摘要
目的炎性因子的分泌水平与严重发热伴血小板减少综合征(SFTS)的严重程度密切相关。文章主要探讨严重发热伴血小板减少综合征病毒(SFTSV)感染小鼠巨噬细胞对Toll样受体2(TLR2)和炎性因子表达的影响。方法将SFTSV感染小鼠巨噬细胞,采用qPCR、Western blot和ELISA方法检测小鼠巨噬细胞后0、6、12、24、36、48、72 h的TLR2、肿瘤坏死因子-α(TNF-α)、干扰素-β(IFN-β)、白细胞介素-6(IL-6)、IL-10等炎性因子的表达变化。结果qPCR结果显示,0、6、12、24、36、48、72 h TNF-α的表达量分别为0.85±0.14、15.23±2.45、28.67±1.59、94.52±7.05、55.86±1.82、23.55±6.15、9.76±2.03,IFN-βmRNA分别为0.93±0.21、1.55±0.33、14.51±1.98、55.16±3.64、26.57±1.49、9.22±0.51、5.18±1.06,以上结果提示0~72 h感染的小鼠腹腔巨噬细胞生成的TNF-α和IFN-βmRNA水平呈现先上升(24h最高)后下降的趋势,与0 h比较,6、12、24、36、48、72 hTNF-α和IFN-βmRNA水平差异均有统计学意义(P<0.05);而IL-6和IL-10 mRNA则呈现持续升高,与0 h比较,6、12、24、36、48、72 h差异均有统计学意义(P<0.05)。ELISA结果显示TNF-α、IFN-β、IL-6、IL-10蛋白的表达均呈逐渐升高的趋势,TNF-α的0、72 h分别为(38.31±4.25、140.41±23.45)pg/mL,IFN-β蛋白0、72 h分别为(17.56±0.66、1084.93±111.42)pg/mL,IL-6蛋白0、72 h分别为(113.30±0.07、2302.32±134.09)pg/mL,IL-10蛋白0、72 h分别为(515.00±21.21、2590.40±226.19)pg/mL,以上结果提示TNF-α、IFN-β、IL-6、IL-10蛋白24、36、48、72 h均较0 h明显升高(P<0.05)。小鼠腹腔巨噬细胞生成的TLR2 mRNA表达均呈现先上升后下降的趋势,在24 h上升达到最高水平,每个时间点与0 h相比差异均有统计学意义(P<0.01)。TLR2蛋白在感染后随着时间延长表达逐渐升高。结论SFTSV感染能促进巨噬细胞上调TLR2的表达,进而可能导致炎性因子分泌增加。
Objective The secretion level of inflammatory factors is closely related to the severity of severe fever with thrombocytopenia syndrome(SFTS).This paper mainly discussed the effect of SFTSV infection on the expression of Toll-like receptor 2(TLR2)and inflammatory factors in macrophages in mice.MethodsThe expression of TLR2 and TNF-α,IFN-β,IL-6,IL-10 and other inflammatory factors were observed at 0,6,12,24,36,48,72 hours after SFTSV infected mice macrophages by q PCR,Western blot and ELISA.ResultsQPCR results showed that the m RNA levels of TNF-αat 0,6,12,24,36,48,72 h were 0.85±0.14,15.23±2.45,28.67±1.59,94.52±7.05,55.86±1.82,23.55±6.15,9.76±2.03,and the m RNA levels of IFN-βwere 0.93±0.21,1.55±0.33,14.51±1.98,55.16±3.64,26.57±1.49,9.22±0.51,5.18±1.06.QPCR results showed that the m RNA levels of TNF-αand IFN-βproduced by abdominal macrophages in the infected mice showed a trend of first increasing(the highest at the 24 hour)and then decreasing,and the difference of TNF-αand IFN-βm RNA levels at other time points was statistically significant compared with that at the 0 hour(P<0.05).However,IL-6 and IL-10 m RNA levels continued to increase,and the difference at other time points was statistically significant(P<0.05).ELISA results showed that the expression of four inflammatory factors showed a trend of gradual increase:TNF-α0 and 72 h were(38.31±4.25,140.41±23.45)pg/m L;IFN-β0 th and 72 th were(17.56±0.66,1084.93±111.42)pg/m L;IL-6 protein 0 and 72 h were(113.30±0.07,2302.32±134.09)pg/m L;IL-10 protein 0 and 72 h were(515.00±21.21,2590.40±226.19)pg/m L,respectively,with significant increases at the 24,36,48 and 72 hours compared with that at the 0 hour(P<0.05).The expression of TLR2 m RNA generated by mouse peritoneal macrophages showed a trend of first increasing and then decreasing,and increased to the highest level at 24 h,and the difference between each time point and 0 h was statistically significant(P<0.01).The expression of TLR2 gradually increased after infection with time
作者
颜莹莹
朱旭辉
胡南南
刘源
朱进
金柯
韩亚萍
李军
YAN Ying-ying;ZHU Xu-hui;HU Nan-nan;LIU Yuan;ZHU Jin;JIN Ke;HAN Ya-ping;LI Jun(Department of Infectious Diseases,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,Jiangsu,China;Center for Disease Control and Prevention in Eastern Theater,Nanjing 210002,Jiangsu,China)
出处
《医学研究生学报》
CAS
北大核心
2019年第11期1164-1168,共5页
Journal of Medical Postgraduates
基金
国家科技重大专项课题(2016ZX10002005-001-009)
