通心络胶囊联合津力达颗粒对糖尿病肾病大鼠TGF-β1/Smad信号转导通路的作用研究
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摘要
目的:观察通心络胶囊联合津力达颗粒对糖尿病肾病(DN)大鼠TGF-β1/Smad信号转导通路的作用。方法:将80只SD大鼠分为正常组(N组)、模型组(M组)、厄贝沙坦组(I组)、通心络组(T组)、津力达组(J组)、通心络加津力达组(TJ组);以高糖高脂饲料喂养加尾静脉注射链脲佐菌素建立糖尿病动物模型;采用免疫组化SP法检测Ⅳ型胶原的表达;以Western Blot法检测TGF-β1/Smad信号转导通路中Smad2/3、p-Smad2/3及Smad7的表达水平。结果:Ⅳ型胶原在M组中表达增强,在I组与TJ组中均有减弱,其中TJ组的表达强度及范围更接近N组。与N组相比,M组中Smad2/3与p-Smad2/3蛋白表达显著增强(P<0.01,P<0.05),而Smad7表达则显著减弱(P<0.01);与M组相比,I组与TJ组p-Smad2/3蛋白表达显著减弱(P<0.05),Smad7蛋白表达显著增强(P<0.01)。结论:通心络胶囊联合津力达颗粒可抑制TGF-β1/Smad信号转导通路并减弱Ⅳ型胶原的表达,这可能是其治疗DN的机制之一。
Objective: To observe the effects of Tongxinluo Capsule combined with Jinlida Granules on transforming growth factor(TGF)-β1/Smad signal transduction pathway in rats with diabetic nephropathy(CON). Methods: A total of 80 SD rats were randomly divided into normal group(N), model group(M), irbesartan group(I), Tonxinluo group(T), Jinlida group(J) and combined group of Tonxinluo and Jinlida(TJ). Animal model of diabetes were established by intravenous injecting in tails with streptozotocin(STZ), the expression of type Ⅳ collagen were detected with immunohistochemical assay of SP; Western Blot was used to detect the expression levels of Smad2/3, p-Smad2/3 and Smad7 in TGF-β1/Smad signal transduction pathways. Results:The expression of type Ⅳ collagen was significantly increased in M group and decreased in Ⅰ group and TJ group, and the expression of type Ⅳ collagen in TJ group were similar with that in N group. Compared with N group, the expression of Smad2/3 and p-Smad2/3 protein was significantly increased in M group(P<0.01, P<0.05), while the expression of Smad7 wasdecreased in M group(P<0.01); Compared with M group, the expression of p-Smad2/3 protein was significantly decreased in I group and TJ group(P<0.05), and the expression of Smad7 was increased(P<0.01). Conclusion: Tongxinluo Capsule combined with Jinlida Granules can inhibit TGF-β1/Smad signal transduction pathway and reduce the expression of type Ⅳ collagen, which may be one of mechanisms of treatment to DN.
Objective: To observe the effects of Tongxinluo Capsule combined with Jinlida Granules on transforming growth factor(TGF)-β1/Smad signal transduction pathway in rats with diabetic nephropathy(CON). Methods: A total of 80 SD rats were randomly divided into normal group(N), model group(M), irbesartan group(I), Tonxinluo group(T), Jinlida group(J) and combined group of Tonxinluo and Jinlida(TJ). Animal model of diabetes were established by intravenous injecting in tails with streptozotocin(STZ), the expression of type Ⅳ collagen were detected with immunohistochemical assay of SP; Western Blot was used to detect the expression levels of Smad2/3, p-Smad2/3 and Smad7 in TGF-β1/Smad signal transduction pathways. Results:The expression of type Ⅳ collagen was significantly increased in M group and decreased in Ⅰ group and TJ group, and the expression of type Ⅳ collagen in TJ group were similar with that in N group. Compared with N group, the expression of Smad2/3 and p-Smad2/3 protein was significantly increased in M group(P<0.01, P<0.05), while the expression of Smad7 wasdecreased in M group(P<0.01); Compared with M group, the expression of p-Smad2/3 protein was significantly decreased in I group and TJ group(P<0.05), and the expression of Smad7 was increased(P<0.01). Conclusion: Tongxinluo Capsule combined with Jinlida Granules can inhibit TGF-β1/Smad signal transduction pathway and reduce the expression of type Ⅳ collagen, which may be one of mechanisms of treatment to DN.
引文
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[11]何笑云,于萍,欧春麟,等.高糖高溶血磷脂胆碱对系膜细胞产生纤维连接蛋白、Ⅳ型胶原及转化生长因子-β1的影响.中国糖尿病杂志,2016,24(2):156-158
[12]胡筱娟,秦艳,刘恬园,等.糖尿病肾病中医证型与肾络瘀阻相关性临床探讨.辽宁中医杂志,2016,43(1):91-92
[13]薛青.益肾泄浊方治疗糖尿病肾病气阴两虚、肾络瘀阻证临床研究.中医学报,2017,32(2):213-217
[14]胡筱娟,秦艳,刘恬园.基于肾络瘀阻病机与糖尿病肾病相关性临床研究.陕西中医,2015,36(12):1614-1616
[15]龙轩,王锋,黄昶荃.通心络胶囊治疗糖尿病肾病的系统评价.中国循证医学杂志,2010,10(1):73-80
[16]史婧丽,吴莹,宋玉萍,等.津力达颗粒对糖尿病大鼠胰岛B细胞的保护作用.第二军医大学学报,2012,33(4):385-389
[17]丁英钧,陈亮,王海蓉,等.通心络超微粉联合津力达颗粒对糖尿病大鼠肾功能及病理形态的影响.中药药理与临床,2014,30(5):112-115
[2]杜彦侠,陈亮,丁英钧,等.津力达颗粒联合通心络胶囊治疗糖尿病肾病的疗效观察.疑难病杂志,2012,11(6):425-427
[3]庞宗然,李晓军,刘祖涵,等.2型糖尿病合并大血管病变动物模型的建立和评价.中国中医基础医学杂志,2009,15(9):656-658
[4]童楠,张宁.中药益肾颗粒通过PI3k/Akt/m TOR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
[5]MassaguéJ.TGF-βsignal transduction.Annual Review of Biochemistry,2003,67(67):753-791
[6]Tesch G H,Lim A K.Recent insights into diabetic renal injury from the db/db mouse model of type 2 diabetic nephropathy.Am JPhysiol Renal Physiol,2011,300(2):F301
[7]Meng X M,Chung A C K,Lan H Y.Role of the TGF-β/BMP-7/Smad pathways in renal diseases.Clinical Science,2013,124(4):243
[8]Ka S M,Yeh Y C,Huang X R,et al.Kidney-targeting Smad7 gene transfer inhibits renal TGF-β/MAD homologue(SMAD)and nuclear factorκB(NF-κB)signalling pathways,and improves diabetic nephropathy in mice.Diabetologia,2012,55(2):509-519
[9]Mahendran K B,Bhaskar M V,Santha K,et al.Plasma and urinary typeⅣcollagen levels for early detection of nephropathy in type 2 diabetes mellitus patients.Int J Health Sci(Qassim),2016,10(4):492-498
[10]钟艳.C肽调控Ⅳ型胶原表达逆转糖尿病肾病纤维化的分子机制.石家庄:河北医科大学,2014
[11]何笑云,于萍,欧春麟,等.高糖高溶血磷脂胆碱对系膜细胞产生纤维连接蛋白、Ⅳ型胶原及转化生长因子-β1的影响.中国糖尿病杂志,2016,24(2):156-158
[12]胡筱娟,秦艳,刘恬园,等.糖尿病肾病中医证型与肾络瘀阻相关性临床探讨.辽宁中医杂志,2016,43(1):91-92
[13]薛青.益肾泄浊方治疗糖尿病肾病气阴两虚、肾络瘀阻证临床研究.中医学报,2017,32(2):213-217
[14]胡筱娟,秦艳,刘恬园.基于肾络瘀阻病机与糖尿病肾病相关性临床研究.陕西中医,2015,36(12):1614-1616
[15]龙轩,王锋,黄昶荃.通心络胶囊治疗糖尿病肾病的系统评价.中国循证医学杂志,2010,10(1):73-80
[16]史婧丽,吴莹,宋玉萍,等.津力达颗粒对糖尿病大鼠胰岛B细胞的保护作用.第二军医大学学报,2012,33(4):385-389
[17]丁英钧,陈亮,王海蓉,等.通心络超微粉联合津力达颗粒对糖尿病大鼠肾功能及病理形态的影响.中药药理与临床,2014,30(5):112-115