本文關(guān)鍵詞： 心肌炎 柯薩奇病毒感染 B10細(xì)胞 調(diào)節(jié)性B細(xì)胞　出處：《廣西醫(yī)科大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：觀察柯薩奇病毒B3 (Coxsackie virus B3, CVB3)誘導(dǎo)的小鼠急性病毒性心肌炎(Acute viral myocarditis, AVMC)發(fā)病過(guò)程中不同時(shí)點(diǎn)脾臟B10細(xì)胞亞群比例的變化,初步探討B(tài)10細(xì)胞是否參與AVMC的炎癥過(guò)程。方法：利用CVB3感染無(wú)特定病原級(jí)(Specific pathogen free, SPF)雄性Balb/c小鼠建立AVMC模型。AVMC組(n=20)小鼠腹腔注射CVB3病毒液,對(duì)照組(n=10)小鼠腹腔注射等量磷酸鹽緩沖液(Phosphate buffered saline, PBS);依據(jù)注射CVB3或PBS后的時(shí)間,AVMC組和對(duì)照組小鼠均隨機(jī)分為1周和2周兩個(gè)時(shí)點(diǎn)亞組。各亞組在到達(dá)相應(yīng)觀察時(shí)點(diǎn)后處死存活小鼠,無(wú)菌留取心臟及脾臟組織標(biāo)本。取心臟組織作石蠟切片,行蘇木素伊紅(Hematoxylin-Eosin, HE)染色,觀察心肌組織病理改變并計(jì)算心肌病理積分；實(shí)時(shí)熒光定量逆轉(zhuǎn)錄聚合酶鏈反應(yīng)(real time-Polymerase Chain Reaction, RT-PCR)檢測(cè)心肌組織中CVB3 mRNA的表達(dá)水平；流式細(xì)胞術(shù)檢測(cè)小鼠脾臟CD19+IL-10+細(xì)胞(即B10細(xì)胞)亞群和CD19+CDldhiCD5+細(xì)胞亞群占CD19+細(xì)胞的比例。結(jié)果：AVMC組各亞組小鼠心肌炎癥水平明顯高于對(duì)照組相應(yīng)亞組,AVMC組中1周亞組小鼠心肌組織病理積分低于2周亞組,差異均有統(tǒng)計(jì)學(xué)意義(P均0.05)；與對(duì)照組相比,AVMC組各亞組小鼠心肌組織CVB3mRNA表達(dá)水平均明顯升高,且AVMC組中1周亞組小鼠心肌CVB3 mRNA表達(dá)水平高于2周亞組,差異均有統(tǒng)計(jì)學(xué)意義(P均0.05)。AVMC組小鼠脾臟B10細(xì)胞亞群比例和CD19+CDldhiCD5+細(xì)胞亞群比例均高于對(duì)照組,且AVMC組1周亞組小鼠脾臟B10細(xì)胞亞群比例和CD19+CDldhiCD5+細(xì)胞亞群比例均高于2周亞組,差異均有統(tǒng)計(jì)學(xué)意義(P均0.05)。AVMC小鼠發(fā)病過(guò)程中,脾臟B10細(xì)胞亞群比例與CD19+CDldhiCD5+細(xì)胞亞群比例及心肌組織中CVB3mRNA表達(dá)三者相平行,而與心肌組織炎癥水平相反。結(jié)論：B10細(xì)胞在急性病毒性心肌炎小鼠脾臟中所占比例增高,其參與了急性病毒性心肌炎的炎癥過(guò)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.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R542.21

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