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Objective: Diabetes (diabetes mellitus, DM) can cause the retina, kidneys, nerves and other organs of complications, including diabetic nephropathy (diabetic nephropathy, DN) is the most serious chronic complications of diabetes, the incidence rate is increasing every year. DN early primary pathology is characterized by glomerular basement membrane thickening, mesangial cell proliferation, 'mesangial expansion induced renal hypertrophy, clinical manifestations of high filtration and microalbuminuria. With the progression of glomerular mesangial matrix increase, promote glomerulosclerosis and tubulointerstitial fibrosis, decreased kidney function, eventually leading to end-stage renal failure. Currently, in vivo experiments show that extracellular signal-regulated protein (extracellular signal regulated kinase, ERK) signaling pathway in diabetic kidney disease occurrence and development plays an important role. ERK can be a variety of growth factors and cytokines activation mediates cell growth, proliferation, differentiation. Angiotensin Ⅱ (Ang Ⅱ) can stimulate vascular smooth muscle cells and mesangial cells, renal tubular epithelial cell hypertrophy. Vascular endothelial growth factor (vascular endothelial growth factor, VEGF) is a promotion of endothelial cell proliferation and migration, increased vascular permeability cytokines. CTGF (connective tissue growth factor, CTGF) is to promote the fibrotic cytokines. Advanced glycation end products (advanced glycation end products, AGEs) and receptor of advanced glycation end products (advanced glycation end products receptor, RAGE) can be combined to facilitate the release of various cytokines, causing endothelial damage, extracellular stromal hyperplasia and other pathological changes. Ang Ⅱ, VEGF, CTGF, AGEs and ERK signal transduction pathways are closely related. In vitro experiments have confirmed, Ang Ⅱ, VEGF, AGEs can activate the ERK pathway kidney cells, promote the expression of CTGF. Tongluo prescription main ingredient is ginseng, leeches, scorpion, woodlouse, centipede, TPG and other drugs. The experimental discovery through TLR has improved glomerular basement membrane and high-sugar induced oxidative damage and increase the role of antioxidant capacity, can reduce renal tissue Ang Ⅱ, AGEs are generated, but can influence through TLR intracellular signaling transduction pathway is unclear. This study was to study Tongluo Recipe on STZ-induced diabetic rat kidney Ang Ⅱ, VEGF, CTGF, RAGE, ERK1 / 2, phosphorylated ERK1 / 2 (p-ERK1 / 2) expression, as well as to protect the kidneys through TLR possible mechanisms. Methods: SPF level adult male SD rats weighing 200g ± 20g, randomization: modeling group was given a single intraperitoneal injection of STZ60mg/kg, 72 hours after tail vein blood test random blood glucose to glucose ≥ 16.7mmol / L identified as diabetic model, the normal group (A) 10. Diabetic rats were randomly divided into a mold five groups: non-treated diabetic group (B) 10, a low-dose through TLR group (C, 0.5g / (kg · d)) 10 only, and the dose through TLR group (D group, 1.0g / (kg · d)) 10 only, and high-dose through TLR group (E, 2.0g / (kg · d)) 10 only, and losartan group (F, 30mg / ( kg · d)) 10 只. Diabetic rats after a successful modeling adaptive feeding one week later, were given different treatment according to the grouping factor, A group and B group received an equal volume of saline, regular daily gavage, continuous administration were sacrificed after 12 weeks, said amount of weight, blood glucose testing (GLU), lipids (TG, CHOL, HDLC, LDLC), serum creatinine (BCREA), blood urea nitrogen (BUN), leaving 24-hour urine test urine creatinine (CREA), urinary protein (UPRO24) microalbuminuria (UMA), take the left kidney weight (KW), calculate the kidney weight / body weight ratio (KW / BW), transmission electron microscopy as well as in HE, PAS light microscope after staining with pathological changes, Elisa method serum and kidney tissue Ang Ⅱ content, immunohistochemical detection of p-ERK1 / 2 protein expression, Western blot assay VEGF, RAGE, CTGF, ERK1 / 2 and p-ERK1 / 2 protein expression, SYBR Green real -time PCR method for detection of renal tissue CTGF mRNA, RAGE mRNA expression. Results: 1, after the model of the weight of the rats compared with normal rats were significantly reduced (P lt; 0.05), C, D, E, F treatment group increased body weight compared with non-treated group, in which a large group D rat body weight significantly increased compared with group B (P lt; 0.05). Modeling diabetes kidney index in each group were significantly higher than normal control (P lt; 0.05), the diabetes treatment group compared to diabetic renal hypertrophy index decreased non-treatment group, in which D, E, F group than in the non-treatment group was significantly decreased (P lt; 0.05). 2 diabetes rats in each group molded GLU, TG was significantly higher than those in the normal group (P lt; 0.05). The diabetic rats as modules HDLC, LDLC compared with the normal group was significantly lower (P lt; 0.05). The treatment of diabetes and diabetic rats non-treatment groups GLU, TG, HDLC, LDLC were not statistically different (P gt; 0.05). CHOL rats in each group was not statistically significant. 3, diabetic rats molded BUN, Ccr, UPRO24, UMA significantly higher than those in the normal group (P lt; 0.05), diabetes treatment groups BUN, Ccr, UMA non-treated group was significantly lower than (P lt; 0.05) , D, E, F 24-hour urine protein in the treatment group compared with non-treated group was significantly decreased (P lt; 0.05), C group UPRO24 reduced compared with non-treated group, but no significant difference. 4, ordinary light microscope no pathological changes in group A, B group the glomerular basement membrane thickening, mesangial cell proliferation, cloudy swelling of renal tubular degeneration, mild renal interstitial fibrosis. Diabetes treatment groups than in group B have different degrees of improvement. Under TEM Group B basement membrane thickening, foot process fusion, fracture glomerular endothelial cells, renal tubular epithelial cell degeneration and necrosis, C, D, E, F group than in group B lesions light. 5, B, C, E, F Ang Ⅱ levels in serum was significantly higher than group A (P lt; 0.05), D group than in group A increased, but not statistically significant. Each treatment group compared with non-treated serum Ang Ⅱ were significantly lower (P lt; 0.05). C, E serum Ang Ⅱ levels higher than the F group were significantly different (P lt; 0.05), D serum levels and Ang Ⅱ A, F group had no significant difference. The diabetic rat kidney tissue as modules Ang Ⅱ levels were significantly higher than normal (P lt; 0.05), diabetes, kidney each treatment group content of Ang Ⅱ decreased significantly compared with untreated diabetes (P lt; 0.05), D, E kidney Ang Ⅱ group F group content was not statistically significant (P gt; 0.05). 6, B, C, D, E, F group, VEGF, RAGE, CTGF, p-ERKl / 2 protein was significantly increased compared with group A (P lt; 0.05), C, D, E, F group, VEGF, RAGE, CTGF, p-ERK1 / 2 protein expression decreased compared with group B (P lt; 0.05), each group Total-ERK1 / 2 protein expression differences between the groups was not statistically significant (P gt; 0.05). 7, D, E, F rat kidney tissue RAGE mRNA levels decreased significantly compared with group B (P lt; 0.05). D, E rat kidney tissue levels of CTGF mRNA compared with group B, F group was significantly lower (P lt; 0.05). Compared with group B, C rat kidney RAGE mRNA, CTGF mRNA levels decreased, but no significant difference (P gt; 0.05). Conclusions: 1, diabetic nephropathy and renal pathological structure damage. 2, can be reduced through TLR pathological changes in diabetic rats, improve kidney function, kidney meridians prescription protection similar to the role and losartan. 2, the kidney meridians prescription protective mechanism may be related to inhibition of Ang Ⅱ, VEGF, RAGE expression, blocking ERK1 / 2 signaling pathway, reducing the expression of CTGF delay related to the process of renal fibrosis.
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