Effect Of Yiqi Huayu Qingre Formula And Its Compositions On MTOR Signaling Pathway In Rats With IgA Nephropathy

Objective: To observe the Yiqi Huayu Qingre formula and its compositions’ effect on experimental rats model with IgA nephropathy,in the experiment of each group rats’ nephridial tissue expression,we will observe the phosphorylation expression of downstream signaling protein S6 and 4EBP1 that of mTOR signaling pathway.And then further investigate the mechanism of mTOR signaling pathway and Yiqi Huayu Qingre formula and its compositions’ curative effect on IgA nephropathyMethods: Wistar male rats of SPF grade were randomized allocation to 6 groups:blank group,model group,Yiqi group,Huayu group,Qingre group,herb compound group.Using bovine serum albumin(BSA)as antigen filling the stomach for every two days to stimulate the rats,injecting carbon tetrachloride(CCl4)to rats ’ neck skin weekly,injecting lipopolysaccharide as immune adjuvant at the 6th week through tail vein to make IgA nephropathy model.In this experiment,we will observe the expression location of downstream signaling protein S6 and 4EBP1 that of mTOR signaling pathway by using the technology of immunohistochemistry,and then give them semi-quantitative analysis.Then we use the technology of Western blot to observe the quantity of renal cortex p S6 and p-4EBP1 expression that in each group of rats,then further investigate the mechanism of Yiqi Huayu Qingre formula and its compositions.Results:1 In the expression of p S6 protein1.1 The phosphorylation protein S6 which in the nephridial tissue of rats model with IgA nephropathy is mainly expressed in glomerular podocyte and epithelial cells of parietal layer,and sporadically expressed in renal tubules.1.2 As the result of immunohistochemistry,in all experimental groups of the quantity of protein p S6,the model group is maximum,and the Yiqi group takes the second place,and the order of the remaining groups is: Huayu group,Qingre group,herb compound group and blank group,at the same time,of these experimental groups,the differences between Huayu group and herb compound group,Qingre group and herb compound are meaningless in statistics,but the differences between each experimental group and model group are meaningful in statistics.1.3 As the result of Western blot,in all experimental groups of the quantity of phosphorylation protein p S6,the model group is maximum,and the Yiqi group takes the second place,and the Huayu group,Qingre group and herb compound group are minimum.What’s more,the result of Western blot group is consistent with the immunohistochemistry group.2 In the expression of protein p-4EBP12.1 The phosphorylation protein 4EBP1 which in the nephridial tissue of rats model with IgA nephropathy is mainly expressed in the glomerular podocyte and epithelial cells of parietal layer which in renal cortex,and sporadically expressed in renal tubules.2.2 As the result of immunohistochemistry,in all experimental groups of the quantity of protein p-4EBP1,the model group is maximum,and the Yiqi group,Huayu group and Qingre group take the second place,and the herb compound group and blank group are minimum,at the same time,of these experimental groups,the differences among Yiqi group,Huayu group and Qingre group are meaningless in statistic,but the differences between each experimental group and model group are meaningful in statistics,the blank group also have the sporadic expression.2.3 As the result of Western blot,in all experimental groups of the quantity of p-4EBP1,the model group is maximum,and the order of the remaining groups is:Yiqi group,Huayu group and Qingre group,and the herb compound group and blank group are minimum,at the same time,of these experimental groups,the differences among Yiqi group,Huayu group and Qingre group are meaningless in statistic(P > 0.5),but the differences between each experimental group and model group are meaningful in statistics.What’s more,the result of Western blot group is consistent with the immunohistochemistry group.Conclusion: According to the experiment above,we can know that:(1)MTOR signal pathway was abnormal activation in IGAN;(2)The Yiqi Huayu Qingre formula can play a therapeutic effect by inhibiting the abnormal activation of mTOR.The effect of herb compound group is the strongest,the Qingre group and Huayu group take the second place,and the Yiqi group has the minimum effect;(3)p S6 and p-4EBP1,the downstream effect protein of mTOR signaling pathway,express themself not only in the renal tubules but also in the glomerular podocytes and wall epithelial cells in IGAN rat model.