尿毒清颗粒对糖尿病肾病大鼠肾脏抗炎抗氧化保护作用及对TGF-β_1/p38MAPK信号通路影响的研究
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摘要
目的探讨尿毒清胶囊对糖尿病肾病大鼠的抗炎、抗氧化作用及对糖尿病肾病大鼠肾组织TGF-β_1/p38MAPK信号通路的影响。方法 70只雄性SD大鼠,随机分为10只正常组,60只模型组。模型组大鼠腹腔注射45 mg·kg~(-1)链脲佐菌素(STZ)造模,取造模成功的60只大鼠随机分为12只模型组,12只厄贝沙坦阳性组,12只尿毒清颗粒低剂量组(0.04 mg·kg~(-1)),12只尿毒清颗粒中剂量组(0.08 mg·kg~(-1)),12只尿毒清颗粒高剂量(0.16 mg·kg~(-1)),灌胃给药12周,模型组和空白组给予生理盐水。每周称量体质量,每周检测24 h尿蛋白量。给药治疗12周后,取血检测血尿素氮(BUN)、血清肌酐(SCr)、丙二醛(MDA)、一氧化氮(NO)、甘油三酯(TG)、超氧化物歧化酶(SOD)含量;麻醉处死大鼠,各取8只对肾组织行HE染色,观察肾组织的病理形态;并取肾组织进行RT-PCR和免疫组化观察TGF-β_1/p38MAPK信号转导等转化因子的mRNA及蛋白表达。结果与正常组比较,模型组大鼠尿蛋白及血清BUN、SCr、TG、MDA含量显著增高(P<0.05),NO、SOD含量显著降低(P<0.01),肾组织胞浆中TGF-β_1、p38MAPK、Caspase-3 mRNA及蛋白表达显著升高(P<0.05);通过治疗后,与模型组对比,尿毒清颗粒高剂量组和阳性组肾脏病理损害明显减轻,尿蛋白及血清BUN、SCr、TG、MDA含量显著降低(P<0.05),NO、SOD含量明显增高(P<0.05),肾组织内TGF-β_1、p38MAPK、Caspase-3 m RNA及蛋白表达明显减少(P<0.05)。结论尿毒清颗粒能降低24 h尿蛋白量,降低TG和MDA含量及升高NO、SOD含量,通过调控TGF-β_1/p38MAPK从而治疗糖尿病肾病的肾组织受损部位,改善肾功能和减轻病理损伤,能有效地保护糖尿病肾病组织。
Objective To explore effects of uremic granule on the treatment of diabetic nephropathy rats and TGF-β_1/p38 MAPK signaling pathway. Methods Seventy male SD rats were randomly divided into normal group(10 rats),model group(60 rats). After models successfully established by intraperitoneal injection of 45 mg?kg~(-1)streptozotocin(STZ), the model rats were randomly divided into model group, positive control group(irbesartan, 12 rats),uremic granule low,medium,and high dose groups(0.04,0.08,0.16 mg?kg~(-1),12 rats each,respectively).Rats were gavage fed with correspondent drugs for 12 weeks,the model group and normal(blank) group rats weregiven saline solution. The body weights were weighed every week;and 24 h urine protein contents were measuredevery week. At the end of drug treatment,blood urea nitrogen(BUN),serum creatinine(SCr),malondialdehyde(MDA), nitric oxide(NO), serum creatinine(SCr), triglycerides(TG) contents and activity of superoxidedismutase(SOD) in rats' blood were detected. Rats were anesthesia executed,and kidney tissues HE staining of8 rats from each group was carried out to observed the pathological morphology. Transforming growth factor β1(TGF-β_1), p38 MAPK, Caspase 3mRNAs and proteins expression in kidney tissues were detected by RT-PCR andimmunohistochemistry. Results Compared with normal group,urinary protein and serum BUN,SCr,TG and MDAcontents of model group rats increased significantly(P<0.05),NO content and SOD activity decreased significantly(P<0.01); cytoplasm TGF-β_1, p38 MAPK, Caspase 3 mRNA and protein expression of kidney tissues weresignificantly higher(P<0.05). After treatment,compared with model group,uremic granule high-dose group andpositive group showed significantly reduced renal pathological damage,and the urine protein and serum BUN,SCr,TG, MDA contents decreased significantly(P<0.05); NO content and SOD activity increased obviously(P<0.05),TGF-β_1,p38 MAPK,Caspase 3 mRNAs and proteins expression in renal tissues decreased significantly(P<0.05). Conclusion Uremic granule can lower the 24 h urine protein level, lower TG and MDA contents,increase the NO content and SOD activity,effectively protect the diabetic nephropathy,improve renal function andreduce the pathological damage through regulating the TGF-β_1/p38 MAPK signaling pathway.
Objective To explore effects of uremic granule on the treatment of diabetic nephropathy rats and TGF-β_1/p38 MAPK signaling pathway. Methods Seventy male SD rats were randomly divided into normal group(10 rats),model group(60 rats). After models successfully established by intraperitoneal injection of 45 mg?kg~(-1)streptozotocin(STZ), the model rats were randomly divided into model group, positive control group(irbesartan, 12 rats),uremic granule low,medium,and high dose groups(0.04,0.08,0.16 mg?kg~(-1),12 rats each,respectively).Rats were gavage fed with correspondent drugs for 12 weeks,the model group and normal(blank) group rats weregiven saline solution. The body weights were weighed every week;and 24 h urine protein contents were measuredevery week. At the end of drug treatment,blood urea nitrogen(BUN),serum creatinine(SCr),malondialdehyde(MDA), nitric oxide(NO), serum creatinine(SCr), triglycerides(TG) contents and activity of superoxidedismutase(SOD) in rats' blood were detected. Rats were anesthesia executed,and kidney tissues HE staining of8 rats from each group was carried out to observed the pathological morphology. Transforming growth factor β1(TGF-β_1), p38 MAPK, Caspase 3mRNAs and proteins expression in kidney tissues were detected by RT-PCR andimmunohistochemistry. Results Compared with normal group,urinary protein and serum BUN,SCr,TG and MDAcontents of model group rats increased significantly(P<0.05),NO content and SOD activity decreased significantly(P<0.01); cytoplasm TGF-β_1, p38 MAPK, Caspase 3 mRNA and protein expression of kidney tissues weresignificantly higher(P<0.05). After treatment,compared with model group,uremic granule high-dose group andpositive group showed significantly reduced renal pathological damage,and the urine protein and serum BUN,SCr,TG, MDA contents decreased significantly(P<0.05); NO content and SOD activity increased obviously(P<0.05),TGF-β_1,p38 MAPK,Caspase 3 mRNAs and proteins expression in renal tissues decreased significantly(P<0.05). Conclusion Uremic granule can lower the 24 h urine protein level, lower TG and MDA contents,increase the NO content and SOD activity,effectively protect the diabetic nephropathy,improve renal function andreduce the pathological damage through regulating the TGF-β_1/p38 MAPK signaling pathway.
引文
[1]吴航,孙子林.糖尿病肾病的发病机制和药物干预研究进展[J].药学进展,2016,40(5):337-343.
[2]李艳丽,廖勇敢,李晓雯,等.糖尿病肾病发病的危险因素分析[J].实用预防医学,2017,24(2):133-136.
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[4]路文静,武士锋,杨洪涛.尿毒清颗粒治疗糖尿病肾病相关机理研究进展[J].河南中医,2017,37(1):181-184.
[5]张秀峰,张月恒.糖尿病肾病的临床护理方法以及体会[J].中国医药指南,2017,15(4):233-234.
[6]冼启经,曾莉.尿毒清胶囊对梗阻性肾病大鼠肾间质纤维化的影响[J].新中医,2011,43(3):128-130.
[7]乌格敦其其格,赵宗江,蒋玉凤,等.糖肾平胶囊对STZ诱导糖尿病肾病大鼠肾脏保护及其对TGFβ-1/p38MAPK信号转导通路的影响[J].中华中医药杂志,2012,27(4):1092-1097,1220-1221.
[8]朱晓宾.早期糖尿病肾病临床诊断中尿肾功检测的应用价值探讨[J].中国医药指南,2017,15(2):86-87.
[9]汪容,曹和欣.炎症及氧化应激机制与糖尿病肾病关系[J].辽宁中医药大学学报,2016,18(5):98-101.
[10]吴冰.糖尿病肾病的发病机制和临床治疗现状[J].糖尿病新世界,2016(6):197-198.
[11]郭晓玲,康丽霞,檀金川.芪芍胶囊联合厄贝沙坦治疗糖尿病肾病[J].中国实验方剂学杂志,2016,22(3):183-187.
[12]林兰,郭小舟,倪青,等.早期糖尿病肾病的相关危险因素及中医证型分析[J].上海中医药大学学报,2010,24(1):29-31.
[13]孙超,谢晴宇,孟庆刚.糖尿病肾病中医证素分布规律研究[J].北京中医药大学学报,2015,38(4):266-270.
[14]何玉明,丁宪群.尿毒清颗粒对早中期糖尿病肾病患者炎症因子影响的临床研究[J].中药药理与临床,2015,31(6):160-162.
[15]王玉浔,安雅臣,赵晓晶,等.尿毒清颗粒对糖尿病肾病大鼠P选择素水平的影响[J].实用医学杂志,2013,29(15):2469-2471.
[16]陈琇萌,俞小敏,肖洁.尿毒清颗粒抑制大鼠肾脏间质纤维化作用靶点研究[J].湖南中医药大学学报,2017,37(1):33-37.
[17]朱蓓,宋卫国,彭璘.糖尿病肾病微炎症及氧化应激的中医药研究进展[J].江西中医药,2016,47(7):72-74.
[18]梁泽智,祝胜郎.p38MAPK及其在糖尿病肾病中的作用研究进展[J].医学综述,2015,16(19):3542-3544.
[2]李艳丽,廖勇敢,李晓雯,等.糖尿病肾病发病的危险因素分析[J].实用预防医学,2017,24(2):133-136.
[3]朱宜临,钱秋海,张新颖.糖尿病肾病中西医研究进展[J].山东中医杂志,2017,36(1):82-84,88.
[4]路文静,武士锋,杨洪涛.尿毒清颗粒治疗糖尿病肾病相关机理研究进展[J].河南中医,2017,37(1):181-184.
[5]张秀峰,张月恒.糖尿病肾病的临床护理方法以及体会[J].中国医药指南,2017,15(4):233-234.
[6]冼启经,曾莉.尿毒清胶囊对梗阻性肾病大鼠肾间质纤维化的影响[J].新中医,2011,43(3):128-130.
[7]乌格敦其其格,赵宗江,蒋玉凤,等.糖肾平胶囊对STZ诱导糖尿病肾病大鼠肾脏保护及其对TGFβ-1/p38MAPK信号转导通路的影响[J].中华中医药杂志,2012,27(4):1092-1097,1220-1221.
[8]朱晓宾.早期糖尿病肾病临床诊断中尿肾功检测的应用价值探讨[J].中国医药指南,2017,15(2):86-87.
[9]汪容,曹和欣.炎症及氧化应激机制与糖尿病肾病关系[J].辽宁中医药大学学报,2016,18(5):98-101.
[10]吴冰.糖尿病肾病的发病机制和临床治疗现状[J].糖尿病新世界,2016(6):197-198.
[11]郭晓玲,康丽霞,檀金川.芪芍胶囊联合厄贝沙坦治疗糖尿病肾病[J].中国实验方剂学杂志,2016,22(3):183-187.
[12]林兰,郭小舟,倪青,等.早期糖尿病肾病的相关危险因素及中医证型分析[J].上海中医药大学学报,2010,24(1):29-31.
[13]孙超,谢晴宇,孟庆刚.糖尿病肾病中医证素分布规律研究[J].北京中医药大学学报,2015,38(4):266-270.
[14]何玉明,丁宪群.尿毒清颗粒对早中期糖尿病肾病患者炎症因子影响的临床研究[J].中药药理与临床,2015,31(6):160-162.
[15]王玉浔,安雅臣,赵晓晶,等.尿毒清颗粒对糖尿病肾病大鼠P选择素水平的影响[J].实用医学杂志,2013,29(15):2469-2471.
[16]陈琇萌,俞小敏,肖洁.尿毒清颗粒抑制大鼠肾脏间质纤维化作用靶点研究[J].湖南中医药大学学报,2017,37(1):33-37.
[17]朱蓓,宋卫国,彭璘.糖尿病肾病微炎症及氧化应激的中医药研究进展[J].江西中医药,2016,47(7):72-74.
[18]梁泽智,祝胜郎.p38MAPK及其在糖尿病肾病中的作用研究进展[J].医学综述,2015,16(19):3542-3544.