加味黄风汤对糖尿病肾病大鼠肾组织SIRT1及PGC-1α表达的影响
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摘要
目的:观察加味黄风汤对糖尿病肾病(DN)大鼠肾脏组织沉默信息调节因子1(SIRT1)及过氧化物酶体增殖物激活受体γ共激活因子-1α(PGC-1α)表达的影响。方法:雄性SD大鼠经腹腔注射链脲佐菌素(STZ)造模,随机分为正常组、模型组、白藜芦醇组、加味黄风汤低、高剂量组(简称低、高剂量组),各10只,8周后检测24h尿微量白蛋白,光镜及电镜下观察肾脏病理改变,Western Blot方法检测肾组织SIRT1及PGC-1α蛋白的表达水平,RT-PCR方法检测肾组织SIRT1及PGC-1αmRNA的表达水平。结果:与正常组比较,模型组大鼠可见微量白蛋白尿增加,肾脏病理可见肾小球肥大,系膜基质增生,基底膜增厚及足细胞足突融合,肾组织中SIRT1及PGC-1α蛋白及mRNA表达水平显著下降(P<0.05)。8周后,与模型组比较,高、低剂量组DN大鼠24h尿微量白蛋白水平显著下降(P<0.05);光镜下肾小球肥大、系膜基质增生减轻;电镜下足细胞足突融合、基底膜增厚情况有所改善;肾组织中SIRT1及PGC-1α蛋白及mRNA表达水平均显著上升(P<0.05)。结论:加味黄风汤可降低DN大鼠尿微量白蛋白,改善肾脏病理损伤,其机制可能与上调肾组织的SIRT1及PGC-1α的表达水平,改善足细胞能量代谢相关。
Objective: To observe the effects of Jiawei Huangfeng Decoction on the expression of SIRT1 and PGC-1αin renal tissue of rats with diabetic nephropathy(DN). Methods: The male SD rats were established by injecting intraperitoneally streptozotocin(STZ), then rats were randomly divided into control group, model group, resveratrol group, Jiawei Huangfeng Decoction low dose group and high dose group, 10 rats in each one. After 8 weeks, the 24-hour urine micro-albumin was detected, and the renal pathological changes were oberved by light microscope and electron microscope. The protein expression of SIRT1 and PGC-1α in renal tissue was detected by Western Blot method. And the mRNA expression of SIRT1 and PGC-1α in renal tissue was detected by RT-PCR. Results: Compared with control group, micro-albumin urine was increased in the model group. Glomerular hypertrophy, mesangial matrix hyperplasia, basement membrane thickening and podocyte foot process fusion were observed in renal pathology of model group. The protein and mRNA expression of SIRT1 and PGC-1 decreased significantly in renal tissue(P<0.05). After treatment for 8 weeks, compared with model group, 24-hour urine micro-albumin of rats with DN was significantly decreased in Jiawei Huangfeng Decoction low dose group and high dose group(P<0.05). Glomerular hypertrophy and mesangial matrix hyperplasia were alleviated under light microscope. Electron microscope observed basement membrane thickening and podocyte foot process fusion had been improved. Compared with model group, the protrin and mRNA expression of SIRT1 and PGC-1α in renal tissue were significantly increased(P<0.05). Conclusion: Jiawei Huangfeng Decoction can reduce urine micro-albumin of rats with DN, improve the pathological damage of DN, and its mechanism may be related to up-regulating the expression of SIRT1 and PGC-1α in renal tissues and improving the energy metabolism of podocytes.
Objective: To observe the effects of Jiawei Huangfeng Decoction on the expression of SIRT1 and PGC-1αin renal tissue of rats with diabetic nephropathy(DN). Methods: The male SD rats were established by injecting intraperitoneally streptozotocin(STZ), then rats were randomly divided into control group, model group, resveratrol group, Jiawei Huangfeng Decoction low dose group and high dose group, 10 rats in each one. After 8 weeks, the 24-hour urine micro-albumin was detected, and the renal pathological changes were oberved by light microscope and electron microscope. The protein expression of SIRT1 and PGC-1α in renal tissue was detected by Western Blot method. And the mRNA expression of SIRT1 and PGC-1α in renal tissue was detected by RT-PCR. Results: Compared with control group, micro-albumin urine was increased in the model group. Glomerular hypertrophy, mesangial matrix hyperplasia, basement membrane thickening and podocyte foot process fusion were observed in renal pathology of model group. The protein and mRNA expression of SIRT1 and PGC-1 decreased significantly in renal tissue(P<0.05). After treatment for 8 weeks, compared with model group, 24-hour urine micro-albumin of rats with DN was significantly decreased in Jiawei Huangfeng Decoction low dose group and high dose group(P<0.05). Glomerular hypertrophy and mesangial matrix hyperplasia were alleviated under light microscope. Electron microscope observed basement membrane thickening and podocyte foot process fusion had been improved. Compared with model group, the protrin and mRNA expression of SIRT1 and PGC-1α in renal tissue were significantly increased(P<0.05). Conclusion: Jiawei Huangfeng Decoction can reduce urine micro-albumin of rats with DN, improve the pathological damage of DN, and its mechanism may be related to up-regulating the expression of SIRT1 and PGC-1α in renal tissues and improving the energy metabolism of podocytes.
引文
[1]米乐,张史昭,张培培.加味黄风汤为主治疗风湿内扰型糖尿病肾病30例临床观察.浙江中医杂志,2016,51(8):579
[2]张培培,陈红波,鲁科达,等.加味黄风汤对IgA肾病大鼠肾组织α-SMA及TGF-β表达的影响.中华中医药学刊,2017,35(3):673-676
[3]方莉,张培培,马红珍.加味黄风汤对糖尿病肾病足细胞损伤保护机制研究进展.辽宁中医药大学学报,2016,18(12):121-123
[4]Xu Y,Nie L,Yin Y G,et al.Resveratrol protects against hyperglycemiainduced oxidative damage to mitochondria by activating SIRT1in rat mesangial cells.Toxicology&Applied Pharmacology,2012,259(3):395-401
[5]Sharma S,Anjaneyulu M,Kulkarni S K,et al.Resveratrol,apolyphenolic phytoalexin phytoalexin,attenuates diabetic nephropathy in rats.Pharmacology,2006,76(2):69-75
[6]Zhu G Y,Shi B Z,Li Y.FoxM1 regulates Sirt1 expression in glioma cells.European Review for Medical&Pharmacological Sciences,2014,18(2):205-211
[7]Mantel C,Broxmeyer H E.Sirtuin 1,stem cells,aging,and stem cell aging.Current Opinion in Hematology,2008,15(4):326-331
[8]童楠,张宁.中药益肾颗粒通过PI3K/Akt/mTOR和LKB1/AMPK/Sirt1信号通路对糖尿病肾病大鼠的干预作用研究.中华中医药杂志,2018,33(5):1853-1857
[9]Valle I,Alvarez-Barrientos A,Arza E.PGC-1alpha regulates the mitochondrial antioxidant defense system in vascular endothelial cells.Cardiovasc Res,2005,66(3):562-573
[10]Summermatter S,Shui G,Maag D,et al.PGC-1αimproves glucose homeostasis in skeletal muscle in an activity-dependent manner.Diabetes,2013,62(1):85-95
[11]Mitra R,Nogee D P,Zechner J F.The transcriptional coactivators,pgc-1alpha and beta,cooperate to maintain cardiac mitochondrial function during the early stages of insulin resistance.J Mol Cell Cardiol,2012,52(3):701-710
[12]Lakshmanan A P,Watanabe K,Thandavarayan R A.Telmisartan attenuates oxidative stress and renal fibrosis in streptozotocin induced diabetic mice with the alteration of angiotensin-(1-7)mas receptor expression associated with its PPAR-gamma agonist action.Free Radic Res,2011,45(5):575-584
[13]Funk J A,Odejinmi S,Schnellmann R G.SRT1720 induces mitochondrial biogenesis and rescues mitochondrial function after oxidant injury in renal proximal tubule cells.J Pharmacol Exp Ther,2010,333(2):593-601
[2]张培培,陈红波,鲁科达,等.加味黄风汤对IgA肾病大鼠肾组织α-SMA及TGF-β表达的影响.中华中医药学刊,2017,35(3):673-676
[3]方莉,张培培,马红珍.加味黄风汤对糖尿病肾病足细胞损伤保护机制研究进展.辽宁中医药大学学报,2016,18(12):121-123
[4]Xu Y,Nie L,Yin Y G,et al.Resveratrol protects against hyperglycemiainduced oxidative damage to mitochondria by activating SIRT1in rat mesangial cells.Toxicology&Applied Pharmacology,2012,259(3):395-401
[5]Sharma S,Anjaneyulu M,Kulkarni S K,et al.Resveratrol,apolyphenolic phytoalexin phytoalexin,attenuates diabetic nephropathy in rats.Pharmacology,2006,76(2):69-75
[6]Zhu G Y,Shi B Z,Li Y.FoxM1 regulates Sirt1 expression in glioma cells.European Review for Medical&Pharmacological Sciences,2014,18(2):205-211
[7]Mantel C,Broxmeyer H E.Sirtuin 1,stem cells,aging,and stem cell aging.Current Opinion in Hematology,2008,15(4):326-331
[8]童楠,张宁.中药益肾颗粒通过PI3K/Akt/mTOR和LKB1/AMPK/Sirt1信号通路对糖尿病肾病大鼠的干预作用研究.中华中医药杂志,2018,33(5):1853-1857
[9]Valle I,Alvarez-Barrientos A,Arza E.PGC-1alpha regulates the mitochondrial antioxidant defense system in vascular endothelial cells.Cardiovasc Res,2005,66(3):562-573
[10]Summermatter S,Shui G,Maag D,et al.PGC-1αimproves glucose homeostasis in skeletal muscle in an activity-dependent manner.Diabetes,2013,62(1):85-95
[11]Mitra R,Nogee D P,Zechner J F.The transcriptional coactivators,pgc-1alpha and beta,cooperate to maintain cardiac mitochondrial function during the early stages of insulin resistance.J Mol Cell Cardiol,2012,52(3):701-710
[12]Lakshmanan A P,Watanabe K,Thandavarayan R A.Telmisartan attenuates oxidative stress and renal fibrosis in streptozotocin induced diabetic mice with the alteration of angiotensin-(1-7)mas receptor expression associated with its PPAR-gamma agonist action.Free Radic Res,2011,45(5):575-584
[13]Funk J A,Odejinmi S,Schnellmann R G.SRT1720 induces mitochondrial biogenesis and rescues mitochondrial function after oxidant injury in renal proximal tubule cells.J Pharmacol Exp Ther,2010,333(2):593-601