益气化瘀清热方及其拆方对大鼠肾系膜细胞表达NF-κBp65 mRNA及其蛋白的影响
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摘要
目的:把益气化瘀清热方拆方成益气、化瘀、清热3组,并与复方组和雷公藤组(TW组)对比,通过体外培养的方法,观察各组含药血清对大鼠肾小球系膜细胞(mesangial cells,MsC)中NF-κBp65 mRNA及蛋白的表达水平高低,进而探讨益气化瘀清热方的作用机制及不同类中药的作用强度。方法:采用RT-PCR法及Wersten Blot法分别检测含药血清对MsC表达NF-κBp65 mRNA及蛋白的作用强度。结果:各组含药血清均能抑制MsC中NF-κBp65 mRNA及其蛋白的表达,化瘀组、TW组作用较强(P<0.05);拆方后的益气、化瘀、清热3组比较:化瘀组作用较益气组、清热组明显增强(P<0.05)。结论:益气化瘀清热方及其拆方、TW均能抑制体外培养的大鼠MsC表达NF-κBp65mRNA及其蛋白,其中化瘀组、TW组作用较强,益气组较弱。
Objective:The paper is to divide the Yiqi Huayu Qingre Decoction into three groups named Yiqi,Huayu and Qingre,comparing complex prescription group and Tripterygium wilfordii group,and then through the method of culture in vitro so as to observe the effects of drug serum in each group both on the expressions of mRNA and protein of NF-κBp65,and further explore the strength of different medicines and the mechanism of Yiqi Huayu Qingre Decoction. Methods: We used the method of RT-PCR to detect the effect of medicine serum containing in each group on the expressions of mRNA of NF-κBp65. The effect of NF-κBp65 protein serum on the expression of MsC would be detected through Wersten Blot method. Results:The results showed that all containing medicine serum can restrain the expressions of mRNA and protein of NF-κBp65. The effects of containing medicine serum in Tripterygium wilfordi group and Huayu group were much stronger( P < 0. 05). Compared with these three discomposed groups of Yiqi,Huayu and Qingre,the effect of Huayu group was stronger than that of Yiqi group and Qingre group( P < 0. 05). Conclusion: Yiqi Huyu Qingre Decoction and its separated prescriptions and Tripterygium wilfordi can restrain the expressions of NF-κBp65 of the MsC cultured in vitro and its protein. The effect of Huayu serum group and Tripterygium wilfordii serum group is the strongest,and Yiqi serum group's is a little weaker.
Objective:The paper is to divide the Yiqi Huayu Qingre Decoction into three groups named Yiqi,Huayu and Qingre,comparing complex prescription group and Tripterygium wilfordii group,and then through the method of culture in vitro so as to observe the effects of drug serum in each group both on the expressions of mRNA and protein of NF-κBp65,and further explore the strength of different medicines and the mechanism of Yiqi Huayu Qingre Decoction. Methods: We used the method of RT-PCR to detect the effect of medicine serum containing in each group on the expressions of mRNA of NF-κBp65. The effect of NF-κBp65 protein serum on the expression of MsC would be detected through Wersten Blot method. Results:The results showed that all containing medicine serum can restrain the expressions of mRNA and protein of NF-κBp65. The effects of containing medicine serum in Tripterygium wilfordi group and Huayu group were much stronger( P < 0. 05). Compared with these three discomposed groups of Yiqi,Huayu and Qingre,the effect of Huayu group was stronger than that of Yiqi group and Qingre group( P < 0. 05). Conclusion: Yiqi Huyu Qingre Decoction and its separated prescriptions and Tripterygium wilfordi can restrain the expressions of NF-κBp65 of the MsC cultured in vitro and its protein. The effect of Huayu serum group and Tripterygium wilfordii serum group is the strongest,and Yiqi serum group's is a little weaker.
引文
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[8]姚源璋,张敏,曹鹏.清热解毒中药对TNF-α诱导的小鼠肾小球系膜细胞NF-κB信号转导通路的影响[J].中国中西医结合肾病杂志,2013,14(12):1047.
[9]熊涛,董文斌,王明勇,等.黄芪注射液对新生儿窒息后血清诱导人肾小管细胞损伤信号机制的影响[J].中国中西医结合杂志,2010,30(4):398-402.
[10]刘迟,郭刚,胡仲仪.莪术对单侧输尿管梗阻大鼠肾间质纤维化的影响[J].上海中医药杂志,2006,40(12):71-73.
[11]胡营杰.黄芪、水蛭有效组分对大鼠系膜细胞NF-κB、IκB、PDGF-BB、MMP-9的影响[D].南京:南京中医药大学,2016.
[12]彭卫华,南丽红,赖文芳,等.筋骨草总黄酮治疗系膜增生性肾小球肾炎大鼠肾组织NF-κB变化的实验研究[J].中华临床医师杂志(电子版),2013,7(32):11752-11757.
[13]程森华,陈安,方朝晖,等.丹蛭降糖胶囊对早期糖尿病肾病患者血NF-κB和尿微量蛋白排泄率及中远期疗效的影响[J].中华中医药学刊,2014,32(10):2338.
[14]费梅,熊佩华,孙美娟,等.清利活血养阴方对Ig A肾病患者血管内皮生长因子水平的影响[J].南京中医药大学学报,2012,28(4):327.
[15]陈睨,何立群.健脾清化方对肾纤维化大鼠肾功能、蛋白尿及肾组织Col-Ⅳ表达的影响[J].南京中医药大学学报,2013,29(6):548.
[16]王亿平,程敏,王东,等.清肾颗粒对单侧输尿管梗阻模型大鼠核转录因子-κB信号通路的干预作用[J].中国中西医结合急救杂志,2016,23(6):628-631.
[17]翟文生,杨濛,李广,等.益气化瘀清热方及其拆方影响体外培养大鼠Ms C增殖、产生Col-Ⅰ的实验研究[J].时珍国医国药,2015,26(2):499.
[18]翟文生,袁泉,李广,等.益气化瘀清热方及其拆方对体外培养大鼠系膜细胞产生Ⅲ型胶原蛋白的影响[J].中华中医药杂志,2015,30(9):3378.
[2]李健,杜芳,齐义军,等.食管癌及癌前病变中NF-κBp65/p50蛋白的表达[J].临床与实验病理学杂志,2006,22(5):560.
[3]Guijarro C,Kjm Y,Kasiske B L,et a1.Central role of the trascription factor NF-κB in mesangial cell production of chemokines[J].Contrib Nephrol,1997,120:210.
[4]康朋朋,张玉军,张景坤,等.狼疮性肾炎肾组织中NF-κB p65的表达及与细胞增殖的关系[J].临床与实验病理学杂志,2013,29(12):1332.
[5]丁桂霞,陈荣华,张爱华,等.膜增殖性肾炎核因子活化及其与系膜基质扩大的关系[J].南京医科大学学报,2001,21(3):192-195.
[6]LOPEZ-FRANCO O,SUZUKI Y,SANJUAN G,et al.Nuclear factor-kappa B inhibitors as potential novel anti-inflammatory agentsfor the treatment of immune glomerulonephritis[J].American Journal of Pathology,2002,161(4):1497-1505.
[7]翟文生,杨濛,张霞,等.益气化瘀清热方及其拆方对大鼠肾系膜细胞表达Smad2及泛素mRNA的影响[J].时珍国医国药,2017,28(5):1047.
[8]姚源璋,张敏,曹鹏.清热解毒中药对TNF-α诱导的小鼠肾小球系膜细胞NF-κB信号转导通路的影响[J].中国中西医结合肾病杂志,2013,14(12):1047.
[9]熊涛,董文斌,王明勇,等.黄芪注射液对新生儿窒息后血清诱导人肾小管细胞损伤信号机制的影响[J].中国中西医结合杂志,2010,30(4):398-402.
[10]刘迟,郭刚,胡仲仪.莪术对单侧输尿管梗阻大鼠肾间质纤维化的影响[J].上海中医药杂志,2006,40(12):71-73.
[11]胡营杰.黄芪、水蛭有效组分对大鼠系膜细胞NF-κB、IκB、PDGF-BB、MMP-9的影响[D].南京:南京中医药大学,2016.
[12]彭卫华,南丽红,赖文芳,等.筋骨草总黄酮治疗系膜增生性肾小球肾炎大鼠肾组织NF-κB变化的实验研究[J].中华临床医师杂志(电子版),2013,7(32):11752-11757.
[13]程森华,陈安,方朝晖,等.丹蛭降糖胶囊对早期糖尿病肾病患者血NF-κB和尿微量蛋白排泄率及中远期疗效的影响[J].中华中医药学刊,2014,32(10):2338.
[14]费梅,熊佩华,孙美娟,等.清利活血养阴方对Ig A肾病患者血管内皮生长因子水平的影响[J].南京中医药大学学报,2012,28(4):327.
[15]陈睨,何立群.健脾清化方对肾纤维化大鼠肾功能、蛋白尿及肾组织Col-Ⅳ表达的影响[J].南京中医药大学学报,2013,29(6):548.
[16]王亿平,程敏,王东,等.清肾颗粒对单侧输尿管梗阻模型大鼠核转录因子-κB信号通路的干预作用[J].中国中西医结合急救杂志,2016,23(6):628-631.
[17]翟文生,杨濛,李广,等.益气化瘀清热方及其拆方影响体外培养大鼠Ms C增殖、产生Col-Ⅰ的实验研究[J].时珍国医国药,2015,26(2):499.
[18]翟文生,袁泉,李广,等.益气化瘀清热方及其拆方对体外培养大鼠系膜细胞产生Ⅲ型胶原蛋白的影响[J].中华中医药杂志,2015,30(9):3378.