电针联合两色金鸡菊提取物对糖尿病肾病大鼠肾脏Vimentin、α-SMA、TGF-β1及p-smad2表达的影响
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摘要
目的观察电针联合两色金鸡菊提取物对糖尿病大鼠肾脏波形蛋白(Vimentin)、α-平滑肌肌动蛋白(α-SMA)、转化生长因子-β1(TGF-β1)及p-smad2表达的影响,探讨其作用机制。方法将36只SD大鼠随机分为正常组、模型组、电针组、两色金鸡菊组、电针+两色金鸡菊组和二甲双胍组,每组6只。采用高糖高脂喂养联合腹腔注射链脲佐菌素建立2型糖尿病模型。正常组、模型组不进行干预;电针组针刺大鼠双侧"肾俞""脾俞",连接电针,留针30 min;两色金鸡菊组予两色金鸡菊乙酸乙酯提取物(300 mg/kg)灌胃;电针+两色金鸡菊组予电针联合两色金鸡菊乙酸乙酯提取物灌胃;二甲双胍组予二甲双胍(200mg/kg)药液灌胃。每日1次,治疗4周。HE染色观察肾组织形态学及纤维化程度,Western blot检测肾组织Vimentin、α-SMA、TGF-β1及p-smad2蛋白表达。结果 HE染色结果显示,电针组、两色金鸡菊组、电针+两色金鸡菊组和二甲双胍组均可缓解模型大鼠肾小管扩张,减轻肾小球皱缩现象,改善炎性细胞浸润,减少基底膜增生和融合,且以电针+两色金鸡菊组改善更明显;Westernblot检测结果显示,电针和两色金鸡菊提取物均可降低模型大鼠肾组织Vimentin、α-SMA、TGF-β1及p-smad2蛋白表达,且以两者联合效果最佳。结论电针联合两色金鸡菊提取物可明显改善糖尿病大鼠肾脏纤维化,其作用机制可能与抑制TGF-β1/Smad2信号通路有关。
Objective To observe the effects of electroacupuncture combined with coreopsis tinctoria extract for expressions of Vimentin, α-SMA, TGF-β1 and p-smad2 in renal tissue of diabetic rats; To discuss its mechanism of action. Methods Totally thirty-six SD rats were randomly divided into normal group, model group,electroacupuncture group, coreopsis tinctoria group, electroacupuncture combined with coreopsis tinctoria group and metformin group, with 6 rats in each group. Type 2 DM model was established by high-fat and high-glucose diet and STZ injection intraperitoneally. Normal group and model group were given no treatment; Electroacupuncture was at the acupoints of "Pishu" and "Shenshu" once daily in electroacupuncture group for 30 min; coreopsis tinctoria group was given intragastric administration of 300 mg/kg coreopsis tinctoria extract for gavage; electroacupuncture combined with coreopsis tinctoria group was given electroacupuncture and intragastric administration of coreopsis tinctoria extract for gavage; metformin group was given intragastric administration of 200 mg/kg metformin for gavage. Histological changes and fibrosis degree of kidneys were observed by HE staining and expressions of Vimentin, α-SMA, TGF-β1 and p-smad2 were detected by Western blot. Results HE staining results indicated that electroacupuncture, coreopsis tinctoria, electroacupuncture combined with coreopsis tinctoria and metformin groups could relieve the dilation of renal tubules, reduce the phenomenon of glomerular shrinkage, improve the infiltration of inflammatory cells and reduce the hyperplasia and fusion of basal membranes. Furthermore, electroacupuncture combined with coreopsis tinctoria group performed best. Western blot analysis revealed that both electroacupuncture and coreopsis tinctoria extract could reduce the expressions of Vimentin, α-SMA, TGF-β1 and p-smad2 while combination work better. Conclusion Electroacupuncture combined with coreopsis tinctoria extract can improve renal fibrosis in diabetic rats. The mechanism may be related with the inhibition of TGF-β1/smad2 pathway.
Objective To observe the effects of electroacupuncture combined with coreopsis tinctoria extract for expressions of Vimentin, α-SMA, TGF-β1 and p-smad2 in renal tissue of diabetic rats; To discuss its mechanism of action. Methods Totally thirty-six SD rats were randomly divided into normal group, model group,electroacupuncture group, coreopsis tinctoria group, electroacupuncture combined with coreopsis tinctoria group and metformin group, with 6 rats in each group. Type 2 DM model was established by high-fat and high-glucose diet and STZ injection intraperitoneally. Normal group and model group were given no treatment; Electroacupuncture was at the acupoints of "Pishu" and "Shenshu" once daily in electroacupuncture group for 30 min; coreopsis tinctoria group was given intragastric administration of 300 mg/kg coreopsis tinctoria extract for gavage; electroacupuncture combined with coreopsis tinctoria group was given electroacupuncture and intragastric administration of coreopsis tinctoria extract for gavage; metformin group was given intragastric administration of 200 mg/kg metformin for gavage. Histological changes and fibrosis degree of kidneys were observed by HE staining and expressions of Vimentin, α-SMA, TGF-β1 and p-smad2 were detected by Western blot. Results HE staining results indicated that electroacupuncture, coreopsis tinctoria, electroacupuncture combined with coreopsis tinctoria and metformin groups could relieve the dilation of renal tubules, reduce the phenomenon of glomerular shrinkage, improve the infiltration of inflammatory cells and reduce the hyperplasia and fusion of basal membranes. Furthermore, electroacupuncture combined with coreopsis tinctoria group performed best. Western blot analysis revealed that both electroacupuncture and coreopsis tinctoria extract could reduce the expressions of Vimentin, α-SMA, TGF-β1 and p-smad2 while combination work better. Conclusion Electroacupuncture combined with coreopsis tinctoria extract can improve renal fibrosis in diabetic rats. The mechanism may be related with the inhibition of TGF-β1/smad2 pathway.
引文
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[9]韩华林,赵旭红.糖尿病肾病患者采取补脾益肾汤配合针灸治疗的临床疗效[J].世界最新医学信息文摘,2018,18(70):23-24.
[10]许碧香,黎亚玲,赵若华.基于文献分析糖尿病肾病临床用穴规律[J].中医药临床杂志,2017,29(10):1692-1696.
[11]NIE Y,LI S,YI Y,et al.Effects of astragalus injection on the TGF-β/Smad pathway in the kidney in type 2 diabetic mice[J].BMC Complementary&Alternative Medicine,2014,14(1):1-8.
[12]VOELKER J,BERG P H,SHEETZ M,et al.Anti-TGF-β1 antibody therapy in patients with diabetic nephropathy[J].Journal of the American Society of Nephrology,2017,28(3):953.
[13]AL-ONAZI A S,AL-RASHEED N M,ATTIA H A,et al.Ruboxistaurin attenuates diabetic nephropathy via modulation of TGF-β1/Smad and GRAP pathways[J].Journal of Pharmacy&Pharmacology,2016,68(2):219-232.
[14]ZHANG Y,WANG S,LIU S,et al.Role of Smad signaling in kidney disease[J].International Urology&Nephrology,2015,47(12):1965-1975.
[15]WU J S,SHI R,LU X,et al.Combination of active components of Xiexin decoction ameliorates renal fibrosis through the inhibition of NF-κB and TGF-β1/Smad pathways in db/db diabetic mice[J].PLoS One,2015,10(3):e0122661.
[16]张楠楠,马瑶,邓凯,等.两色金鸡菊黄酮提取物通过对高糖高脂诱导的大鼠肾小球系膜细胞纤维化的影响及相关机制研究[J].中药材,2018,41(4):950-955.
[2]姚蓝,张雨洁,刘菁,等.两色金鸡菊提取物对大鼠肾小球系膜细胞增殖及纤维化因子表达的影响[J].中国实验方剂学杂志,2017,23(19):134-139.
[3]姚蓝,蒋洁,王靖宣,等.两色金鸡菊对糖尿病大鼠肾脏纤维化Rho A/ROCK/Nox4信号通路的影响[J].中国中医药信息杂志,2019,26(1):57-62.
[4]何文娟,陈丽,舒晴,等.“标本配穴”电针对糖尿病肾病的改善作用及肾脏SIRT1蛋白表达水平的影响[J].中国中医基础医学杂志,2017,24(9):1280-1283.
[5]张雨洁,兰怡,李卉,等.两色金鸡菊乙酸乙酯提取物对STZ诱导的糖尿病SD大鼠的影响[J].中国药理学通报,2015,31(10):1439-1443.
[6]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:316.
[7]李承德,王煜,曲敬蓉,等.黄芪多糖对糖尿病大鼠肾脏TGF-β1/Smads信号通路的影响[J].中国药理学通报,2018,34(4):512-516.
[8]李志杰,张悦,陆海英,等.六味地黄丸对糖尿病肾病大鼠肾组织TGF-β1-smad通路的影响[J].时珍国医国药,2017,28(8):1811-1814.
[9]韩华林,赵旭红.糖尿病肾病患者采取补脾益肾汤配合针灸治疗的临床疗效[J].世界最新医学信息文摘,2018,18(70):23-24.
[10]许碧香,黎亚玲,赵若华.基于文献分析糖尿病肾病临床用穴规律[J].中医药临床杂志,2017,29(10):1692-1696.
[11]NIE Y,LI S,YI Y,et al.Effects of astragalus injection on the TGF-β/Smad pathway in the kidney in type 2 diabetic mice[J].BMC Complementary&Alternative Medicine,2014,14(1):1-8.
[12]VOELKER J,BERG P H,SHEETZ M,et al.Anti-TGF-β1 antibody therapy in patients with diabetic nephropathy[J].Journal of the American Society of Nephrology,2017,28(3):953.
[13]AL-ONAZI A S,AL-RASHEED N M,ATTIA H A,et al.Ruboxistaurin attenuates diabetic nephropathy via modulation of TGF-β1/Smad and GRAP pathways[J].Journal of Pharmacy&Pharmacology,2016,68(2):219-232.
[14]ZHANG Y,WANG S,LIU S,et al.Role of Smad signaling in kidney disease[J].International Urology&Nephrology,2015,47(12):1965-1975.
[15]WU J S,SHI R,LU X,et al.Combination of active components of Xiexin decoction ameliorates renal fibrosis through the inhibition of NF-κB and TGF-β1/Smad pathways in db/db diabetic mice[J].PLoS One,2015,10(3):e0122661.
[16]张楠楠,马瑶,邓凯,等.两色金鸡菊黄酮提取物通过对高糖高脂诱导的大鼠肾小球系膜细胞纤维化的影响及相关机制研究[J].中药材,2018,41(4):950-955.