复方苦芩总生物碱保护F81细胞免受犬细小病毒损伤作用的探究

Study on the Effect of Total Alkaloids in Kuqin Compound of Protecting F81 Cells Against Canine Parvovirus

为了探讨中药复方苦芩总生物碱(Total Alkaloids in Kuqin Compound,TAKC)保护F81细胞免受犬细小病毒(Canine Parvovirus,CPV)损伤的作用机理,用CPV感染F81细胞,选择不同浓度的TAKC进行体外抗CPV感染F81细胞试验,采用琥珀酸脱氢酶活力测定法检测TAKC对F81细胞的保护率,用RT-PCR法检测TAKC处理对感染CPV的F81细胞中caspase-3基因mRNA表达水平的影响.结果显示,TAKC对F81细胞有保护作用,其在整个病毒复制过程中(直接灭活作用、阻滞作用、吸附抑制作用、穿入抑制作用和复制抑制作用)均有明显的影响,其中阻滞作用、穿入抑制作用和复制抑制作用效果最明显,对细胞的保护率分别能达到31.19%,20.54%,32.28%.同时使感染CPV的F81细胞的caspase-3基因表达水平显著降低(p<0.01),表达水平与模型组的比值为1∶1.818.因此,推测TAKC可通过阻滞抑制、穿入抑制和DNA复制抑制CPV感染F81细胞和降低F81细胞中caspase-3基因表达水平,起到保护F81细胞免受CPV损伤的作用.

An experiment was conducted to investigate the possible mechanism of total alkaloids in compound kuqin（TACK）protecting F81cells（feline kidney cells）against canine parvovirus（CPV）.In in vitro tests,F81 cells were infected with CPV and treated with TACK at different concentrations.The MTT method was used to detect the protection rate of F81 cells,and RT-PCR was used to test the expression of caspase-3mRNA.The results showed that TACK had protective effects on F81 cells throughout the process of virus replication,including direct inactivation effect,blocking effect,adsorption inhibition effect,penetration inhibition effect and replication inhibition effect.The protection of the effects of blocking inhibition,penetration inhibition and replication inhibition was 31.19%,20.54% and 32.28%,respectively.TACK significantly decreased the expression level of caspase-3mRNA in F81 cells infected with CPV,the ratio being 1∶1.818,as compared with the model group.It is,therefore,speculated that TACK can protect F81 cells from damage induced by CPV through the blocking inhibition,penetration inhibition and DNA replication inhibition effects of F81 cells against CPV infection and decrease the expression level of caspase-3mRNA.