Based On The Imbalance Of Treg/Th17 Cells, This Paper Explores The Regulation Of Warm-clearing And Usage On The Intestinal Mucosal Inflammatory Response In Rats With Chronic And Subacute Liver Failure Intestinal Endotoxemia

Objective:To observe the effects of clearing temperature and combining method on the expression of endotoxin C-reactive protein,Treg-Th17 cells and the expression of inflammatory factors and transcription factors in the colon of rats with hepatic failure induced by enterogenous endotoxemia.To explore the mechanism of anti-hepatic failure by reducing endotoxemia,and to provide a new perspective and clinical basis for the treatment of Acute-on-chronic liver failure with traditional Chinese medicine.Methods:An experimental model of immune liver fibrosis was established by using bovine serum albumin(BSA).The model of IETM was established by intraperitoneal injection of D-Gal + LPS(combined with D-Gal 400 mg/kg,LPS 100 ?g/kg).The frequency of eg/Th17 cells,the expression of TNF-? TGF-?,IL-10,IL-17,IL-23,FoxP3,ROR-?t-associated protein and gene expression were observed.Results:1.Lipopolysaccharide:compared with the normal group,the level of endotoxin in the other groups increased at each time point(P<0.05),especially in the model group(P<0.01),and compared with the model group.The level of endotoxin in the positive control group decreased significantly at each time point(P>0.05),and the level of endotoxin in the other experimental groups decreased significantly(P<0.05).2.Procalcitonin and C-Reactive Protein:compared with the normal group,the levels of CRP in the other groups increased at each time point(P<0.05),especially in the model group(P<0.01),and compared with the model group.In the positive control group,there was a slight decrease in the level of CRP at each time point(P>0.05),and the level of CRP in the other experimental groups was lower than that in the control group(P<0.05),especially in the combination of clearing and warming group(P<0.05),and there was a significant difference between the two groups(P<0.05),and there was a significant difference between the two groups(P<0.05).Compared with the previous time point,the level of CRP in group PCT was significantly higher than that in control group(P<0.05).3.Frequency and ratio of Treg?Th17 cell:compared with the normal group,the frequency of TregsTh17 cells in the other groups increased at each time point(P<0.05),the most significant increase in the model group(P<0.01),and compared with the model group,the frequency of Th17 cells in the other groups was significantly higher than that in the model group(P<0.05).Tregt Th17 cells in the positive control group were slightly decreased at each time point(P>0.05),and the other experimental groups were all decreased significantly(P<0.05),especially in the combination of clearing and warming group(P<0.05).Compared with the normal group,the Treg/Thl7 ratio in the model group decreased significantly at each time point,and the cell loss occurred.Compared with the model group,the Treg/Th17 ratio of the two groups increased significantly at each time point(P<0.05),and gradually increased with time,the cell imbalance was corrected in the direction of Treg,the positive control group and the Qingfa group were corrected in the direction of Treg.There was no significant difference between the warm group and the control group(P>0.05).4.The levels of IL-10 TNF-? and IL--17AnIL-23 TGF-? in colon tissue:compared with the normal group,the expression of inflammatory factors in other groups increased significantly(P<0.05)(the increase of pro-inflammatory factor was significantly greater than that of anti-inflammatory factor,and the most significant increase was found in the model group(P<0.01).In the positive control group,the level of inflammatory factors decreased slightly(P>0.05),and the levels of inflammatory factors in the other experimental groups were decreased(P<0.05),especially in the combination of clearing and warming group(P<0.05).The inhibitory effect was obviously higher than that of anti-inflammatory factor.5.In vitro amplification of RT-PCR:compared with the normal group,the ROR-?t mRNA of FoxP3mRNA-ROR-?t in the other groups increased at each time point(P<0.05),and the most obvious increase was(P<0.01)in the model group,and compared with the model group,the ROR-?t mRNA in the other groups was higher than that in the model group.The positive control FoxP3mRNA-ROR-?t mRNA decreased slightly at each time point(P>0.05),and all the other experimental groups decreased P<0.05,especially in the combination of clearing and warming group(P<0.05),and increased in each experimental group compared with the previous time point(P<0.05).The decrease of ROR-?t mRNA in the combined method group was higher than that in the FoxP3mRNA group.6.In situ hybridization and immunohistochemistry of FoxP3 ROR-?t in colon tissue:compared with the normal group,the other groups of FoxP3,ROR-gamma t related protein and gene expression were increased(P<0.05)(ROR-gamma t related protein and gene elevation was greater than FoxP3),the model group increased most significantly(P<0.01),and the positive control group was slightly lower(P>0.05)compared with the model group.The expression of FoxP3 and ROR-?T gene and protein in all experimental groups decreased(P<0.05).The decrease of ROR-?t related protein and gene in the combined method group was higher than that in FoxP3 group.Conclusion:1.During the occurrence of IETM,Treg/Th17 cells were out of balance and shifted to Th17 direction,which resulted in related inflammation inhibition and increased release of pro-inflammatory factors,thus exacerbating the degree of enterogenous endotoxemia.2.Wenyang jiedu Huayu prescription can reduce the differentiation of Treg and Th17 cells by inhibiting the expression of Foxp3ROR-yt in enterogenic endotoxemia,and then alleviate the damage caused by the release of related proinflammatory and anti-inflammatory factors.Promote the balance of Treg/Th17 cells(anti-inflammatory/pro-inflammatory)to Treg recovery.