| Objective:A rat model of sepsis was established by injecting lipopolysaccharide(LPS)into the tail vein to investigate the effect of Huayu Decoction on inflammatory factors in septic rats and myocardial protection in septic rats,and further detection of phlegm To investigate the expression of TLR4 signaling pathway protein and miRNA-146 a in septic rats,and to explore the mechanism of Huayu Decoction on myocardial protection in septic rats,and provide new ideas and methods for clinical treatment of sepsis.Methods:Experiment 1: 24 healthy male Wistar rats were randomly divided into: blank group,model group and Huayu prescription Chinese medicine group,8 in each group.The blank group: rats were pre-administered with normal saline for 1 week,once a day,and given intravenous saline to the tail vein on the 8th day;model group: rats were pre-administered with normal saline for 1 week,once a day,and at 8th In the last day,LPS was administered to the tail vein for sepsis modeling;Chinese medicine group: rats were pre-administered with phlegm and resolving phlegm and blood stasis for 1 week,once a day,and on the 8th day,LPS was administered to the tail vein for sepsis modeling.The general condition of the rats after modeling was observed and recorded.After 6 hours of modeling,the levels of serum inflammatory factors TNF-α and IL-1were detected by ELISA.The myocardial sections of the rats were stained with HE and the pathological changes of myocardial tissue were observed under microscope.Experiment 2: 32 Thirty-two healthy male Wistar rats were randomly divided into miRNA-146 a negative control group,miRNA-146 a negative agent + Chinese medicine group,miRNA-146 a inhibitor group,miRNA-146 a inhibitor + Chinese medicine group.miRNA-146 a negative control group: rats were pre-administered with normal saline for1 week,once a day,on the 8th day,LPS was administered to the tail vein for sepsis modeling.The tail vein was injected with miRNA-146 a antagomir 24 h before modeling.Nc;miRNA-146 a negative agent + Chinese medicine group: rats pre-administered Chinese medicine for 1 week,once a day,on the 8th day,LPS was administered to the tail vein for sepsis modeling,and tail vein injection of miRNA-146 a was given 24 hours before modeling.Antagomir-nc;miRNA-146 a inhibitor group: rats were pre-administered with normal saline for 1 week,once a day,on the 8th day,LPS was injected into the tail vein for modeling,and tail vein injection of miRNA-146 a antagomir was performed 24 hours before modeling.miRNA-146 a low expression treatment;miRNA-146 a inhibitor + Chinese medicine group: rats pre-administered Chinese medicine for 1 week,once a day,on the 8th day,LPS was administered to the tail vein for sepsis modeling,and the tail was given 24 hours before modeling.Low expression of miRNA-146 a was performed by intravenous injection of miRNA-146 a antagomir.The expression of miRNA-146 a in serum of each group was detected by RT-PCR.The levels of serum inflammatory factors TNF-α and IL-1 were detected by ELISA.Myocardial tissue was detected by Western Blot.TLR4,NF-κB protein expression,myocardial tissue HE staining,microscopic observation of myocardial histopathological changes.Results:Experiment 1:1.General observation after animal modeling: Compared with the rats in the blank group,the rats in the model group and the rats in the Chinese medicine group showed loss of appetite,reduced drinking water,mental wilting,and decreased reaction activity,but the rats in the traditional Chinese medicine group Psychoactive activity is better than model group.2.Pathological changes of myocardial tissue in each group: The myocardial cells in the sepsis model group were swollen and ruptured,the nucleus overflowed and ruptured,the myocardial fibers were broken,and the inflammatory cells invaded.The myocardial injury in the Chinese medicine group was reduced compared with the model group,and the myocardial injury was improved,but there was still edema and inflammatory cell infiltration,and some myocardial cells ruptured.3.The results of TNF-α and IL-1 in the rats of each group showed that compared with the blank group,the levels of serum inflammatory factors TNF-α and IL-1 in the Chinese medicine group and the model group were significantly increased,and the difference was statistically significant.(P<0.01);Compared with the model group,the levels of serum inflammatory factors TNF-α and IL-1 in the Chinese medicine group were significantly lower,and the difference was statistically significant(P<0.01).Experiment 2:1.General observation after animal modeling: The rats in the four groups all had loss of appetite,reduced drinking water,mental wilting,and yellowish vertical hair color,but the miRNA-146 a inhibitor + Chinese medicine group was superior to miRNA.Rats with-146 a inhibitor,miRNA-146 a negative agent + Chinese medicine group were superior to miRNA-146 a negative control group rats.2.Pathological changes of myocardial tissue in each group: The myocardial cells of each group showed incomplete structure,nucleus rupture and necrosis,myocardial fiber rupture,irregular alignment,inflammatory factor infiltration,tissue congestion and edema.Myocardial injury was the most severe in the miRNA-146 a inhibitor group.Myocardial injury in the miRNA-146 a inhibitor + Chinese medicine group was relieved compared with the miRNA-146 a inhibitor group,and myocardial damage was improved.3.The serum miRNA-146 a,TNF-α,and IL-1 levels in the rats in each group showed that the expression of miRNA-146 a was decreased in the miRNA-146 a inhibitor group compared with the miRNA-146 a negative control group.Significantly statistically significant(P<0.01),miRNA-146 a negative agent + Chinese medicine group miRNA-146 a expression increased(P <0.01);compared with miRNA-146 a inhibitor group,miRNA-146 a inhibitor The expression level of miRNA-146 a in the Chinese medicine group was increased(P<0.01).Compared with the miRNA-146 a negative control group,the serum inflammatory factors TNF-α and IL-1 in the miRNA-146 a inhibitor group.The level of serum inflammatory factors TNF-α and IL-1 was decreased in miRNA-146 a negative agent + Chinese medicine group(P<0.01);compared with miRNA-146 a inhibitor group The levels of serum inflammatory factors TNF-α and IL-1in miRNA-146 a inhibitor + Chinese medicine group were decreased(P<0.01).4.The results of TLR4 and NF-κB protein in myocardial tissue of rats in each group showed that the levels of TLR4 and NF-κB in myocardial tissue of rats in miRNA-146 a inhibitor group were higher than those in miRNA-146 a negative control group.(P<0.01),the levels of TLR4 and NF-κB in myocardial tissue of miRNA-146 a negative agent+Chinese medicine group were decreased(P<0.01).Compared with miRNA-146 a inhibitor group,miRNA-146 a inhibitor+ The levels of TLR4 and NF-κB in myocardial tissue of rats in the Chinese medicine group were decreased(P<0.01).Conclusions:1.Huayu Recipe Chinese medicine can reduce the release of inflammatory factors TNF-αand IL-1 in septic rats,reduce the inflammatory response in rats with sepsis,and reduce the myocardial damage in septic rats.2.miRNA-146 a can negatively regulate TLR4 signaling pathway in sepsis.When miRNA-146 a is inhibited,it can increase the expression of TLR4 signaling pathway protein and increase the levels of inflammatory factors TNF-αand IL-1,resulting in inflammatory response.Intensified,leading to further exacerbation of myocardial damage in septic rats.3.Huayu Recipe further up-regulates TLR4 signaling pathway by up-regulating miRNA-146 a expression,reduces TLR4 and NF-κB protein expression in signaling pathways,and reduces the release of inflammatory factors TNF-αand IL-1,reducing sepsis It brings excessive inflammatory reaction and has protective effect on myocardium of septic rats. |
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