肺炎衣原体感染影响胰岛β细胞增殖和功能的体外研究  

Study on the infection of chlamydia pneumoniae affecting the proliferation and function of pancreatic βcells in vitro

作　　者：陈思思[1] 于德民[1] 周赛君[1] 李君文[2] 王彤[1] 宋苗瑞[1] 张睿[1] 于珮[1] 

机构地区：[1]大津医科大学代谢病医院肾痫血液透析科卫生部激素与发育重点实验室,300070 [2]中国人民解放军军事医学科学院卫生学环境医学研究所,天津300050

出　　处：《国际内分泌代谢杂志》2013年第3期150-154,F0003,共6页International Journal of Endocrinology and Metabolism

基　　金：国家自然科学基金资助项目（30971393,81070645）：天津市自然科学基金资助项目（10JCYBJCl2000）;天津市卫生局科技基金重点项日及一般项日（09KZ01,09KZ89）;天津医科大学科技基金资助项日（2009ky25）

摘　　要：目的探讨肺炎衣原体（C．pn）感染对胰岛β细胞增殖和功能的影响。方法将C．pn在人喉癌细胞系（HEp－2细胞）中增殖培养后，以感染复数为0．08、0．20，干预时间为6，24h的c．pn感染体外培养的胰岛β细胞系（INS－1细胞），通过光镜下观察细胞形态变化及免疫荧光单克隆抗体染色方法确认感染模型的建立。在不同感染复数（0，0．04，0．08，0．10，0．12，0．1d，0．16，0．20）、不同干预时间（2，4，6，8，16，24，36h）的C．pn作用下，利用四甲基偶氮唑蓝法、台盼蓝活细胞计数及流式细胞仪观察C．pn感染对INS—l细胞增殖和凋亡的影响；并通过放射免疫法测定c．pn感染后经高糖（25mmol／L）和低糖（5．6mmol／L）刺激的INS－1细胞胰岛素分泌量。结果与正常对照组相比，小剂量（0．08）加短时间（6h）C．pn感染组INS－1细胞密度增加，细胞间隙变窄，大剂量（0．20）加长时间（24h）C．pn感染组INS—I细胞骤缩、变圆、仍聚团生长。免疫荧光单克隆抗体染色显示特异性亮绿色荧光，证实C．pn感染INS—l细胞的体外模型成功建立。与正常对照组相比，小剂量（〈0．20）加短时间（〈24h）的C．pn感染促进INS-1细胞增殖（t=-8．907，P〈0．05）及INS-1细胞分泌胰岛素（t=-9．186，P〈0．05），而大剂量（〉0．20）加长时问（〉24h）的C．pn感染则促进INS-1细胞凋亡（t=-37．306，P〈0．05）且抑制INS-1细胞分泌胰岛素（t=9．592，P〈0．05）。结论c．pn感染对胰岛B细胞增殖和功能具有双向作用，可为2型糖尿病早期防治提供新思路。Objective To explore the effects of infection of Chlamydia pneumonia （C. pn） on the proliferation and function of pancreatic β cells. Methods C. pn had been cultured in the human laryngeal epidermoid carcinoma cell line（HEp-2 cells）. Then the pancreatic β cells （INS-1 cells） were infected by C. pn,the multiplicity of infection were 0.08 and 0.20, the time for intervention were 6 h and 24 h. The changes of cell morphology under the light microscopy and the immunofluoreseence monoclonal antibody stai- ning method were used to confirm the establishment of infection model. Under the infection of C. pn with dif- ferent nmhiplicity of infection （0,0.04,0.08,0.10,0.12,0.14,0.16,0.20） and different time （2,4,6,8, 16,24,36 h）, the methyl thiazolyl tetrazolium assay, the trypan blue and the flow cytomett7 were used to evaluate the effects of C. pn on the proliferation and function of INS-1 cells. Moreover, the low glucose （5.6 mmol/L） and high glucose （25 mmol/L）-stimulated insulin secretion of INS-1 cells,infected with C. pn, were measured through radioimmunoassay. Results Compared with normal control group, the cell density in- creased and the cell gap became more narrow in C. pn interventional group with a small dose （0.08） plus a short time （ 6 h ） . However , the cells became shrink and round , still agglomerated growing , in C. pn intervent -ional group with a large dose （0.20） plus a long time （24 h）. The immunofluorescent monoclonal antibody staining showed a specificity bright green fluorescent, confirming the C. pn inflectional cell model in vitro had been successfully established. Compared with normal control group, the infection of C. pn with a small dose （ 〈 0.20） plus a short time （ 〈 24 h） could induce the proliferation of INS-1 cells （ t = - 8. 907,P 〈 0.05 ） and increase the glucose-stimulated insulin secretion of INS-1 cells （ t = - 9. 186, P 〈 0.05 ） , but a large dose （〉0.20） plus a long time （ 〉24 h） could induce the apop

关 键 词：2型糖尿病 胰岛素抵抗 胰岛Β细胞 肺炎衣原体 

分 类 号：R5[医药卫生—内科学]