葛根素对病毒性心肌炎的保护机制研究  

Protective effect of puerarin on viral myocarditis and the underlying mechanism

作　　者：张晓梅 孙景辉[2] 朱华 孟晓波 窦忠霞 Zhang Xiaomei;Sun Jinghui;Zhu Hua;Meng Xiaobo;Dou Zhongxia(Department of Pediatrics, the People's Hospital of the Inner Mongolia Autonomous Region, Hohhot 010017, China;Department of Pediatrics, the People's Hospital of the Inner Mongolia Autonomous Region, Hohhot 010017, China)

机构地区：[1]内蒙古自治区人民医院儿科,呼和浩特010017 [2]吉林大学白求恩第一医院小儿心血管科,长春130021

出　　处：《国际儿科学杂志》2019年第2期140-143,F0003,共5页International Journal of Pediatrics

摘　　要：目的探讨Nrf2在病毒性心肌炎小鼠中的表达及葛根素(Pue)治疗后Nrf2的变化及作用。方法将130只BALB/c纯种4周龄雄性小鼠,随机分成对照组(20只)、VMC组(30只)、Nrf2激活剂组(20只)、不同浓度Pue组(各20只),用柯萨奇B3型病毒(CVB3),通过VMC经典造模方法进行造模。分别在实验第0、4、7、14、28天采血、杀鼠,留取心肌组织。采用流式细胞术检测小鼠心肌细胞的凋亡情况,应用Real-TimePCR法检测Nrf2mRNA、HO-1mRNA、FasmRNA、TGF-β1mRNA的水平及用Westernblot法检测Nrf2、HO-1、Fas、TGF-β1的蛋白表达情况。应用统计软件SPSS19.0对结果进行统计学分析,计量资料以均值±标准差(±s)表示,配对样本用均值t检验,成组资料用双向方差分析,相关性分析应用Spearman相关检验,P<0.05为差异有显著性。结果Nrf2mRNA及其蛋白在各组均有表达。通过CPDT以及Pue分析Nrf2与HO-1、Fas、TGF-β1之间的相关关系,得出完全一致的结果,说明Nrf2与HO-1、Fas、TGF-β1之间的关系,不因干预措施的改变而变化。CPDT及Pue组HO-1的转录及蛋白表达明显升高,且与Nrf2呈正相关(r=0.969,P<0.01)。在一定Pue剂量梯度(≤45mg/kg)下,HO-1的转录及蛋白表达呈剂量依赖性;CPDT及Pue组心肌细胞凋亡数减少,Nrf2与Fas呈负相关(r=-0.968,P<0.01);CPDT及Pue组中TGF-β1在一定的剂量梯度下,随着剂量的增大,TGF-β1表达降低,Nrf2与TGF-β1呈负相关(r=-0.753,P<0.01)。结论Nrf2在VMC中表达增高,参与VMC的发生、发展过程。Nrf2在VMC中通过上调抗氧化酶HO-1达到抗氧化作用;通过抑制Fas/FasL信号通路实现抗心肌凋亡作用;通过抑制TGF-β1的转录及蛋白表达,抑制心肌纤维化。Pue对VMC的治疗作用是通过激活Nrf2,进而产生抗氧化、抗心肌凋亡和抗心肌纤维化作用。Objective To detect the expression of Nrf2 in mice with viral myocarditis and to investigate the changes and effects of Nrf2 after puerarin(Pue)treatment. MethodsA total of 130 BALB /C male mice aged 4 weeks were randomly divided into control group, VMC group, Nrf2 activator group and Pue group(20 mice in each group)with different concentrations.The models were made with Coxsackie B3 virus(CVB3). The mice were sacrificed on day 0, 4, 7, 14 and 28 respectively, and blood and myocardial samples were harvested.Cardiomyocyte apoptosis was detected by flow cytometry.The expression changes of Nrf2, HO-1, Fas, TGF-beta 1 mRNA were detected by real-time PCR and Western blot respectively.Statistical software SPSS19.0 was used to analyze the results.The measurement data was expressed mean ± standard deviation.The paired samples were tested with mean t test.The group data were analyzed with two-way ANOVA.A P value of less than 0.05 was considered to indicate statistical significance.Correlation analysis was performed with Spearman′s correlation test. ResultsNrf2 mRNA and Nrf2 protein were expressed in all groups.The correlations between Nrf2 and HO-1, Fas and TGF-beta-1 were analyzed according to CPDT or Pue, and the results were consistent with each other.It showed that the relationship between Nrf2 and HO-1, Fas and TGF-beta-1 did not change with intervention measures.The transcription and protein expression of HO-1 in CPDT and Pue groups were significantly increased, and were positively correlated with Nrf2(r=0.969, P<0.01). At a certain dose gradient(<45 mg/kg), the transcription and protein expression of HO-1 were dose-dependent;the decreased cardiomyocyte apoptosis was observed in both CPDT and Pue group, while Nrf2 and Fas were negatively correlated(r=-0.968, P<0.01);at a certain dose gradient, the expression of TGF-beta 1 in CPDT and Pue group decreased with the increase of dose, and Nrf2 and TGF-beta 1 were negatively correlated(r=-0.753, P<0.01). ConclusionThe increased expression of Nrf2 in VMC is involved in

关 键 词：病毒性心肌炎 BALB /c鼠 柯萨奇病毒B3 NRF2 葛根素 

分 类 号：R285.5[医药卫生—中药学]