黄芩苷体外抑制柯萨奇病毒B组3型感染的机制研究
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淮安市自然科学基金(HAB201722);南京医科大学校基金面上项目(2016NJMU137)


Study on the mechanism of baicalin against coxsackie virus B3 infection in vitro
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    摘要:

    目的:探讨黄芩苷(baicalin,BA)体外抑制柯萨奇病毒B组3型(coxsackie virus B3,CVB3)感染的效用及机制。方法:CellTiter 96- AQueous 单溶液试剂检测Hela细胞存活率,病毒空斑实验检测病毒滴度,CHOD-PAP法和ACS-ACOD法分别用于检测细胞中游离脂肪酸(free fatty acids,FAA)和胆固醇(cholesterol,CHO)的含量,RT-PCR检测脂类代谢关键分子脂肪酸合成酶(fatty acid synthase,Fasn)和乙酰-CoA羧化酶(acetyl-CoA carboxylase,ACC)的mRNA表达,Western blot检测CVB3病毒衣壳蛋白VP1表达。结果:BA在低于100 μg/mL时对Hela细胞无明显毒性效用,但能呈剂量依赖地抵抗CVB3诱导细胞病变,显著减少病毒滴度和抑制CVB3/VP1的表达;CVB3感染能显著增加细胞中FFA和CHO含量以及Fasn和ACC的mRNA表达,但是经BA处理后,能显著减少这些脂类代谢物质的含量;棕榈酸预处理细胞,能恢复BA处理组中CVB3/VP1的表达。结论:BA能通过调节细胞脂类合成来发挥抗CVB3感染的效用。

    Abstract:

    Objective:To investigate the efficacy and mechanism of baicalin(BA)inhibiting coxsackie virus B3(CVB3) infection in vitro. Methods:Hela cell viability was measured by CellTiter 96- AQueous single reagent. The virus titer was detected by virus plaque assay. The concentration of cholesterol(CHO)and free fatty acids(FAA)were detected by CHOD-PAP and ACS-ACOD,respectively. The expression levels of fatty acid synthase(Fasn) mRNA and acetyl-CoA carboxylase(ACC) mRNA were detected by RT-PCR. The expression of VP1 protein was detected by Western blot in CVB3 virus. Results:BA had no significant toxic effect on Hela cells at less than 100 μg/mL,but was able to against cytopathy by CVB3 in a dose-dependent manner. The viral titers and CVB3/VP1 expression were significantly reduced by BA treatment in CVB3 infected cells. The levels of CHO and FAA,mRNA of Fasn and ACC were all significantly expressed in CVB3 infected cells,but ware all significantly reduced by BA treatment. Compared with BA treatment group by BSA pretreatment,palmitic acid pretreatment could increase the expression of CVB3/VP1 in Hela cells. Conclusion:BA can against CVB3 infection by regulating cellular lipid synthesis.

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金 跃,李 祥,王晓丽,马 莉,程建平,邢 飞,刘媛媛.黄芩苷体外抑制柯萨奇病毒B组3型感染的机制研究[J].南京医科大学学报(自然科学版),2019,(2):196-200

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  • 收稿日期:2017-10-27
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  • 在线发布日期: 2019-03-06
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