| Objective:To observe the effects of Wenyang Jiedu Huayu prescription and stasis on the imbalance of Treg/Th17 cells in rats with endotoxemia,and to explore the mechanism of anti-liver failure.Methods:160 SD male rats,random ceding 10 for the normal group,the remaining 150 sensitization to bovine serum albumin injection is adopted to establish the immune liver fibrosis in rats model,and then the rat model of liver failure was established by intraperitoneal injection of D-Gal+LPS solution,the normal group in the isodose physiological saline injection.It was randomly divided into the model group,the positive control group,the Qingfa group,the Wenfa group,and the Qingwen Bingyongfa group.In the first week of the acute attack,distilled water,fructose solution,Yenchenhao decoction,Yinchen Zhufu decoction and Wenyang Jiedu Huayu prescription were administered to irritate the stomach,and the normal group was given distilled water to irritate the stomach.Observed between groups of rats with acute attack 1h,12h,24h after liver function,Intravenous endotoxin and liver pathological morphological changes,and to detect peripheral blood Treg and Th17 cells proportion and Treg/Th17 ratio,liver tissue IL-10,TGF-beta,IL-17 a,IL-23,the level of TNF-a and FoxP3,ROR-?t mRNA expression.Results:1.Liver function:compared with normal group,ALT and TBIL in the model group increased significantly at each time point(P<0.01).Compared with the model group,ALT and TBIL of the positive control group and each experimental group decreased at each time point(P<0.05).Compared with positive control group,each group ALT,TBIL in various time points were lower(P<0.05),of which the Qingwen Bingyongfa group reduce the obvious(compared with the Qingfa group,the Wenfa group,P<0.05).2.Intravenous endotoxin:compared with the normal group,the endotoxin level of the model group was significantly increased at each time point(P<0.01).Compared with the model group,the difference in the positive control group was not statistically significant(P>0.05),and the endotoxin of each experimental group decreased at all points(P<0.05).Compared with positive control group,each group of endotoxin levels at various time points were lower(P<0.05),of which the Qingwen Bingyongfa group reduce the obvious(compared with the Qingfa group,the Wenfa group,P<0.05).3.Hepatic histopathology:compared with the normal group,liver tissue injury in the model group was significantly increased,and the pathological score was significantly increased at each time point(P<0.01).Compared with the model group,the liver tissue injury of the positive control group and the experimental group was reduced,and the pathological score decreased at all time points(P<0.05).Compared with the positive control group,the liver tissue injury of each experimental group was reduced,and the inflammatory response was the lowest in the Qingwen Bingyongfa group.4.The ratio of Treg and Th17 cells in peripheral blood and Treg/Th17 ratio:compared with the normal group,the Treg and Th17 of the model group were significantly increased at each time point(P<0.01),and Th17 was higher than Treg,and the Treg/Th17 ratio decreased significantly.Compared with the model group,the difference in the positive control group was not statistically significant(P>0.05),and the Treg and Th17 of each experimental group decreased at all points(P<0.05).Compared with positive control group,each group Treg and Th17 at various time points were lower(P<0.05),and the trend of Th17 lower than Treg,in which the Qingwen Bingyongfa group reduce the obvious,marked increase of the ratio of Th17/Treg(compared with the Qingfa group,the Wenfa group,P<0.05).5.Hepatic tissue il-10,TNF-alpha and il-17a,il-23,and TGF-beta levels:compared with the normal group,the anti-inflammatory and pro-inflammatory factors of the model group were significantly increased at each time point(P<0.01),and the pro-inflammatory factors were higher than the anti-inflammatory factors.Compared with the model group,the difference in the positive control group was not statistically significant(P>0.05),and the anti-inflammatory and pro-inflammatory factors of each experimental group decreased at all points of time(P<0.05).Compared with positive control group,each group anti-inflammatory and proinflammatory factor at various time points were lower(P<0.05),and the trend of proinflammatory factor lower than anti-inflammatory factors,of which the Qingwen Bingyongfa group reduce the obvious(compared with the Qingfa group,the Wenfa group,P<0.05).6.Hepatic tissue Foxp3 and ROR-yt mRNA expression:compared with the normal group,Foxp3 and ROR-yt in the model group were significantly increased at each time point(P<0.01),and ROR-yt was higher than Foxp3.Compared with the model group,the positive control difference was not statistically significant(P>0.05),and Foxp3 and ROR-yt in each experimental group decreased at all points(P<0.05).Compared with positive control group,each group Foxp3,ROR-yt at various time points were lower(P<0.05),and the trend of ROR-yt lower than Foxp3,of which the Qingwen Bingyongfa group reduce the obvious(compared with the Qingfa group,the Wenfa group,P<0.05).Conclusion:1.In the condition of hepatic failure,Treg/Th17 cells are out of balance,Th17 proportion is higher than Treg,and the imbalance is skewed towards Th17,leading to uncontrolled inflammation and severe necrosis of hepatocytes,which may be one of the pathologic mechanisms of liver failure.2.Wenyang Jiedu Huayu prescription can reverse the Treg and Thl 7 at the same time,lower the Th17 trend higher than that of Treg,promote Treg/Th17 cells to restore balance,in order to reduce the body's immune injury,inhibiting liver inflammation,promote liver cell damage repair,may be one of the mechanism of action of the party anti-liver failure. |
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