摘要
为了探讨H9亚型禽流感病毒对MDCK细胞增殖及其培养基过氧化氢含量的影响,将不同剂量的H9亚型禽流感病毒接种MDCK细胞单层,吸附1.5h后弃去再加入新的DMEM维持液,每隔12h取上清液测定HA滴度、乳酸脱氢酶(LDH)活力以及过氧化氢(H2O2)含量。同时,本试验采用MTT法测定不同剂量病毒接种对MDCK细胞增殖的影响。结果表明,以10-3EID50、10-4EID50病毒剂量接种细胞后,病毒HA滴度分别在36h和48h达到滴度最大值(9log2)以及LHD活力的最大值,上清液中H2O2含量在48h时显著(P<0.05)高于空白对照组和低剂量病毒接种组;接种病毒后36h以内对MDCK细胞生长具有显著地(P<0.05)促进作用,之后会显著(P<0.05)抑制细胞的增殖。综上所述,H9亚型禽流感病毒能在前期促进MDCK细胞的增殖,随着病毒增殖对细胞损伤加大,LDH漏出量增加,促凋亡因子H2O2参与介导了这一过程。
This study was aimed to determine the effects of subtype H9 avian influenza virus cultured with MDCK cells on cell proliferation and content of hydrogen peroxide in medium. The HA titer,lactate dehydrogenase （LDH） activity and hydrogen peroxide （H2O2）content in the supernatant were determined every 12 hours from subtype H9 avian influenza virus cultured with monolayer MDCK cells. In this study, MTT assay was used for investigating the effects of virus inoculation at different doses on the proliferation of MDCK ceils. The results showed that at the dose of 10-3 EID50 or 10-4 EID50,tbe virus＇ HA titer reached the maximum （9log2）and the maximun of LHD activity at 36h or 48 h,respectively. In addition, H2O2 content in the superuatant increased significantly （P〈0.05）, as compared with the control group or group treated with low dose of virus. After cultured with subtype H9 avian influenza virus,the proliteration of MDCK cells was promoted significantly （P〈0.05）before 36 h,and then inhibited significantly （P〈0.05）,compared with the control group. In summary,subtype H9 avian influenza virus could promote the proliferation of MDCK cells. With the increasing of cell damage aroused by virus proliferation,LDH leakage increased,which was probably mediated by apoptotic factor of H2O2 in this process.
出处
《中国家禽》
北大核心
2012年第17期11-15,共5页
China Poultry
基金
国家自然科学基金项目(30871900)
