丹酚酸B对糖尿病大鼠肾纤维化的改善作用及其机制
|
摘要
目的观察丹酚酸B对糖尿病大鼠肾纤维化的影响,并探讨其可能机制。方法采用高糖高脂饮食,合并腹腔注射链脲佐菌素(STZ)的方法建立糖尿病大鼠模型。将糖尿病大鼠随机分为模型组、丹酚酸B低、高剂量组(80、160 mg·kg~(-1))。丹酚酸B各组每天灌胃给予相应剂量药物,正常对照组和模型组灌胃相应容量生理盐水,连续6周。实验结束时检测大鼠空腹血糖(FBG)、血清肌酐(SCr)、尿素氮(BUN)、尿微量白蛋白(UAlb),以及血清丙二醛(MDA)、总抗氧化能力(TAC)和超氧化物歧化酶(SOD)水平;Masson染色法观察肾组织胶原纤维水平;ELISA法检测肾组织Ⅰ型和Ⅲ型胶原含量;Western blot法检测肾组织转化生长因子β1(TGF-β1)、p-Smad2和Smad7蛋白水平。结果丹酚酸B可明显减少糖尿病大鼠肾组织胶原沉积,降低FBG、BUN、UAlb、SCr、MDA及TGF-β1、p-Smad2蛋白水平,使血清TAC、SOD活性和肾组织Smad7蛋白水平明显升高(P<0.01或P<0. 05)。结论丹酚酸B可明显减轻糖尿病大鼠肾纤维化,改善肾功能,其机制与改善氧化应激状态、调控TGF-β1/Smad信号通路有关。
Aim To investigate the ameliorative effect of salvianolic acid B(Sal B) on renal fibrosis in diabetic rats and the possible mechanisms implicated. Methods Diabetes in SD rats was induced by high sugar high fat diet combined with intraperitoneal injection of streptozotocin(STZ, 40 mg·kg~(-1)). Diabetic rats were allocated randomly to model group, high dose of Sal B group(160 mg·kg~(-1)) and low dose of Sal B group(80 mg·kg~(-1)). Rats in Sal B groups were administered intragastrically with corresponding dose of Sal B for six weeks, while those in the control and model group were given distilled water of the same volume. At the end of experiment, the levels of fasting blood glucose(FBG), serum creatinine(SCr), blood urea nitrogen(BUN), urine microalbumin(UAlb), serum malondialdehyde(MDA), total antioxidant capacity(TAC) and superoxide dismutase(SOD) were determined with commercially available kits. Renal fibrosis was evaluated by Masson staining and renal contents of type I and type III collagen determined by ELISA. The protein levels of TGF-β1, p-Smad2 and Smad7 were evaluated by Western blot analysis. Results In model group, the levels of FBG, SCr, BUN, UAlb and MDA significantly increased as compared with those in control group, while TAC and SOD activities decreased markedly(P<0.01). Rats in model group exhibited evident renal fibrosis, accompanied by significantly increased levels of TGF-β1 and p-Smad2 protein and decreased level of Smad7 protein in renal tissues(P<0.01). Supplementation with Sal B significantly ameliorated renal fibrosis in diabetic rats, with TGF-β1 and p-Smad2 protein down-regulated and Smad7 protein up-regulated evidently(P<0.05 or P<0.01). The above mentioned changes in biochemical parameters were also reversed markedly in Sal B treated groups(P<0.05 or P<0.01). Conclusions Sal B is capable of ameliorating renal fibrosis in diabetic rats, which might be related to its antioxidant property and regulation of TGF-β1/Smad signaling pathway.
Aim To investigate the ameliorative effect of salvianolic acid B(Sal B) on renal fibrosis in diabetic rats and the possible mechanisms implicated. Methods Diabetes in SD rats was induced by high sugar high fat diet combined with intraperitoneal injection of streptozotocin(STZ, 40 mg·kg~(-1)). Diabetic rats were allocated randomly to model group, high dose of Sal B group(160 mg·kg~(-1)) and low dose of Sal B group(80 mg·kg~(-1)). Rats in Sal B groups were administered intragastrically with corresponding dose of Sal B for six weeks, while those in the control and model group were given distilled water of the same volume. At the end of experiment, the levels of fasting blood glucose(FBG), serum creatinine(SCr), blood urea nitrogen(BUN), urine microalbumin(UAlb), serum malondialdehyde(MDA), total antioxidant capacity(TAC) and superoxide dismutase(SOD) were determined with commercially available kits. Renal fibrosis was evaluated by Masson staining and renal contents of type I and type III collagen determined by ELISA. The protein levels of TGF-β1, p-Smad2 and Smad7 were evaluated by Western blot analysis. Results In model group, the levels of FBG, SCr, BUN, UAlb and MDA significantly increased as compared with those in control group, while TAC and SOD activities decreased markedly(P<0.01). Rats in model group exhibited evident renal fibrosis, accompanied by significantly increased levels of TGF-β1 and p-Smad2 protein and decreased level of Smad7 protein in renal tissues(P<0.01). Supplementation with Sal B significantly ameliorated renal fibrosis in diabetic rats, with TGF-β1 and p-Smad2 protein down-regulated and Smad7 protein up-regulated evidently(P<0.05 or P<0.01). The above mentioned changes in biochemical parameters were also reversed markedly in Sal B treated groups(P<0.05 or P<0.01). Conclusions Sal B is capable of ameliorating renal fibrosis in diabetic rats, which might be related to its antioxidant property and regulation of TGF-β1/Smad signaling pathway.
引文
[1] Magee C, Grieve D J, Watson C J, et al. Diabetic nephropathy: a tangled web to unweave [J]. Cardiovasc Drugs Ther, 2017, 31(5-6):579-92.
[2] Sun J, Wang Y W, Cui W P, et al. Role of epigenetic histone modifications in diabetic kidney disease involving renal fibrosis [J]. J Diabetes Res, 2017, 2017:7242384.
[3] Kandhare A D, Mukherjee A, Bodhankar S L. Antioxidant for treatment of diabetic nephropathy: a systematic review and meta-analysis [J]. Chem Biol Interact, 2017, 278:212-21.
[4] Wang D D, Zhang G Y, Chen X, et al. Sitagliptin ameliorates diabetic nephropathy by blocking TGF-β1/Smad signaling pathway [J]. Int J Mol Med, 2018, 41(5):2784-92.
[5] 郑书国,朱元美,陶善珺,等. 丹酚酸 B 对间歇性高糖诱导的JNK活化和INS-1细胞凋亡的影响[J]. 中国药理学通报, 2017, 33(1):68-73.
[5] Zheng S G, Zhu Y M, Tao S J, et al. Effect of salvianolic acid B on intermittent high glucose induced JNK activation and INS-1 cell apoptosis [J]. Chin Pharmacol Bull, 2017, 33(1):68-73.
[6] 张昌志,石明隽,王圆圆,等. 丹酚酸 B 对高糖培养肾小管上皮细胞转分化的影响及意义[J]. 贵阳医学院学报, 2015, 40(4):337-45.
[6] Zhang C Z, Shi M J, Wang Y Y, et al. The effect of salvianolic acid B on high- glucose induced renal tubule epithelial cell epithelial-mesenchymal transition[J]. J Guiyang Med Coll, 2015, 40(4):337-45.
[7] Weng H B, Han W K, Xiong Y W, et al. Taxus chinensis ameliorates diabetic nephropathy through down-regulating TGF-β1/Smad pathway [J]. Chin J Nat Med, 2018, 16(2):90-6.
[8] Lan H Y. Transforming growth factor-β/Smad signalling in diabetic nephropathy [J]. Clin Exp Pharmacol Physiol, 2012, 39(8):731-8.
[9] Walton K L, Johnson K E, Harrison C A. Targeting TGF-β mediated SMAD signaling for the prevention of fibrosis [J]. Front Pharmacol, 2017, 8:461.
[10] 张敏,曹述任. 丹酚酸B对人肺成纤维细胞TGF-β1/Smad信号通路相关蛋白表达的影响及其机制[J]. 吉林大学学报(医学版), 2014, 40(4):705-9.
[10] Zhang M, Cao S R. Influence of salvianolic acid B on expression of TGF-β1/Smad signaling pathway related proteins in human lung fibroblasts and its mechanism [J]. J Jilin Univ (Med Ed), 2014, 40(4):705-9.
[11] 刘煜敏,张悦,陆海英,等. 丹酚酸B对活化的系膜细胞TGF-β1受体和Smad2表达的影响[J]. 中国病理生理杂志, 2010, 26(9):1734-7.
[11] Liu Y M, Zhang Y, Lu H Y, et al. Effect of salvianolic acid B on expression of TGF-β1 receptors and Smad2 in activated mesangial cells [J]. Chin J Pathophysiol , 2010, 26(9):1734-7.
[12] 郭福团,许雄伟,潘建峰,等. 桑枝多糖对糖尿病肾病小鼠肾皮质氧化应激作用的影响[J]. 中国药理学通报, 2016, 32(8):1148-52.
[12] Guo F T, Xu X W, Pan J F, et al. Protective effects of NDP on LPS induced neuron injuries in rat mixed cultures[J]. Chin Pharmacol Bull, 2016, 32(8):1148-52.
[13] Flemming N B, Gallo L A, Forbes J M. Mitochondrial dysfunction and signaling in diabetic kidney disease: oxidative stress and beyond [J]. Semin Nephrol, 2018, 38(2):101-10.
[14] 王超,张会欣,邢邯英,等. 通心络胶囊抑制p38 MAPK 磷酸化抑制糖尿病周围神经病变小鼠氧化应激[J]. 中国药理学通报, 2015, 31(5):726-30.
[14] Wang C, Zhang H X, Xin H Y, et al. Tongxinluo capsule inhibits oxidative stress in diabetic peripheral neuropathy mice by inhibiting the activity of p-p38 MAPK [J]. Chin Pharmacol Bull, 2015, 31(5):726-30.
[15] Al-Waili N, Al-Waili H, Al-Waili T, et al. Natural antioxidants in the treatment and prevention of diabetic nephropathy; a potential approach that warrants clinical trials[J]. Redox Rep, 2017, 22(3):99-118.
[2] Sun J, Wang Y W, Cui W P, et al. Role of epigenetic histone modifications in diabetic kidney disease involving renal fibrosis [J]. J Diabetes Res, 2017, 2017:7242384.
[3] Kandhare A D, Mukherjee A, Bodhankar S L. Antioxidant for treatment of diabetic nephropathy: a systematic review and meta-analysis [J]. Chem Biol Interact, 2017, 278:212-21.
[4] Wang D D, Zhang G Y, Chen X, et al. Sitagliptin ameliorates diabetic nephropathy by blocking TGF-β1/Smad signaling pathway [J]. Int J Mol Med, 2018, 41(5):2784-92.
[5] 郑书国,朱元美,陶善珺,等. 丹酚酸 B 对间歇性高糖诱导的JNK活化和INS-1细胞凋亡的影响[J]. 中国药理学通报, 2017, 33(1):68-73.
[5] Zheng S G, Zhu Y M, Tao S J, et al. Effect of salvianolic acid B on intermittent high glucose induced JNK activation and INS-1 cell apoptosis [J]. Chin Pharmacol Bull, 2017, 33(1):68-73.
[6] 张昌志,石明隽,王圆圆,等. 丹酚酸 B 对高糖培养肾小管上皮细胞转分化的影响及意义[J]. 贵阳医学院学报, 2015, 40(4):337-45.
[6] Zhang C Z, Shi M J, Wang Y Y, et al. The effect of salvianolic acid B on high- glucose induced renal tubule epithelial cell epithelial-mesenchymal transition[J]. J Guiyang Med Coll, 2015, 40(4):337-45.
[7] Weng H B, Han W K, Xiong Y W, et al. Taxus chinensis ameliorates diabetic nephropathy through down-regulating TGF-β1/Smad pathway [J]. Chin J Nat Med, 2018, 16(2):90-6.
[8] Lan H Y. Transforming growth factor-β/Smad signalling in diabetic nephropathy [J]. Clin Exp Pharmacol Physiol, 2012, 39(8):731-8.
[9] Walton K L, Johnson K E, Harrison C A. Targeting TGF-β mediated SMAD signaling for the prevention of fibrosis [J]. Front Pharmacol, 2017, 8:461.
[10] 张敏,曹述任. 丹酚酸B对人肺成纤维细胞TGF-β1/Smad信号通路相关蛋白表达的影响及其机制[J]. 吉林大学学报(医学版), 2014, 40(4):705-9.
[10] Zhang M, Cao S R. Influence of salvianolic acid B on expression of TGF-β1/Smad signaling pathway related proteins in human lung fibroblasts and its mechanism [J]. J Jilin Univ (Med Ed), 2014, 40(4):705-9.
[11] 刘煜敏,张悦,陆海英,等. 丹酚酸B对活化的系膜细胞TGF-β1受体和Smad2表达的影响[J]. 中国病理生理杂志, 2010, 26(9):1734-7.
[11] Liu Y M, Zhang Y, Lu H Y, et al. Effect of salvianolic acid B on expression of TGF-β1 receptors and Smad2 in activated mesangial cells [J]. Chin J Pathophysiol , 2010, 26(9):1734-7.
[12] 郭福团,许雄伟,潘建峰,等. 桑枝多糖对糖尿病肾病小鼠肾皮质氧化应激作用的影响[J]. 中国药理学通报, 2016, 32(8):1148-52.
[12] Guo F T, Xu X W, Pan J F, et al. Protective effects of NDP on LPS induced neuron injuries in rat mixed cultures[J]. Chin Pharmacol Bull, 2016, 32(8):1148-52.
[13] Flemming N B, Gallo L A, Forbes J M. Mitochondrial dysfunction and signaling in diabetic kidney disease: oxidative stress and beyond [J]. Semin Nephrol, 2018, 38(2):101-10.
[14] 王超,张会欣,邢邯英,等. 通心络胶囊抑制p38 MAPK 磷酸化抑制糖尿病周围神经病变小鼠氧化应激[J]. 中国药理学通报, 2015, 31(5):726-30.
[14] Wang C, Zhang H X, Xin H Y, et al. Tongxinluo capsule inhibits oxidative stress in diabetic peripheral neuropathy mice by inhibiting the activity of p-p38 MAPK [J]. Chin Pharmacol Bull, 2015, 31(5):726-30.
[15] Al-Waili N, Al-Waili H, Al-Waili T, et al. Natural antioxidants in the treatment and prevention of diabetic nephropathy; a potential approach that warrants clinical trials[J]. Redox Rep, 2017, 22(3):99-118.