含有CpG基序的寡聚脱氧核苷酸2395对鼠巨细胞病毒肝炎新生小鼠的治疗作用  

Therapeutic effect of neonatal mouse Cytomegalovirus hepatitis by CpG Oligodeoxynucleotide 2395

作　　者：潘兆军[1] 周利[2] 王军[3] 韩良荣[1] 曹杨[3] 万雪媛[3] 孟令建[3] 

机构地区：[1]扬州大学医学院附属淮安市妇幼保健院新生儿科,江苏淮安223002 [2]徐州医学院附属淮安医院儿科 [3]徐州医学院附属医院新生儿科

出　　处：《中华实用儿科临床杂志》2014年第2期104-109,共6页Chinese Journal of Applied Clinical Pediatrics

摘　　要：目的 研究含有CpG基序的寡聚脱氧核苷酸2395（CpG ODN2395）对鼠巨细胞病毒（MCMV）肝炎新生小鼠的治疗作用及相关机制.方法 SPF级BALB/c新生小鼠,按窝随机分为对照组、病毒组和治疗组.对照组：腹腔注射9 g/L盐水20×10-6L隔日1次;病毒组：腹腔注射MCMV（病毒致半数细胞感染量TCID50=108.31/L）20×10-6L1次,从接种MCMV当日起隔日腹腔注射9 g/L盐水20×10-6L;CpG治疗组：腹腔注射MCMV（TCID50=108.31/L） 20×10-6L1次,从接种MCMV当日起隔日腹腔注射CpG ODN2395 20 mg/kg.观测小鼠的生长发育情况.HE染色观察肝脏病理,酶联免疫吸附试验检测小鼠血清ALT、IFN-α,PCR法检测肝脏MCMV DNA,反转录（RT）-PCR法检测肝脏IFN-α mRNA,Wester blot法检测肝脏的Toll样受体9（TLR9）和衔接蛋白-髓样分化标志物88 （MyD88）的表达水平.应用SPSS 16.0软件进行分析.结果 1.病毒组小鼠生长发育落后,体质量低于对照组和治疗组,7d、14 d时差异有统计学意义（F=18.919、25.543,P均＜0.05）;治疗组发育介于对照组和病毒组之间,体质量均低于对照组,在7d时差异有统计学意义（t=3.187,P ＜0.05）.2.病毒组小鼠血清ALT高于对照组和治疗组,治疗组血清ALT高于对照组,差异均有统计学意义（F=11.407、11.791、154.565,P均＜0.05）.3.肝组织病理改变：病毒组肝组织病变活动性积分（HAI）值在第3天达高峰,以后逐渐下降;治疗组肝组织HAI均值也在第3天达高峰,但肝损害明显轻于病毒组,以后肝病变逐渐减轻,在第7天和第14天时HAI均值都显著低于病毒组.4.病毒组和治疗组小鼠3d、7d和14 d时肝脏MCMV DNAPCR检测阳性,3个时间点治疗组MCMV DNA负荷量均低于病毒组.5.病毒组和治疗组的3d时血清IFN-α达高峰,7d、14 d时渐下降;治疗组的血清IFN-α高于病毒组和对照组.6.病毒组和治疗组肝脏IFN-α mRNA表达水平在3d时增加,7d时达高峰,14 d时下降,3个时间点表达水平差异有统计Objective To study the treatment effects of newborn mice murine Cytomegalovirus （MCMV） hepatitis by using CpG Oligodeoxynucleotide 2395 （CpG ODN2395）.Methods Specific pathogen-free BALB/c newborn mice were divided into 3 groups according to the broods randomly：control group,virus group and treatment group.In control group 9 g/L sodium chloride 20 × 10-6 L was intraperitoneally injected every other day.In virus group MCMV （TCID50 =108.31/L） 20 × 10-6 L was intraperitoneally injected once and 9 g/L sodium chloride solution 20 × 10-6 L was intraperitoneally injected every other day.In treatment group murine MCMV（TCID50 =10S31/L） 20 × 10-6 L was intraperitoneally injected once and from the first day CpG ODN2395 20 mg/kg was intraperitoneally injected every other day.Growths and development of mice were observed.Murine body weights were measured.Pathology of livers was observed by means of hematoxylin and eosin stain.The levels of serum ALT and IFN-α were measured by adopting enzyme linked immunosorbent assay.MCMV loads in liver were measured by way of polymerase chain reaction.The expression levels of IFN-α mRNA in liver were detected by using reverse transcription polymerase chain reaction.The expression levels of Toll-like receptor 9 （TLR9） and myeloid cell differentiation factor 88 （MyD88） in liver were detected by adopting Western blot.The results were analyzed by using SPSS 16.0 statistics software.Results 1.Compared with control group and treatment group,growth and development of virus group mice fell behind and on day 7,14 body weights were lowest（F =18.919,25.543,all P ＜ 0.05）.Growth and development of treatment group mice were between control group and virus group.Body weights of treatment group mice were lower than those of control group,and there was statistical difference on day 7 （t =3.187,P ＜ 0.05）.2.Compared with control group and treatment group,the levels of ALT in virus group was highest.ALT levels of treatment group were higher than those of control group

关 键 词：巨细胞病毒 含有CpG基序的寡聚脱氧核苷酸2395 肝炎 小鼠 

分 类 号：R722.1[医药卫生—儿科]