摘要
目的研究秦香止泻汤对人轮状病毒(HRV)腹泻乳鼠小肠黏膜细胞诱导型一氧化氮合酶mRNA(iNOS mR-NA)表达的影响,探讨该方干预HRV腹泻的作用机制。方法 192只出生5 d的昆明乳鼠随机分成空白对照组、模型对照组、病毒唑阳性药物组、秦香止泻汤低、中、高剂量组,每组32只,空白组经口灌入不含HRV的培养液,其余各组经口灌入50μL(3.0×108PFU/mL)的Wa株HRV培养液。接种病毒12 h后,实验组每只乳鼠经口给予相应的秦香止泻汤药液50μL,阳性对照组给予同体积的病毒唑,空白对照组和模型对照组分别给予同体积的蒸馏水。连续给药8 d,胶体金试纸方法检测粪便HRV抗原,观察一般情况和腹泻评分,采用实时荧光定量PCR检测小肠黏膜iNOS mRNA的表达,并对腹泻评分及iNOS mRNA表达进行相关性分析。结果接种病毒后第2天开始出现不同程度的腹泻,秦香止泻汤组与模型组或病毒唑组比能明显缩短腹泻病程,减轻腹泻程度,呈现一定的量效关系,且效果优于病毒唑(P<0.05)。1～8 d模型对照组与空白对照组比,循环阈值(CT值)明显降低(P<0.01)。各药物组CT值与模型对照组比显著升高(P<0.05),其中秦香止泻汤高剂量组在第4天较低剂量组、模型组和阳性药物组明显升高(P<0.01);相关性分析显示iNOS mRNA表达CT值与腹泻评分呈负相关(k=-5.9156,R2=0.913)。结论秦香止泻汤能明显缩短HRV感染乳鼠腹泻病程,减轻腹泻程度,可能是通过调低小肠黏膜细胞iNOS mRNA的表达而发挥作用。
AIM To study the effect of Qinxiang Zhixie Decoction on inducible nitric oxide synthase（iNOS） mRNA expression of human rotavirus（HRV） diarrhea in neonatal rat and to discuss the intervention mechanism of HRV diarrhea.METHODS One hundred and ninety-two five-day-old Kunming rats were randomly divided into the negative control group,model group,virazole positive drug group and Qinxiang Zhixie Decoction group with low,moderate and high doses.There were thirty-two rats in each group.Rats in negative control group were fed with culture medium without HRV,while those in other groups were infused by mouth Wa strain HRV culture liquid of 50 μL（3.0×108 PFU/mL）.After 12 h of inoculation of the virus,each rat of the experimental group was fed with 50 μL of Qinxiang Zhixie Decoction.Rats in the positive control group were given the same amount of ribavirin.Those in negative control group and model group were given the same amount of distilled water.In eight days of continuous infusion,colloidal gold method was used for the detection of fecal HRV antigen,and observed general situation and diarrhea score.Fluorescence quantitative PCR was used to detect and calculate the expression of small intestinal mucosal iNOS mRNA expression and relevant correlation analysis.RESULTS Infected with virus,rats started the different levels diarrhea compared with model group or virazole group.Qinxiang Zhixie Decoction group obviously shortened diarrhea period,relieved diarrhea in a certain dose-dependent manner,and its effect was better than that of ribavirin（P0.05）.Compared with negative control group from the first to the eighth day,cycle threshold in model group was significantly lower（P0.01）.Compared with model group,cycle threshold in every drug group increased significantly（P0.05）.Cycle threshold in high dose of Qinxiang Zhixie Decoction group on the fourth day was obviously higher than that in low dose of Qinxiang Zhixie Decoction group,model group and positive drug group（P0.01）.Correlation analysis showed iNOS mRNA expression of cycle threshold value and diarrhea score was negative correlation（k=-5.915 6,R2=0.913）.CONCLUSION Qinxiang Zhixie Decoction can greatly shorten diarrhea period of neonatal rats with HRV infection and relieve diarrhea,and is probably related to the decrease of iNOS mRNA expression in the intestinal mucosal cell.
出处
《中成药》
CAS
CSCD
北大核心
2013年第4期647-651,共5页
Chinese Traditional Patent Medicine
基金
湖南省科学技术厅(2011FJ3025)
