C,the number of OTUs in group D decreased,and the abundance of intestinal flora showed a significant decline.In PCo A analysis,significant differences and remote clustering distances were found in PC1 dimension and PC2 dimension.Intestinal flora showed increased abundance of firmicutes,clostridium,helicobacter,veroniella,monomonas and bratia.The abundance of bacteroides,spirochetes,bacillus,bacteroides,lactobacillus,prevotella,and rumen coccus decreased.5.Effect of rhizoptidis decoction on intestinal flora of high-fat fed SHR:compared with group D,the number of OTUs in group H increased,and the abundance of bacteroides,bacilli and lactobacillus increased.The abundance of firmicutes,helicoids and megalomonas decreased significantly.Conclusion(s): 1.The abundance of intestinal microorganisms in SHR was significantly lower than that in WKY rats,and the intestinal flora in hypertension was in a state of disorder.2.The high-fat diet significantly changed the intestinal flora structure of WKY rats and raised blood pressure,which may be related to the significantly reduced abundance of probiotics such as lactobacillus and the significantly increased abundance of conditional pathogenic bacteria such as firmicutes.3.The effect of high-fat diet on SHR intestinal flora was similar to that of WKY rats,but the colonization resistance of intestinal flora was destroyed,and the specific expression was the disappearance of spirochete and the emergence of new strain megamonas.4.Huanglian Jiedu Decoction can effectively improve the disorder of intestinal flora,repair intestinal colonization resistance,increase the abundance of flora and probiotics,reduce the abundance of pathogenic bacteria and achieve the effect of anti hypertension. |
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