摘要
目的观察泡球蚴感染中期BALB/c小鼠所致肝纤维化时Ⅰ型胶原(Col1a1)、Ⅲ型胶原(Col3a1)、Ⅳ型胶原(Col4a1)基因的表达及意义。方法将雌性BALB/c小鼠随机分为实验组(8只)和对照组(5只)。实验组小鼠开腹,肉眼直视下左肝叶内注射泡球蚴混悬液,建立疾病动物模型;对照组以同样方法注射等量生理盐水。于感染中期第12周,颈椎脱臼处死小鼠,实验组取病变组织和病变邻近肝组织,对照组取注射部位所在肝组织以及临近肝组织,用4%甲醛固定,常规石蜡包埋,HE、Masson染色后观察形态学改变和肝纤维化程度。取病变临近肝组织,提取总RNA,反转录合成cDNA,实时荧光定量PCR(Real-time PCR)检测Col1a1、Col3a1、Col4a1基因的表达,计量结果用相对单位(AU)表示。结果泡球蚴感染组和对照组Collal基因表达量分别为(2.155±0.809)AU和(1.000±0.221)AU,差异有统计学意义(t=3.427,P<0.05);Col3a1基因表达量分别为(18.641±10.244)AU和(1.000±0.412)AU,差异有统计学意义(t=10.324,P<0.01);Col4a1基因表达量分别为(0.894±0.495)AU和(1.000±0.381)AU,差异无统计学意义(t=1.932,P>0.05)。Masson染色检查泡球蚴感染组肝纤维化评分为2.72±1.04,对照组为1.0,差异有统计学意义(t=4.105,P<0.05)。结论在泡球蚴感染小鼠中期,引起肝纤维化的胶原主要为Ⅲ型和Ⅰ型。
Objective To investigate the expression and significance of Col1a1,Col3a1,and Col4a1 in the liver of BALB/c mice infected with Echinococcus multilocularis in the middle stages of infection.Methods BALB/c mice were randomly divided into an experimental group and control group.The abdomen of mice in the experimental group was opened and mice were injected with 0.1 ml of an E.multilocularis metacestode suspension in the left lobe of the liver;control mice were injected with 0.1 ml saline using the same surgical procedure.Mice were sacrificed at 12 weeks.Liver samples were formalin-fixed and embedded in paraffin and prepared in 4 μm-thick sections.Pathologic changes and the degree of liver fibrosis in all samples from mice infected with E.multilocularis were observed using HE staining and Masson staining.Total RNA was extracted from tissue close to the E.multilocularis lesion using Trizol.cDNA was obtained by reverse transcriptional polymerase chain reaction.The expression of Col1a1,Col3a1,and Col4a1 was measured with real-time PCR,and Col expression was calculated in arbitrary units（AU）.Results Compared to the control group,mice infected with E.multilocularis had a higher level of expression of Col1a1 [（2.155±0.809） AU vs.（1.000±0.221） AU,P〈0.05）] and Col3a1 [（18.641±10.244） AU vs.（1.000±0.412） AU,P〈0.01）].There were no differences in the expression of Col4a1 for infected mice and the control group [（0.894±0.495） AU vs.（1.000±0.381） AU,P〉0.05）].Conclusion In the middle stages of E.multilocularis infection,liver fibrosis occurred,resulting in high levels of Col1a1 and Col3a1 expression.
出处
《中国病原生物学杂志》
CSCD
北大核心
2012年第9期653-656,共4页
Journal of Pathogen Biology
基金
新疆维吾尔自治区自然科学基金青年基金项目(No.2010211B19)
关键词
胶原
泡球蚴
肝纤维化
实时荧光定量PCR
Collagen
Echinococcus multilocularis
liver fibrosis
real time PCR
