CXCL16在胶原诱导性关节炎小鼠中的表达及对淋巴细胞增殖活化的影响

Expression of chemokine CXCL16 in murine collagen-induced arthritis and the effects on the proliferation of lymphocytes

目的了解CXCLl6在胶原诱导性关节炎（CIA）小鼠发病中的作用。方法以8只正常小鼠为对照，采用酶联免疫吸附试验（ELISA）和反转录一聚合酶链反应（RT—PCR）法，分别检测CXCLl6在12只CIA小鼠外周血中的蛋白水平及病变滑膜组织中的mRNA水平，再采用CCK-8法、RT-PCR和ELISA，分别检测不同浓度的CXCLl6重组蛋白（0，100，200，400，800ng／m1）刺激鼠脾淋巴细胞的增殖情况、上清中自细胞介素（IL）-2和干扰素一吖水平及RANKLmRNA表达水平。采用t检验和单因素方差分析进行统计分析。结果①与对照组相比，CIA小鼠外周血CXCL16[分别为（72±8）和（127±10）pg／m1，P〈0．05]和滑膜CXCLl6mRNA水平（分别为0．214±0．007和0．375±0．009，P〈0．01）均显著增高。②CXCLl6（200、400、800ng／m1）作用于CIA小鼠淋巴细胞，其增殖能力明显高于CXCLl6（0n/ml），吸光度（A）值分别为0．51±0．06、0．56±0．05、O．55±0．04、0．41±0．04（P〈O．05）。并且CXCLl6对CIA组小鼠淋巴细胞的增殖刺激能力明显高于其对健康对照组小鼠淋巴细胞（P均〈0．05）。③核因子KB受体活化因子配体（RANKL）mRNA、IL-2和干扰素-1水平在CIACXCLl6（400、800ng/ml）组均明显高于CXCLl6（0ng／m1）组（P〈0．01）。结论CXCLl6可能通过活化淋巴细胞在CIA发病中发挥重要作用。

Objective To investigate the effect of CXCL16 on the development of murine collagen- induced arthritis （CIA）. Methods CXCLI6 mRNA of the involved synovium and serum CXCL16 protein were determined respectively by reverse transcription-polymerase chain reaction （RT-PCR） and enzyme linked immunosorbent assay （ELISA） in murine collagen-induced arthritis. The proliferation of lymphocytes from murine spleen and the level of RANKL mRNA, stimulated by CXCL16 at different concentrations （0, 100, 200, 400, 800 ng/ml）, was detected respectively by CCK-8 and RT-PCR, then the level of IL-2 and IFN-~/ in culture supernatant was detected by ELISA. Comparisons between groups were tested by t test and one-way ANOVA analysis. Results The serum CXCLI6 [ （127±10） vs （72±8） pg/ml, P〈0.05 ] and synovial CXCLI6 mRNA （0.214±0.007 vs 0.375＋0.009, P〈0.01） in CIA were all significantly higher than those in normal controls. The proliferation of CXCL16 （200, 400, 800 ng/ml） in CIA mouse lymphocytes, was significantly higher than that of CXCL16 （0 ng/ml） （0.51±0,06, 0.56±0.05, 0.55±0.04, （0.41±0.04, P〈 0.05）. And CXCL16 on the CIA stimulated lymphocyte proliferation was significantly higher than controls on normal lymphocytes （P〈0.05）. Compared with blank control group, the expression of IL-2, IFN-γ and RANKL mRNA of CIA CXCL16 （400, 800 ng/ml） groups was higher significantly （P〈0.01）. Conclusion CXCL16 plays an important role in the development of routine CIA by activating lymphocytes.