靛玉红对ORFV感染淋巴细胞的增殖、细胞因子含量及CD分子表达量的影响  

Effects of indirubin on proliferation,cytokine content and expression of CD molecules in ORFV-infected lymphocytes

作　　者：曾琴 富国文 赵晏羚 张朝龙 吴姣 樊月圆 四朗玉珍[3] 舒相华 宋春莲 ZENG Qin;FU Guowen;ZHAO Yanling;ZHANG Chaolong;WU Jiao;FAN Yueyuan;Silangyuzhen;SHU Xianghua;SONG Chunlian(College of Veterinary Medicine,Yunnan Agricultural University,Kunming 650201,China;Dali City Center for Animal Disease control and Prevention,Dali 671003,China;Institute of Animal Husbandry and Veterinary Medicine,Tibet Autonomous Region Academy of Agriculture and Animal Husbandry Sciences,Lhasa 850009,China)

机构地区：[1]云南农业大学动物医学院,昆明650201 [2]大理市动物疫病预防控制中心,云南大理671003 [3]西藏自治区农牧科学院畜牧兽医研究所,拉萨850009

出　　处：《黑龙江畜牧兽医》2023年第4期104-109,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金：西藏自治区“十四五”重大科技专项(XZ202101ZD0001N);云南省农业重大科技专项计划项目(202102AE090007);云南省教育厅科学研究基金项目(2020J0257)。

摘　　要：为了研究靛玉红对羊传染性脓疱病毒(Ovine contagious pustular dermatitis virus,ORFV)感染淋巴细胞的增殖、细胞因子质量浓度及CD分子表达量的影响,在靛玉红对ORFV感染淋巴细胞增殖的影响试验中,分为对照组、ORFV感染组、靛玉红(0.2μg/mL)+ORFV组、靛玉红(0.2μg/mL)组,采用CCK-8法检测靛玉红对ORFV感染淋巴细胞增殖的影响。另外,在靛玉红对ORFV感染细胞上清液中细胞因子质量浓度及CD分子表达量的影响试验中,分为高浓度靛玉红(0.2μg/mL)+ORFV组、中浓度靛玉红(0.1μg/mL)+ORFV组、低浓度靛玉红(0.05μg/mL)+ORFV组,于96孔培养板中,每孔加入淋巴细胞悬液100μL,之后加ORFV病毒悬液100μL,吸附2 h,弃病毒液,分别加入终浓度为0.2,0.1,0.05μg/mL的靛玉红溶液100μL,作用0,2,4,8,12 h,另设ORFV感染组(只加ORFV病毒悬液和RDMI-1640完全培养基)、对照组(只加RPMI-1640完全培养基),采用ELISA方法检测细胞上清液中IFN-γ、TNF-β、IL-10的质量浓度及CD4、CD8的表达量,并计算CD4/CD8。结果表明:与ORFV感染组相比,靛玉红+ORFV组淋巴细胞增殖数极显著升高(P<0.01)。与ORFV感染组比较,作用4,8,12 h后,高、中、低浓度靛玉红+ORFV组的CD4分子表达量和CD4/CD8显著或极显著升高(P<0.05或P<0.01),同时Th1型细胞因子IFN-γ、TNF-β质量浓度及Th2型细胞因子IL-10的质量浓度不同程度升高,而CD8分子表达量降低。说明靛玉红能够增强ORFV感染淋巴细胞的增殖,促进IFN-γ、TNF-β和IL-10的分泌,提示靛玉红可能是通过调节CD4、CD8的表达来促进细胞因子的分泌,增加ORFV感染淋巴细胞的增殖,进而调节机体的免疫机能。In order to study the effects of indirubin on proliferation,cytokine content and expression of CD molecules in ORFV(Contagious pustular dermatitis virus)-infected lymphocytes,in the test of the effects of indirubin on the proliferation of ORFV-infected lymphocytes,they were divided into control group,ORFV-infected group,indirubin(0.2μg/mL)+ORFV group,and indirubin(0.2μg/mL)group.The effects of indirubin on the proliferation of ORFV-infected lymphocytes was detected by CCK-8 method.In addition,in the test of the effects of indirubin on the cytokine content and CD molecule expression in the supernatant of ORFV-infected cells,they were divided into high concentration of indirubin(0.2μg/mL)+ORFV group,medium concentration of indirubin(0.1μg/mL)+ORFV group,low concentration of indirubin(0.05μg/mL)+ORFV group.96-well plates were cultured with 100μL lymphocyte suspension in each well.After that,ORFV virus suspension was added 100μL and adsorbed for 2 h.The disease venom was discarded and indirubin solution with final concentrations of 0.2,0.1,0.05,0μg/mL was added 100μL.In addition,ORFV-infected group(only adding ORFV virus suspension and RPMI-1640 complete medium)and control group(only adding RPMI-1640 complete medium)were set up.After 0,2,4,8,12 h of the experiment,the mass concentrations of IFN-γ,TNF-βand IL-10 and the expressions of CD4 and CD8 in the supernatant were detected by ELISA,and CD4/CD8 were calculated.The results showed that the lymphocyte proliferation number was highly significantly enhanced in the indirubin+ORFV group compared with the ORFV-infected group(P<0.01).Compared with ORFV-infection group,after 4,8,12 h of the experiment,high,medium and low concentration of indirubin+ORFV group,CD4 molecular expression and CD4/CD8 were increased(P<0.05 or P<0.01),while the mass concentration of Th1 type cytokines IFN-γ,TNF-βand Th2 type cytokines IL-10 were increased,and CD8 molecule expression decreased.The results suggested that indirubin enhanced the proliferation of ORFV-infected lymphocytes

关 键 词：羊传染性脓疱病毒 靛玉红 淋巴细胞 细胞因子 酶联免疫吸附试验 免疫调节 

分 类 号：S853.7[农业科学—临床兽医学]