活动期强直性脊柱炎患者外周血单个核细胞CXCL16 mRNA和CXCL6 mRNA的表达及CXCL16对淋巴细胞增殖的影响

Expression of CXCL16 mRNA and CXCL6 mRNA in peripheral blood mononuclear cells of patients with active ankylosing spondylitis and its effect on lymphocyte proliferation

目的观察活动期强直性脊柱炎患者外周血单个核细胞CXC型趋化因子配体(CXC tchemokine ligand,CXCL)16和CXCL6的表达情况,探讨CXCL16对淋巴细胞增殖的影响。方法选择金华市中医医院2014年1月—2015年12月活动期强直性脊柱炎患者62例作为实验组,健康体检者62例作为对照组。测定血清CXCL16,外周血单个核细胞CXCL16 mRNA和CXCL6 mRNA水平、淋巴细胞增殖、培养上清液中RANKL水平。结果实验组患者血清CXCL16水平(2.47±0.97)ng/ml高于对照组(1.87±0.64)ng/ml(t=4.375,P=0.009),实验组CXCL16 mRNA和CXCL6 mRNA相对表达量(0.063±0.004、0.047±0.008)均高于对照组(0.034±0.003、0.020±0.005)(t=5.274、6.252,P=0.003、P<0.001)。在重组蛋白CXCL16作用下,实验组淋巴细胞增值率(169.23%±34.26%)和光密度值(0.038±0.002)均高于对照组(116.45%±23.54%、0.021±0.002)(t=6.847、6.035,均P<0.001),实验组淋巴细胞分泌RANKL水平(1.794±0.315)pmol/L高于对照组(0.957±0.264)pmol/L(t=5.264,P=0.002)。实验组中CXCL16组淋巴细胞p-ERK1/2、p-Raf-1蛋白表达量均高于其他3组(P=0.013、0.017、0.008;0.011、0.012、0.001)。实验组中CXCL16组S期和G_2/M期细胞比例高于其他3组(P=0.021、0.018、0.009;均P<0.001)。结论 CXCL16及其受体CXCL6可能参与强直性脊柱炎发病,其机制可能与CXCL16促进淋巴细胞增殖有关。

Objective To observe the expression of CXC tchemokine ligand （CXCL） 16 and CXCL6 in peripheral blood mononuclear cells of activities ankylosing spondylitis patient, to explore the impact of CXCL16 on lymphocyte proliferation. Methods Sixty-two cases of patients with activities ankylosing spondylitis were selected as the experimental group and 62 cases of healthy as control group in Jinhua City Chinese medicine hospital from January ,2014 to December,2015. The levels of serum CXCL16 ,the level of CXCL16 mRNA and CXCL6 mRNA in peripheral blood mononuclear cells ,lymphocyte proliferation, and the culture supernatant RANKL levels were detected. Results The serum CXCL16 level [ （2.47 ±0.97 ） ng/ml ] of experimental group was higher than that of control group [ （ 1.87 ±0.64 ） ng/ml ], t = 4. 375, P = 0. 009. The CXCL16 mRNA and CXCL6 mRNA relative expression （0. 063 ±0. 004,0. 047 ±0. 008 ） of experimental group were higher than that of control group （0.034 ±0.003,0. 020 ±0. 005 ） , t = 5. 274,6. 252, P = 0. 003, P 〈0. 001. In the role of recombinant CXCL16 protein,the lymphocyte proliferation rate[ （169.23 ±34.26）% ] and the optical density value （0.038 ±0. 002） of the experimental group were higher than that of control group [ （116.45 ±23.54）%, 0. 021 ±0. 002 ], t = 6. 847,6.035,P 〈 0.001. The levels of lymphatic cells secrete RANKL [ （ 1. 794 ± 0.315 ） pmol/L ] of experimental group was higher than that of control group （0. 957 ±0. 264 pmol/L） , t = 5. 264, P = 0. 002. The lymphocyte p-ERK1/2,p-Raf-1 protein expression of CXCL16 group were higher than other three groups （P = 0. 013,0. 017, 0. 008 ;0. 011,0. 012,0. 001 ）. The S phase and GJM phase cells ratio of CXCL16 group were higher than other three groups （ P = 0. 021,0.018,0. 009 ;P 〈 0. 001 ）. Conclusion CXCL16 and its receptor CXCL6 may involve the pathogenesis of ankylosing spondylitis ,its mechanism may be associated with CXCL16 promoting lymphocyte proliferation.