摘要
采用CCK8法测定小槐花提取物在MDCK细胞上的最大安全浓度(Maximum no-cytotoxic concentration,MNTC);建立甲型流感病毒A/PR8/34(H1N1,PR8)感染MDCK细胞模型,CCK8法检测其对病毒感染细胞病变的抑制作用;利用荧光定量PCR及免疫印迹方法,检测小槐花提取物对流感病毒基因、蛋白,以及宿主炎症相关转录因子和炎性细胞因子表达的影响。细胞毒性试验结果显示,小槐花提取物对MDCK细胞的MNTC为100 mg/L;qRT-PCR结果显示,小槐花提取物能显著抑制PR8流感病毒M与NP基因的mRNA表达(P<0.01),也可显著下调病毒感染细胞的转录因子NF-κB p65、STAT3及炎性细胞因子IL-1β、TNF-α基因的表达水平,且其抑制作用与药物浓度正相关;免疫印迹法检测显示,100 mg/L的小槐花提取物极显著抑制了流感病毒M1蛋白的表达(P<0.01)。小槐花提取物具有较显著的体外抗流感病毒效果,此研究结果为从小槐花中鉴定具有抗流感病毒的确切药效成分奠定基础。
The maximum safe concentration(MNTC)of Desmodium caudatum extracts(DCE)on MDCK cells was determined by CCK8 method.The model of MDCK cells infected with influenza virus A/PR8/34(H1N1,PR8)was established,and its inhibitory effect on cytopathic changes caused by viral infection was detected using CCK8 method.Using fluorescence quantitative PCR method,the effects of DCE on influenza virus genes,proteins and expression of host inflammatory transcription factors and inflammatory cytokines were detected.The cytotoxicity test results showed that the MNTC of DCE on MDCK cells was 100 mg/L.The qRT-PCR results showed that DCE could significantly reduce mRNA expression of M and NP genes of PR8 influenza virus(P<0.01),and also significantly downregulated the expression levels of inflammatory transcription factors NF-kB p65 and STAT3,and inflammatory cytokines IL-1βand TNF-α,whose inhibitory effect was positively correlated with drug concentration.Immunoblotting assay showed that DCE of high concentration(100 mg/L)significantly inhibited the expression of influenza virus M1 protein(P<0.01).These results indicate that Desmodium caudatum extracts have significant in vitro anti-influenza virus effects,and lays foundation for identifying the exact pharmacological components with anti influenza virus effects from Desmodium caudatum.
作者
徐国双
金新
郭艺迪
张茂林
段铭
关振宏
XU Guoshuang;JIN Xin;GUO Yidi;ZHANG Maolin;DUAN Ming;GUAN Zhenhong(Key Laboratory for Zoonosis Research,Ministry of Education,Institute of Zoonosis,Jilin University,Changchun 130062,China)
出处
《吉林农业大学学报》
CAS
CSCD
北大核心
2024年第1期148-154,共7页
Journal of Jilin Agricultural University
基金
国家自然科学基金项目(32072902,31970154)。
