摘要
目的通过建立新生小鼠巨细胞病毒(CMV)肝炎模型,检测黑色素瘤缺乏因子2 (AIM2)、半胱氨酸天冬氨酸蛋白酶-1 (Caspase-1)的表达水平,探讨其在CMV肝炎中的作用及意义,为寻找治疗CMV肝炎新靶点提供理论依据。方法48只新生BALB/c小鼠随机分为观察组及对照组,各24只,每组又按照3 d、7 d、14 d分为三个亚组,每组各8只。观察组于生后12 h内腹腔注射组织半数细胞感染量(TCID50)为104. 44U/ml的CMV悬液20μl,建立CMV肝炎模型,对照组腹腔注射等容积无菌生理盐水,于造模后3 d、7 d、14 d留取静脉血及肝组织,HE染色观察肝脏病理变化,ELISA法检测血清谷丙转氨酶(ALT)表达水平,PCR检测肝组织CMV-DNA表达,Western blot检测肝组织AIM2、Caspase-1表达水平及变化。结果CMV感染后,相较对照组,观察组小鼠出现皮毛稀疏、纳差、少动、反应差、生长发育落后等表现。观察组小鼠肝组织HE染色可见明显病理损害,对照组未见明显异常。观察组各时间点小鼠肝脏组织CMV-DNA PCR电泳均见阳性条带,对照组无相应条带。观察组小鼠肝脏组织中AIM2、Caspase-1表达明显高于对照组,且各时间点间差异有统计学意义(均P<0. 05);观察组表达于3 d时可见升高,7 d时达峰值,14 d时较前降低,差异有统计学意义(均P<0. 05)。观察组小鼠血清ALT水平明显高于对照组,且各时间点间差异有统计学意义(均P<0. 05),观察组3 d时可见升高,7 d时达最高,14 d时较前下降,差异有统计学意义(均P<0. 05)。观察组小鼠肝脏组织中AIM2、Caspase-1蛋白表达量与血清ALT呈正相关性(均P<0. 05)。结论AIM2炎症体介导的细胞焦亡程序可能为CMV肝炎的致病机制之一,且肝脏组织中AIM2、Caspase-1表达越高,肝细胞损伤越重。
Objective To investigate the expressions and significances of AIM2, caspase-1 in cytomegalovirus-induced hepatitis of neonatal mice. Methods 48 BALB/c mice were randomly divided into blank group （group C） and virus group （group A）, 24 mice in each group. Mice in group A were received intraperitoneal injection of MCMV suspension within 24 hours after birth at the dosage of 20μll and tissue culture infective dose of 104.44 U/ml, meanwhile, the same volume of normal saline was injected into mice in group C. The patholog- ical changes of liver were observed by HE staining. MCMV-DNA was detected by PCR, and the protein expressions of AIM2, caspase-1 were analyzed by Western blot. The level of ALT was detected by ELISA. Results After MCMV infection, mice appeared sparse fur, poor appetite, less activity, unresponsiveness to stimuli, growth retardation, etc. The pathological changes were found in liver of virus group. MCMV-DNA was expressed in virus group, but absent in control group. The protein expressions of AIM2, caspase-1 were increased in virus group than those in contron group. The protein expressions of AIM2, caspase- 1 in virus group were increased at day3, peaked at day7 and decreased at dayl4 （ P〈0. 05 ） . In virus group, the expression of ALT was higher than that in control group at the same time point （P〈0. 05 ） . In virus group, the expressions of AIM2, caspase- 1 were positively correlated with ALT at each time point. Conclusion The apoptosis induced by AIM2 inflammasome might be the pathogenesis in hepatopathy caused by MCMV, the higher the expressions of AIM2, caspase-1 in liver tissue, the more serious the liver cell injury.
作者
汪浩森
王军
邱文尧
苏飚
WANG Hao-Sen;WANG Jun;QIU Wen-Yao(Xuzhou Children＇s Hospital,Xuzhou,Jiangsu 221000,China)
出处
《中国妇幼保健》
CAS
2018年第21期4971-4975,共5页
Maternal and Child Health Care of China
