| Objective This study was aimed to observe the changes of plasma mitochondrial DNA level and lung tissue FPR activation and inflammatory injury in rats with SAP-ALI treated with Qingyi Decoction,and the protective mechanism of Qingyi Decoction by regulating DAMPs;And to detect the changes of microbial flora in rat feces by microbial 16 Sr DNA sequencing technology,and the effect of Qingyi Decoction by regulating intestinal microecology in rats.Method 1.Therapeutic mechanism of Qingyi Decoction regulating DAMPs on lung tissue and organ damage in SAP-ALI rats Thirty healthy male Wistar rats of SPF grade were divided into control,model and Qingyitang according to random number method.The Qingyi Decoction was given Qingyi Decoction 10 m L/kg one week before the experiment,and the other two groups were given the same amount of normal saline.The Qingyi Decoction and the model group were retrogradely injected with sodium taurocholate in the rat pancreatic duct.The control only gently flipped the pancreas after laparotomy,that is,the abdominal cavity was closed.After 24 hours,the rats were sacrificed by cervical dislocation.The specimens were collected and removed from the lung tissue,fixed in 10% formalin solution,partially stained with HE and IHC of FPR1.The pathological changes of lung tissue were observed under microscope and pathological damage was performed.Score,a portion of the homogenate was used to extract the supernatant of the tissue,and the levels of MPO,TNF-α,IL-1,and IL-6 were detected by ELISA,removing plasma to determine amylase and lipase levels,extracting free DNA from plasma,detecting circulating mt DNA content by RT-PCR,though Western blotting to detect each The protein content of FPR1 and FPR2 in the lung tissue of the rats,and the gene level by RTPCR.2.Using 16 Sr DNA sequencing technology to investigate the effect of Qingyi Decoction on the regulation of intestinal microflora in SAP rats The specimens were collected from colon and pancreas tissues in each group.The pathological changes of colon and pancreas were observed by HE.The cecal segment feces and 16 Sr DNA sequencing technique were used to analyze the changes of intestinal microflora in each group.Result Compared with the normal group,the level of mt DNA in circulating blood of SAP model rats was significantly increased（P<0.01）,the injury score of each organ was significantly increased（P<0.01）,and the levels of amylase and lipase,of MPO,TNFα,IL-1 and IL-6 in lung tissue were significantly increased（P<0.01）,and the relative expression of FPR1 and FPR2 m RNA and protein in lung tissue increased（P<0.01）,but Qingyi Decoction the group can effectively reduce these indicators（P <0.05）.The species difference between the three was statistically significant（P<0.05）.The results of three levels of analysis from the portal,the genus and the genus showed that the Qingyi Decoction group can effectively reduce the number of thick-walled bacteria including Clostridium,Bacillus,Ruminiclostridium9,Eubacterium and Helicobacter species in SAP rats.The content of the flora increased the content of Bacteroides including Bacteroides,Bacteroides and Lactobacillus（P<0.05）.Conclusion Qingyi Decoction can effectively reduce the mt DNA level in the circulating blood of SAP rats,and achieve the protection of the body organs,and the mt DNA in the circulating blood of SAP rats may be used as a judgment,and protecting the excessive inflammatory response of lung tissue in SAP-ALI by activating the expression of FPR1 and FPR2 on the surface of various inflammatory cells in lung tissue of SAP-ALI.And increasing the diversity and abundance of intestinal flora,regulating microbial ecological balance,increasing the content of probiotics,and reducing the colonization ability of harmful bacteria,thus achieving protection of the intestines. |
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