Research On The Influence Of Jiajiansanjiasan Which Based On The Theory "Zhukejiao" On The Balance Of Th17/Treg In Rats With Hepatic Fibrosis

Objective: To observe the effectiveness of Jiajiansanjiasan based on the theory of“Zhukejiao” in the treatment of hepatic fibrosis and the regulation of Th17/Treg balance.To reveal its mechanism from the perspective of immunology,and explore the theoretical basis of the theory of ‘Zhukejiao,enrich the scientific connotation of“Zhukejiao”.Method:1.set up immuno-hepatic fibrosis rat model by intraperitoneal injection of porcine serum,Jiajiansanjiasan intervention and Fufangbiejiaruanganpian as positive control drug,After 60 days of treatment,the liver function?hepatic fibrosis and blood rheology of rats was detected,observe the pathological morphology by HE staining and Masson staining to understand the degree of hepatic fibrosis in rats.2.To detect Th17 cell specific transcription factor ROR?t and Treg cell specific transcription factor Foxp3 gene expression level by the RT-PCR assay,The changes of ROR?t/ Foxp3 in each group were compared;ELISA method was used to detect the expression of Th17 cells related factors IL-6,IL-17,IL-21,IL-23 and Treg cell related factors IL-10?TGF-?1 in liver tissue,compare the change of Th17 and Treg cytokine ratio.Result:1.pathological observation There was no fibrosis change in normal group,in model group,obvious fibrous tissue hyperplasia in central vein and portal area,Jiajiansanjiasan group and biejiaruanganpian group fibrosis lower than model group;Jiajiansanjiasan group decreased significantly compared with the model group(p<0.01),significantly increased compared to normal group(p<0.01),had no significant difference comparing with biejiaruanganpian group(P > 0.05).2.liver function index Compared with the normal group,ALT and AST increased significantly in the rats of model group(p<0.01);compared with the model group,ALT,AST decreased significantly of Jiajiansanjiasan groups(p<0.05),with no significant difference between biejiaruanganpian group(P > 0.05).There was no significant difference in ALB and TP in each group(P > 0.05).3.liver fibrosis index The fibrosis index LN??-C? PC??HA of model group were significantly higher than that of the normal group(p<0.01);the fibrosis index LN ? PC ? ? HA in Jiajiansanjiasan group were lower than those in model group(p<0.01 or p<0.05),with no significant difference in ?-C(P > 0.05),with no significant difference in LN??-C? PC??HA between biejiaruanganpian group(P> 0.05).4.hemorheology index The model group's blood viscosity and RBC hematocrit in model group was significantly higher than the normal group(P < 0.01),with no significant difference in plasma viscosity(P> 0.05);Jiajiansanjiasan group's blood viscosity and RBC hematocrit were significantly lower than those in model group(p<0.05),in plasma viscosity(P >0.05);Compared with t biejiaruanganpian group,all showed no significant difference(P >0.05).5.Th17/Treg correlation detection Compared with the normal group,ROR?t?Foxp3?IL-10?IL-6?IL-17? IL-21 ?IL-23 in model group were increased significantly,ROR?t/ Foxp3,IL-6/TGF-?,IL-17/TGF-?,IL-6/ IL-10,IL-17/ IL-10 increased obviously(P < 0.01);ROR?t?Foxp3?IL-10?IL-6?IL-17?IL-21?IL-23 in Jiajiansanjiasan group were lower significantly than those in model group,ROR?t/Foxp3 ?IL-6/TGF-?,IL-17/TGF-?,IL-6/IL-10,IL-17/ IL-10 were lower significantly than those in model group(P < 0.01),Compared with biejiaruanganpian group,IL-17,IL-17/TGF-? in Jiajiansanjiasan group decreased significantly,IL-21 increased significantly(P<0.05).Conclusion:1.The rat model of hepatic fibrosis can be make successfully by intraperitoneal injection of porcine serum2.Jiajiansanjiasan can improve the general state of Hepatic fibrosis rats,improve the liver function,reduce the blood rheology index and the index of liver fibrosis,and reduce the degree of liver fibrosis.3.the expression of Th17 cell related factors in rats with hepatic fibrosis was dominant,suggesting that Th17/Treg balance was out of control,which was in favor of Th17.4.Jiajiansanjiasan can down regulate the levels of Th17 and Treg specific transcription factors and related cytokines in liver tissue of rat,made the balance of Th17 /Treg in the direction of Treg,which was beneficial to the control and reversion of liver fibrosis.