B10细胞在急性病毒性心肌炎小鼠中的变化

发布时间：2018-02-15 12:03

本文关键词： 心肌炎柯萨奇病毒感染 B10细胞调节性B细胞　出处：《广西医科大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的：观察柯萨奇病毒B3 (Coxsackie virus B3, CVB3)诱导的小鼠急性病毒性心肌炎(Acute viral myocarditis, AVMC)发病过程中不同时点脾脏B10细胞亚群比例的变化,初步探讨B10细胞是否参与AVMC的炎症过程。方法：利用CVB3感染无特定病原级(Specific pathogen free, SPF)雄性Balb/c小鼠建立AVMC模型。AVMC组(n=20)小鼠腹腔注射CVB3病毒液,对照组(n=10)小鼠腹腔注射等量磷酸盐缓冲液(Phosphate buffered saline, PBS);依据注射CVB3或PBS后的时间,AVMC组和对照组小鼠均随机分为1周和2周两个时点亚组。各亚组在到达相应观察时点后处死存活小鼠,无菌留取心脏及脾脏组织标本。取心脏组织作石蜡切片,行苏木素伊红(Hematoxylin-Eosin, HE)染色,观察心肌组织病理改变并计算心肌病理积分；实时荧光定量逆转录聚合酶链反应(real time-Polymerase Chain Reaction, RT-PCR)检测心肌组织中CVB3 mRNA的表达水平；流式细胞术检测小鼠脾脏CD19+IL-10+细胞(即B10细胞)亚群和CD19+CDldhiCD5+细胞亚群占CD19+细胞的比例。结果：AVMC组各亚组小鼠心肌炎症水平明显高于对照组相应亚组,AVMC组中1周亚组小鼠心肌组织病理积分低于2周亚组,差异均有统计学意义(P均0.05)；与对照组相比,AVMC组各亚组小鼠心肌组织CVB3mRNA表达水平均明显升高,且AVMC组中1周亚组小鼠心肌CVB3 mRNA表达水平高于2周亚组,差异均有统计学意义(P均0.05)。AVMC组小鼠脾脏B10细胞亚群比例和CD19+CDldhiCD5+细胞亚群比例均高于对照组,且AVMC组1周亚组小鼠脾脏B10细胞亚群比例和CD19+CDldhiCD5+细胞亚群比例均高于2周亚组,差异均有统计学意义(P均0.05)。AVMC小鼠发病过程中,脾脏B10细胞亚群比例与CD19+CDldhiCD5+细胞亚群比例及心肌组织中CVB3mRNA表达三者相平行,而与心肌组织炎症水平相反。结论：B10细胞在急性病毒性心肌炎小鼠脾脏中所占比例增高,其参与了急性病毒性心肌炎的炎症过程。
[Abstract]:Objective: to observe the changes of spleen B10 cell subsets at different time points during acute viral myocarditis (AVMC) induced by Coxsackie virus B3 (CVB3). Methods: a AVMC model was established in male Balb/c mice infected with CVB3 without specific pathogen, and the AVMC model was established by intraperitoneal injection of CVB3 virus. Mice in the control group were intraperitoneally injected with Phosphate buffered Saline (PBSN). According to the time after injection of CVB3 or PBS, the control group and the control group were randomly divided into two subgroups: one week and two weeks. Each subgroup reached the corresponding observation time point. And then killed the surviving mice, The heart tissue was taken as paraffin section and stained with Hematoxylin-Eosin (HEH). The pathological changes of myocardial tissue were observed and the pathological score of cardiomyopathy was calculated. Real time-Polymerase Chain reaction (RT-PCR) was used to detect the expression of CVB3 mRNA in myocardium. The subsets of CD19 IL-10 cells (B10 cells) and CD19 CDldhiCD5 cells in the spleen of mice were detected by flow cytometry. Results the myocardial inflammation level of mice in the subgroup of CD19 CDldhiCD5 was significantly higher than that in the control group. The pathological score of myocardial tissue in group B was lower than that in subgroup 2 weeks. Compared with the control group, the expression of CVB3mRNA in the myocardium of each subgroup was significantly higher than that of the control group, and the level of myocardial CVB3mRNA expression in 1 week subgroup was higher than that in 2 week subgroup. The percentage of B10 cell subsets and CD19 CDldhiCD5 subsets in spleen of AVMC group was higher than that of control group, and the percentage of B10 cell subgroup and CD19 CDldhiCD5 subgroup in AVMC group was higher than that in control group. The difference was statistically significant (P < 0.05). The proportion of B10 cell subsets in spleen was parallel to that of CD19 CDldhiCD5 cell subsets and CVB3mRNA expression in myocardium. Conclusion the proportion of B10 cells in the spleen of mice with acute viral myocarditis is increased, and it is involved in the inflammatory process of acute viral myocarditis.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R542.21

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