蜘蛛香不同提取物抗轮状病毒的作用研究  

Study on the anti-rotavirus effect of different extracts of Valeriana jatamansi Jones

作　　者：李峰 宋丽军 周玉晶 冯宇璇 袁月 钱余培 杨思雁 易灏森 杨芷胭 余润宇 赵文昌 LI Feng;SONG Li-jun;ZHOU Yu-jing;FENG Yu-xuan;YUAN Yue;QIAN Yu-pei;YANG Si-yan;YI Haosen;YANG Zhi-yan;YU Run-yu;ZHAO Wen-chang(School of pharmacy,Guangdong Medical University,Dongguan 523808,China;Dongguan Key Laboratory of Screening and Research of Anti-inflammatory Ingredients in Chinese Medicine,Dongguan 523808,China;Guangdong Key Laboratory for Research and Development of Natural Drugs,Guangdong Medical University,Zhanjiang 524023,China)

机构地区：[1]广东医科大学药学院 [2]东莞市中药抗炎活性成分筛选与研究重点实验室,广东东莞523808 [3]广东医科大学天然药物研究开发重点实验室,广东湛江524023

出　　处：《广东医科大学学报》2024年第4期356-364,共9页Journal of Guangdong Medical University

基　　金：国家自然科学基金面上项目(81473401,81973548),广东省高校创新团队项目(2022KCXTD011),广东医科大学学科重点项目(4SG22009G),湛江市科技计划项目(2021B01139)。

摘　　要：目的研究蜘蛛香不同提取物抗轮状病毒(Rotavirus,RV)的作用。方法采用RV-Wa株感染MA104细胞模型,MTT染色法测定病毒抑制率,考察蜘蛛香水提物、70%乙醇提取物(ZZX-1)及90%乙醇提取后经大孔吸附树脂法纯化所得提取物(ZZX-2)体外抗RV的作用;采用免疫荧光和实时荧光定量PCR(qRT-PCR)检测RV结构蛋白VP6的表达变化,验证蜘蛛香抗RV作用;采用RV-Wa株感染Caco-2细胞,qRT-PCR检测蜘蛛香对神经营养因子GDNF、紧密连接蛋白ZO-1和促炎因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)]的影响。结果蜘蛛香水提物无抗RV的作用;蜘蛛香醇提物ZZX-1和ZZX-2具有抗RV生物合成作用。免疫荧光和qRT-PCR实验结果表明,ZZX-1和ZZX-2均能够抑制VP6蛋白的合成以及降低VP6 mRNA的表达,抑制RV的合成。与RV组相比,ZZX-1和ZZX-2能够显著提高GDNF、ZO-1 mRNA的相对表达量(P<0.001或0.01),降低TNF-α、IL-1β、IL-6 mRNA的相对表达量(P<0.0001或0.001)。结论蜘蛛香醇提物ZZX-1和ZZX-2具有抗RV生物合成作用,可能通过提高GDNF、ZO-1基因表达,降低炎症因子IL-1β、IL-6、TNF-α的基因表达发挥抗RV感染作用。Objective The aim of this study is to examine the anti-rotavirus(RV)effects of different extracts of Valeriana jatamansi Jones.Methods The RV-Wa strain infected MA104 cell model was used,and the virus inhibition rate was determined by using MTT staining.The in vitro anti-RV activity of spider perfume extract,70%ethanol extract(ZZX-1)and 90%ethanol extract(ZZX-2)was purified by the macroporous adsorption resin method.Immunofluorescence and real-time fluorescence quantitative PCR(qRT-PCR)were used to detect the expression of RV structural protein VP6,and to verify the anti-RV effect of spide.Caco-2 cells were infected with RV-Wa strain and the effects of arachnid on the expressions of neurotrophic factor GDNF,tight junction protein ZO-1 and pro-inflammatory factors(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6)were detected by qRT-PCR.Results The extract of Valeriana jatamansi Jones did not have any anti-RV effect;Valeriana jatamansi Jones extracts ZZX-1 and ZZX-2 have anti-RV biosynthesis effects.The results of immunofluorescence and qRT-PCR indicate that both ZZX-1 and ZZX-2 can reduce the fluorescence and mRNA expression of VP6,and inhibit RV synthesis.Compared with the RV group,ZZX-1 and ZZX-2 significantly increased the expression levels of neurotrophic factor GDNF and tight junction protein ZO-1 mRNA(P<0.001 or 0.01),and reduced the relative expression levels of tumor necrosis factor alpha(TNF alpha),interleukin-1β(IL-1β),and interleukin-6(IL-6)mRNA(P<0.0001 or 0.001).Conclusion The alcohol extracts ZZX-1 and ZZX-2 from the traditional Chinese medicine Valeriana jatamansi Jones have anti-RV biosynthesis effects,which may exert anti-RV infection effects by increasing the expression of GDNF and ZO-1 and reducing the expression of inflammatory factors IL-1β,IL-6,and TNF-α.

关 键 词：轮状病毒 蜘蛛香 环烯醚萜类 肠上皮细胞 紧密连接蛋白 

分 类 号：R284[医药卫生—中药学]