Effects Of Tangshen Heji On The Expression Of SOCS-1, TGF-β1 And VEGF In Kideny Of Diabetic Nephropathy Rats

Background and purposeDiabetic nephropathy is the most common chronic complications of diabetes, and there is an upward trend in the incidence of a disease. The pathogenesis of DN involves a variety of factors such as the hemodynamic abnormalities, genetic, oxidative stress state of chronic inflammation, metabolic disorders, and cell signaling pathways. Among them, the inflammation factor caused by the activation of multiple cell signaling pathways play an important role, in recent years, studies have shown that Janus kinase/signal transduction and transcriptional activation factor (JAK/STAT) signaling pathway activation can cause inflammation of the kidney tissues cascade amplification reaction, is the regulation of gene expression of inflammation[1-2]. In some cases, the JAK/STAT pathway can be excessive activation,it can give a series of target genes including transforming growth factor-betal (TGF-beta 1)abnormal active biological function,and lead to proliferation, inflammation, fibrosis, and a series of pathological damage[3-4].The suppressors of cytokine sigmaling-1 (SOCS-1)is a key negative regulatory factor of JAK/STAT signal transduction pathway. In the kidney, the high expression of SOCS-1 can reduce the pathological damage of DN. At the same time, vascular endothelial growth factor (VEGF) is also involved in the pathogenesis of DN, in recent years, the study found that the activation of JAK/STAT signaling pathway can regulate the expression of VEGF [5]. Tangshen Heji is the compound Chinese medicine, it is based on many years clinical experience summed up by professor mentor Yao Yuanzhang. Tangshen Heji have obtained good curative effect in the treatment of patients with diabetic nephropathy.The formula relevancy DN pathogenesis of traditional Chinese medicine, it is based on deficiency of qi and Yin, both moist heat、blood stasis、poison, it has owned the efficacy of nourish qi to generate fluid and promote blood circulation to remove meridian obstruction. In the early stage of the work, we through the experiment proves that part of traditional Chinese medicine (TCM) in Tangshen Heji can be adjusted through JAK/STAT pathway to protect the kidney. But the Tangshen Heji’s overall curative effect is relevant to regulating the expression of SOCS-1, TGF-beta 1,VEGF in kidney tissues? The test’s purpose is to look at the effect of Tangshen Heji both high and low dose in diabetic rat kidney and the influence to the expression of SOCS-1, TGF-beta 1 VEGF in kidney tissues. And angiotensin Ⅱ (Ang Ⅱ) type 1 receptor antagonist Losartan is sanded as positive control.The pursose is to explore Tangshen Heji’s protection for diabetic rats kidney, and its possible mechanism.The research methods1. The establishment of the diabetic rats model:using high fat diet in combination with small dose of streptozotocin (STZ) intraperitoneal injection;the standard of model is non fasting glucose> 16.7 mmol/L.2. Grouping:SD rats were randomly divided into normal control group (N group), diabetic group (DM group), Tangshen Heji low dose group (TS1 group), Tangshen Heji high dose group (TS2group), Losartan group (L group).3. The dosing methods:normal group, diabetes group rats are given Sodium chloride by 15 ml/Kg/d dose, Tangshen Heji high and low dose group and Losartan group rats are given corresponding solution by 15 ml/Kg/d dose. Drug intervention will keep on a total of 6 weeks.4. The curative effect evaluation method:determinate of each rat BM, BG,24 h urine protein quantitative; After anesthesia,we can cull blood from abdominal aorta to detect the serum creatinine (Scr), urea nitrogen (BUN), serum albumin (ALB); Get kidney,then renal tissue paraffin blade to dyeing hematoxylin eosin (HE), renal tissue pathological changes were observed under light microscope; Using immunohisto-chemical and Western blot method to detect the expression of SOCS-1, TGF-beta 1, VEGF protein in diabetic rat kidney.Using Image analysis software (Image-Pro Plus) semi-quantitative analysis, then appling SPSS 17.0 statistical analysis software for experimental data statistics processing progressively.We seem P < 0.05 as statistically significant, P< 0.01 as significant statistical significance.Results1. The 24 h urine protein quantitative, serum creatinine, blood urea nitrogen of diabetes group were higher than normal group (P< 0.01), serum albumin lower than normal group (P< 0.01); Compared with diabetes group, the 24 h urine protein quantitative of Tangshen Heji high and low dose group, Losartan group were significantly reduced (P< 0.01), serum albumin were significantly increased (P< 0.01), the serum creatinine, blood urea nitrogen of Tangshen Heji high and low dose group were declined comparing with diabetic group2. The pathological changes:It is microscopical cvisible that normal group rat glomerular structure is clear, morphological rules. In diabetes group,the rats glomerular mesangial matrix appear a moderately and severe hyperplasia and a diffuse change, thickening of basement membrane, balloon part adhesion.Renal tubular epithelial cells happen the vacuoles degeneration, interstitial inflammatory cells occur the infiltration and fibrosis. Glomerular lesions in the treatment group were compared with diabetes group, and the effect of Tangshen Heji high dose group and Losartan group is obviously.3. In diabetes group, SOCS-1,TGF-beta 1, VEGF protein expression were significantly increased than the normal group (P< 0.01);Compared with diabetes group, SOCS-1 expression of Tangshen Heji high dose group, Losartan group is further increase (P< 0.01); TGF-beta 1, VEGF expression of Tangshen Heji high and low dose group, Losartan group are decreased (P< 0.01); Compared with Tangshen Heji low dose group, in the rat kidney tissue, TGF-beta 1, VEGF expression of Tangshen Heji high dose group, Losartan group are further reduce (P< 0.05).ConclusionTangshen Heji can effectively reduce diabetic rats urinary protein, serum creatinine and urea nitrogen, increase serum albumin, can reduce diabetic rats renal tissue pathological changes, this protective effect of kidney may be related to raise the expression of local renal tissue SOCS-1, negative regulation of JAK/STAT signaling pathway, inhibition of TGF-beta 1, VEGF expression.