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目的探讨建立鼠巨细胞病毒(MCMV)感染致新生小鼠肝炎模型的可行性。方法48只日龄24 h内新生BALB/c小鼠随机分为实验组和对照组,实验组每只小鼠腹腔注射MCMV Smith株20μl TCID50(104.31/0.1 ml),对照组注射无菌生理盐水20μl,注射第3、7、14天取血清和肝脏,检测血清ALT水平,肝脏组织HE染色后用光镜检查组织病理损害,同时提取肝脏组织的DNA,应用MCMV及β-actin引物行PCR扩增,然后跑电泳和测序。结果实验组小鼠ALT水平(U/L)在3天时即明显升高,7天达高峰,14天时有所下降,与对照组比较,差异均有统计学意义[3天:(58.7±11.5)比(25.0±10.6),7天:(169.6±57.4)比(25.1±8.4),14天:(157.3±15.5)比(26.5±9.4),P均<0.01]。实验组小鼠肝组织内均可见肝细胞气球样变性,点灶状坏死,门管区炎性细胞浸润,肝细胞核内病毒包涵体,7天时最严重;对照组肝细胞无相似病理变化。实验组MCMV-DNA PCR电泳全部出现阳性条带,阳性条带测序结果与MCMV基因序列的同源性完全相符。结论 MCMV能侵袭BALB/c新生小鼠引起肝炎,这种模拟人类巨细胞病毒肝炎的新生小鼠模型的建立为该病动物实验研究提供了可能。
Objective To investigate the feasibility of establishing a mouse model of neonatal hepatitis by cytomegalovirus (MCMV) infection. Methods 48 neonatal BALB / c mice were randomly divided into experimental group and control group within 24 h. In the experimental group, 20 μl TCID50 (104.31 / 0.1 ml) of MCMV Smith strain was injected intraperitoneally into the experimental group. The control group was injected with sterile saline Serum and liver were collected on the 3rd, 7th and 14th day after injection. Serum ALT levels were measured. The liver tissues were stained with HE for histopathological damage by light microscopy. DNA from liver tissues was also extracted. MCMV and β-actin primers were used for PCR amplification Increase then run for electrophoresis and sequencing. Results The ALT level (U / L) in experimental group increased significantly at 3 days, peaked at 7 days, decreased at 14 days, and the difference was statistically significant compared with control group [3 days: (58.7 ± 11.5 ) (25.0 ± 10.6), 7 days: (169.6 ± 57.4) vs (25.1 ± 8.4), 14 days: (157.3 ± 15.5) vs. (26.5 ± 9.4), P <0.01; In experimental group, hepatic cell balloon-like degeneration, focal necrosis, inflammatory cell infiltration in portal area, and virus inclusion body in hepatocyte were all observed in the liver tissue of mice in experiment group. The hepatic cells in control group had no similar pathological changes. The MCMV-DNA PCR electrophoresis in the experimental group all showed positive bands, and the positive band sequencing results were completely consistent with the homology of the MCMV gene sequences. Conclusions MCMV can infect neonatal mice with BALB / c to cause hepatitis. The establishment of a neonatal mouse model of human cytomegalovirus hepatitis may provide experimental evidence for animal studies.