ABSTRACT
Objective To observe the effects of Tongluo reeipe on extracellular signal-regulated protein kinase (ERK1/2) pathway in the kidneys of streptozotocin-induced rat models of type 1 diabetes mellitus, and to explore the renoprotective mechanism of Tongluo reeipe. Methods Adult male Sprague-Dawley rats were randomly divided into 2 groups: control group (A, n=10) and experimental group. Rats in the experimental group were induced by intraperitoneal injection of streptozotocin (60 mg/kg body mass), and were randomly subdivided into group B (diabetic control, n=10), group C (diabetic rats treated with low dose of Tongluo recipe,0.5g/[kg · d], n=10), group D (diabetic rats treated with medium dose of Tongluo recipe, 1.0g/[kg · d], n=10), group E (diabetic rats treated with high dose of Tongluo recipe, 2.0g/[kg · d], n=10) and group F (diabetic rats treated with losartan, 30 mg/[kg · d],n=10). At the end of the 12th week rats were sacrificed, and the serum glucose, triacylglycerol(TG), total cholesterol(Chol), serum creatinine(Scr), urea nitrogen(BUN) in blood, 24-hour urine protein(Upro24), microalbuminuria (MAU), and creatinine in urine (UCR) were examined. The kidney mass/body mass (KM/BM) and creatinine clearance rate (Ccr) were calculated. The left kídneys were collected, weighed and subjected to H-E staining and PAS staining to observe the pathological changes; serum and renal tissue angiotensin II (Ang II) levels were determined by ELISA; the expressíon of phosphorylated ERK1/2 (p-ERK1/2) was detected by immunohistochemical staining and vascular endothelial growth factor (VEGF) protein expression was determined by Western blotting analysis. Results Compared with group B, group D had a slighter mesangial proliferation under light microscope. Besides, rats in group D had significantly decreased Ccr, BUN, MAU, and Upro24 in the serum, and decreased Ang II in the serum and renal tissue, and decreased KM/BM, VEGF and p-ERK1/2 levels in renal tissue compared with rats in group B(P<0.05). The medium dose of Tongluo recipe and losartan showed similar effects in improving the renal function and inhibiting Ang II, VEGF, and p-ERK1/2 expression of diabetic rats. Conclusion The renoprotective effect of Tongluo recipe may be related to inhibition of ERK1/2 cascade pathway by decreasing the expression of Ang II and VEGF.
ABSTRACT
Objective To observe the effects of Tongluo recipe on extracellular signal-regulated protein kinase (ERK1/2) pathway in the kidneys of streptozotocin-induced rat models of type 1 diabetes mellitus, andto explore the renoprotective mechanism of Tongluo recipe. Methods Adult male Sprague-Dawley rats were randomly divided into 2 groups: control group (A, n = 10) and experimental group. Rats in the experimental group were induced by intraperitoneal injection of streptozotocin (60 mg/kg body mass), and were randomly subdivided into group B (diabetic control, n = 10), group C (diabetic rats treated with low dose of Tongluo recipe, 0. 5 g/[kg • d], n = 10), group D (diabetic rats treated with medium dose of Tongluo recipe, 1. 0 g/[kg • d], n = 10), group E (diabetic rats treated with high dose of Tongluo recipe,2. 0 g/[kg • d], n = 10) and group F (diabetic rats treated with losartan, 30 mg/[kg • d],n=10). At the end of the 12th week rats were sacrificed, and the serum glucose, triacylglycerol(TG), total cholesterol(Choi), serum creatinine(Scr), urea nitrogen(BUN) in blood, 24-hour urine protein(Upro24), microalbuminuria (MAU), and creatinine in urine(UCR) were examined. The kidney mass/body mass (KM/BM) and creatinine clearance rate (Ccr) were calculated. The left kidneyswere collected, weighed and subjected to H-E staining and PAS staining to observe the pathological changes; serum and renal tissue angiotensin II (Ang II) levels were determined by ELISA; the expression of phosphorylated ERK1/2(p-ERK1/2) was detectedby immunohistochemical staining and vascular endothelial growth factor (VEGF) protein expression was determined by Western blotting analysis. Results Compared with group B, group D had a slighter mesangial proliferation under light microscope. Besides, rats in group D had significantly decreased Ccr, BUN, MAU, and Upro24 in the serum, and decreased Ang II in the serum and renal tissue, and decreased KM/BM, VEGF and p-ERK1/2 levels in renal tissue compared with rats in group B(P<0. 05). The medium dose of Tongluo recipe and losartan showed similar effects in improving the renal function and inhibiting Ang II, VEGF, and p-ERK1/2 expression of diabetic rats. Conclusion The renoprotective effect of Tongluo recipe may be related to inhibition of ERK1/2 cascade pathway by decreasing the expression of Ang II and VEGF.
ABSTRACT
Objective: To observe the effects of Tongluo recipe on the renal expression of connective tissue growth factor (CTGF), advanced glycation end products receptor (RAGE) and phosphorylated extracellular signal regulated kinase (p-ERK1/2) in streptozotocin-induced rat model of type 1 diabetes mellitus, and to explore the mechanism of its renoprotective effect. Methods: Adult male Sprague-Dawley rats, weighing 180-220 g, were randomly divided into 2 groups: control group (A, n=10) and model group (n=50). Rat diabetic model was induced by intraperitoneal injection of streptozotocin (60 mg/kg body weight), and the model rats were randomly subdivided into B group (diabetic control, n=10), C group (diabetic rats treated with low dose of Tongluo recipe, 0.5 g·kg-1·d-1, n=10), D group (diabetic rats treated with medium dose of Tongluo recipe, 1.0 g·kg -1·d-1, n=10), E group (diabetic rats treated with high dose of Tongluo recipe, 2.0 g·kg-1·d-1, n=10), and F group (diabetic rats treated with losartan, 30 g·kg -1·d-1, n=10). At the end of the 12th week the rats were sacrificed and were weighed; the urea nitrogen (BUN) in blood, serum creatinine (Scr), 24 h albumin excretion rate (UPRO24), microalbuminuria (MAU) and creatinine in urine (Ucr) were analyzed by bio-chemical methods. The left kidneys were weighed and the right kidneys were subjected to real-time PCR analysis for the expression of CTGF and RAGE mRNA. Western blotting analysis was used to examine the expression of CTGF, RAGE, p-ERK1/2, and total-ERK1/2 protein expression. Results: The creatinine clearance rate, urea nitrogen, 24 h albumin excretion rate, and microalbuminuria in group D and E were significantly lower than those of group B. And the renal functions of rats in group D and E were improved and the expression of RAGE and CTGF mRNA and protein and p-ERK1/2 protein expression was significantly decreased (P<0.05). Conclusion: Tongluo recipe may exert its renoprotective effect through inhibiting RAGE-ERK1/2-CTGF signal pathway.