论文部分内容阅读
目的:探讨CCL20与急性病毒性心肌炎(AVMC)患者Th17细胞趋化性的关系。方法:选择11例AVMC患者为研究对象,同时选取10例健康人作为对照。利用siRNA技术干预AVMC患者外周血巨噬细胞,将其分为A(不接受siRNA转染)、B(接受CCL20 siRNA转染)、C(阴性对照,接受scramble siRNA转染)亚组。应用ELISA法检测AVMC组患者血清CCL20及其巨噬细胞体外分泌CCL20的水平,趋化实验和流式细胞学结合观察体外不同水平CCL20对Th17细胞的趋化能力及表达CCL20受体CCR6的Th17细胞比例的影响,实时定量PCR法检测趋化板下室细胞表面CCR6mRNA的表达。结果:与健康对照组[(2.78±0.64)μg/L]相比,AVMC组患者体内血清CCL20水平[(11.36±2.45)μg/L]明显升高,P<0.05。体外实验中,B亚组巨噬细胞培养上清CCL20含量、趋化实验募集的Th17细胞比例、表达CCR6的Th17细胞比例和CCR6mRNA相对表达水平[(0.94±0.08)ng/L、(6.86±0.59)%、(2.56±0.78)%、(0.36±0.08)]均低于A亚组[(2.57±0.25)ng/L、(9.74±0.77)%、(4.55±0.84)%、(1.00±0.15)]和C亚组[(2.08±0.17)ng/L、(9.16±0.82)%、(4.12±0.43)%、(0.88±0.17)],均P<0.05,而A、C亚组间各参数差异均无统计学意义。结论:AVMC患者中Th17细胞趋化性与巨噬细胞分泌的CCL20有关。
Objective: To investigate the relationship between CCL20 and Th17 cell chemotaxis in patients with acute viral myocarditis (AVMC). Methods: Eleven patients with AVMC were selected as study subjects, and 10 healthy individuals were selected as control. Peripheral blood macrophages were treated with siRNA technique and divided into A (not transfected with siRNA), B (transfected with CCL20 siRNA) and C (negative control, transfected with scramble siRNA) subgroups. The levels of CCL20 and CCL20 secreted by macrophages in vitro in patients with AVMC were detected by ELISA. Chemotaxis assay and flow cytometry were used to observe the chemotaxis ability of CCL20 to Th17 cells in vitro and Th17 cells expressing CCL20 receptor CCR6 Ratio of real-time quantitative PCR method to detect chemotactic platelet cell surface CCR6mRNA expression. Results: Compared with healthy controls [(2.78 ± 0.64) μg / L], serum CCL20 levels in patients with AVMC [(11.36 ± 2.45) μg / L] were significantly increased, P <0.05. In vitro experiments, the content of CCL20 in supernatant of macrophage culture in group B, the proportion of Th17 cells recruited in chemotaxis experiment, the proportion of Th17 cells expressing CCR6 and the relative expression level of CCR6 mRNA [(0.94 ± 0.08) ng / L, (6.86 ± 0.59 ), (2.56 ± 0.78)% and (0.36 ± 0.08), respectively, were lower than those in A subgroup (2.57 ± 0.25 ng / L, 9.74 ± 0.77%, 4.55 ± 0.84% ) And C subgroups [(2.08 ± 0.17) ng / L, (9.16 ± 0.82)%, (4.12 ± 0.43)%, (0.88 ± 0.17) No significant differences in parameters were found. Conclusion: The chemotaxis of Th17 cells in AVMC patients is related to CCL20 secreted by macrophages.