川楝素对柯萨奇病毒A16型体外抑制效应的研究

In vitro inhibitory effect of Toosendanin on coxsackie virus A16

目的探究川楝子提取物川楝素对柯萨奇病毒A16型(CA16)复制的抑制作用,及其相关的作用机制。方法将HCT116细胞分成空白组、阳性对照组与实验组。实验组用50μg·mL^(-1)川楝素培养CA16感染细胞,阳性对照组用24.4μg·mL^(-1)利巴韦林培养CA16感染细胞,空白组给予正常培养。用CCK-8实验检测川楝素对CA16感染的HCT116细胞活性的影响,用病毒空斑实验检测CA16的感染和复制能力,用蛋白质印迹法检测病毒结构蛋白VP1和清道夫受体B2(SCARB2)的表达情况。结果空白组、阳性对照组和实验组细胞的病毒空斑数分别为(52.34±12.01)、(20.88±6.47)和(18.25±6.63)个,VP1蛋白相对表达水平分别为1.87±0.14、0.77±0.11和0.74±0.09,SCARB2蛋白相对表达水平分别为0.96±0.13、0.26±0.10和0.23±0.09。实验组的上述指标与空白组比较,差异均有统计学意义(P<0.01,P<0.05)。结论川楝素能通过降低CA16中SCARB2的表达,从而抑制其在细胞内的复制。

Objective To investigate the inhibitory effect of Toosendanin from Toosendan Fructus extract on the replication of coxsackie virus A16(CA16)and its related molecular mechanism.Methods HCT116 cells were divided into blank group,positive control group and experimental group.CA16 infected cells were cultured in the experimental group with 50μg·mL^(-1) Toosendanin,in the positive control group with 24.4μg·mL^(-1) ribavirin,and in the blank group with normal culture.The effect of Toosendanin on the activity of HCT116 cells infected by Coxsackie virus A16 was detected by CCK-8 assay.Viral plaque assay was used to detect the infection and replication ability of CA16.The expression of viral structural protein VP1 and scavenger receptor B2(SCARB2)were detected by Western blotting.Results The number of viral plaque in blank group,positive control group and experimental group was 52.34±12.01,20.88±6.47 and 18.25±6.63;the expression levels of VP1 protein were 1.87±0.14,0.77±0.11 and 0.74±0.09;the expression levels of SCARB2 protein were 0.96±0.13,0.26±0.10 and 0.23±0.09,respectively.There were statistically significant differences between experimental group and blank group(all P<0.05).Conclusion Toosendanin can inhibit the replication of CA16 by decreasing the expression of SCARB2.