佐剂性关节炎大鼠细胞因子和淋巴细胞亚群变化与可铁宁的影响

Effects of cotinine on cytokines and leukomonocyte subgroup in rats with adjuvant-induced arthritis

目的:观察吸烟者体内尼古丁代替主要产物可铁宁在佐剂性关节炎动物模型抗炎和调节免疫方面的作用。方法:实验于2005-10/2006-09在南方医科大学南方医院检验科及实验动物中心完成。实验动物:SD大鼠50只。实验分组和给药:随机选取SD大鼠40只注射完全弗氏佐剂,致炎建立佐剂性关节炎模型,剩余的10只SD大鼠为正常组。将40只佐剂性关节炎大鼠随机分为炎症组10只,可铁宁低、中、高剂量组各10只,可铁宁各组于佐剂注射后1周开始灌胃给药,分别给0.5,1.5,4.5g/kg体质量剂量药物,连续7d。实验评估:致炎后28d开始进行运用ELISA及流式细胞术,检测用药后对白细胞介素1β、白细胞介素2,T细胞增殖等指标(CD4+,CD8+,CD4+/CD8+)的影响。结果:50只大鼠全部进入结果分析。①各组大鼠血清中白细胞介素1β、白细胞介素2含量:与炎症组相比,可铁宁高、中、低剂量各组白细胞介素1β含量明显下降[(101.4±11.24),(53.40±8.23),(60.34±5.97),(72.31±8.06)ng/L,P<0.05],白细胞介素2含量明显升高[(121.3±21.2),(195.9±51.6),(174.3±40.3),(169.7±53.2)ng/L,P<0.05],且作用有一定的剂量依赖性。②各组大鼠淋巴细胞亚群CD4+,CD8+,CD4+/CD8+变化:造模第28天,与正常组相比,炎症组的CD4+增加,CD8+明显降低(P<0.05),CD4+与CD8+的比值显著增加(P<0.05);与炎症组相比较,可铁宁高剂量组的CD4+显著降低(P<0.05);可铁宁各组的CD8+明显增加(P<0.05),CD4+与CD8+比值显著下降,呈剂量依赖性。结论:可铁宁可调节大鼠血清中异常的白细胞介素1及白细胞介素2含量,可铁宁在试验动物水平有抗炎和免疫调节的作用。

AIM： To observe the anti-inflammatory effects of cotinine, the main product of nicotine substitute, on adjuvant-induced arthritis in rats and its effect on regulating immunity. METHODS： The experiment was conducted in the Clinical Laboratory and Experimental Animal Center of Nanfang Hospital, Southern Medical University from October 2005 to September 2006. Forty SD rats of 50 were selected randomly and injected complete Freund＇s adjuvant to establish the models of adjuvant-induced arthritis; the left 10 rats were regarded as control group. The 40 rats with adjuvant arthritis were divided randomly into arthritis group, low, middle and high cotinine groups with 10 animals in each group. Three cotinine groups were injected 0.5, 1.5, 4.5 g/kg body mass cotinine one week after adjuvant injection for 7 days. On the 28^th day after modeling, the effects of cotinine on serum intedeukin-1β（IL-1β）, IL-2 and CD4^＋, CD8^＋, and CD4^＋/CD8^＋ leukomonocyte were measured with ELISA assays and flow cytometry method, respectively. RESULTS： Fifty rats entered the result analysis. （1）Compared with the arthritis group, the IL-1β levels of high, middle and low cotinine groups were significantly decreased [（101.4 ±11.24）, （53.40 ±8.23）, （60.34 ±5.97）, （72.31 ±8.06） ng/L, P 〈 0.05], and IL-2 levels were remarkably increased [（121.3±21.2）, （195.9±51.6）, （174.3±40.3）, （169.7±53.2） ng/L, P 〈 0.05]. The effect of cotinine was in a dose-dependent manner. （2）On day 28, CD4^＋ was increased, CD8^＋ decreased in t he arthritis group compared with the control group （P 〈 0.05）, and the ratio of CD4^＋/CD8^＋ was remarkably increased （P 〈 0.05）; compared with the arthritis group, CD4^＋ was increased, CD8^＋ decreased in three cotinine groups （P 〈 0.05）, and the ratio of CD4^＋/CD8^＋ was remarkably decreased, which was also in a dose-dependent manner. CONCLUSION： Cotinine could regulate the content of serum abnormal IL-1β and IL-2 in rats. It has immunosuppressive and anti-inflammatory effects at experimental animal level.