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目的研究THP-1细胞受肺炎链球菌(SP)刺激后细胞间黏附分子1(ICAM-1)分泌水平的变化,并探讨不同来源SP对此影响的差异。方法从温州医学院附属第二医院住院患者分离、搜集23株SP,和ATCC49619一起进行培养,并调浓度至1.0 McFarland。收集经传代、培养的THP-1细胞,调浓度至4.0×108/L,取1.0 mL加入24孔细胞培养板中,再向其中加入100μL浓度为1.0 McFarland的SP,置37℃5%CO2环境分别孵育4和8 h,吸出细胞悬液,离心取上清液。以ELISA法检测上清液中ICAM-1的浓度。以SPSS 17.0软件对检测结果进行统计分析。结果 SP刺激THP-1细胞4、8 h后ICAM-1的浓度均高于相应的空白对照。血源性SP刺激THP-1细胞4、8 h后ICAM-1的浓度与相应的痰源性SP间差异均无统计学意义。血源性/痰源性SP刺激THP-1细胞8h后ICAM-1的浓度均高于其刺激4 h后的浓度。血源性/痰源性SP刺激THP-1细胞4 h后ICAM-1的浓度均高于ATCC49619菌株刺激4 h后相应的浓度,而在刺激8 h后二者间差异无统计学意义。结论 SP可使THP-1细胞分泌ICAM-1增加,但其浓度变化与刺激的时间有关。血源性和痰源性SP刺激THP-1细胞分泌ICAM-1的变化比较差异无统计学意义,而二者和ATCC49619菌株间差异有统计学意义。
Objective To investigate the changes of ICAM-1 secretion in THP-1 cells stimulated by Streptococcus pneumoniae (SP), and to explore the differences of the effects of different sources of SP on THP-1 cells. Methods Twenty-three SP strains were collected from inpatients in the Second Affiliated Hospital of Wenzhou Medical College and cultured with ATCC49619 and adjusted to 1.0 McFarland. The passaged and cultured THP-1 cells were collected and adjusted to a concentration of 4.0 × 10 8 / L. 1.0 mL of the THP-1 cells were added to a 24-well cell culture plate, and 100 μL of SP at 1.0 McFarland After 4 and 8 h incubation, the cell suspension was aspirated and the supernatant was centrifuged. The concentration of ICAM-1 in supernatant was detected by ELISA. SPSS 17.0 software for statistical analysis of test results. Results The concentration of ICAM-1 in THP-1 cells stimulated by SP for 4 and 8 h were all higher than that of corresponding blank control. No significant difference was found between the concentration of ICAM-1 and the corresponding sputum-derived SP at 4 and 8 h after the blood-derived SP stimulated THP-1 cells. The concentration of ICAM-1 in blood-stimulated / sputum-derived SP stimulated THP-1 cells for 8 h was higher than that of 4 h after stimulation. The concentration of ICAM-1 in THP-1 cells stimulated by blood-derived / phlegm-derived SP for 4 h was higher than that of ATCC49619 for 4 h. However, there was no significant difference between the two groups after stimulation for 8 h. Conclusion SP can increase the secretion of ICAM-1 in THP-1 cells, but its concentration changes with the time of stimulation. There was no significant difference in the changes of ICAM-1 secreted by THP-1 cells between blood-derived and sputum-derived SPs, but there was a significant difference between them and ATCC49619.