独一味胶囊对复发性口腔溃疡大鼠血清TNF-α、IL-2和IL-6水平的影响

Effect of the Duyiwei capsules on serum TNF-α,IL-2 and IL-6 levels in rats with recurrent oral ulcer

目的探讨独一味胶囊对复发性口腔溃疡(ROU)大鼠血清肿瘤坏死因子(TNF-α)、白介素2(IL-2)及白介素6(IL-6)水平的影响。方法 SPF级雄性SD大鼠40只,采用注射完全弗氏佐剂法建立ROU大鼠模型,随机分为4组(低、中、高剂量组及对照组),每组10只。低、中、高剂量组大鼠以1mL/100g灌服低、中、高剂量独一味胶囊溶液(独一味胶囊内容物以无菌生理盐水配置浓度为5%、10%、15%的溶液),对照组灌服1mL/100g无菌生理盐水。各组分别在给药1、3、7d后,对比各组大鼠口腔溃疡数目、溃疡面积,然后尾静脉采血,采用酶联免疫吸附实验(ELISA)检测各组血清TNF-α、IL-2及IL-6水平,并对比各组数据;给药7d后处死大鼠取溃疡组织,采用免疫印迹试验检测各组口腔溃疡组织中TNF-α、IL-2及IL-6蛋白相对表达量,并对比各组数据。结果低、中、高剂量组大鼠给药后不同时刻溃疡数目、溃疡面积、血清TNF-α、IL-2及IL-6水平组间、时间及交互比较,低、中、高剂量组大鼠给药7d后溃疡黏膜组织TNF-α、IL-2及IL-6蛋白相对表达量比较,差异有统计学意义(P<0.05)。低、中、高剂量组给药后不同时刻溃疡数目、溃疡面积、血清TNF-α、IL-6水平及给药7d后溃疡黏膜组织中TNF-α、IL-6蛋白相对表达量均显著低于对照组(P<0.05),且低、中、高剂量组均随时间及剂量依赖性降低(P<0.05);低、中、高剂量组给药后不同时刻血清IL-2水平及给药7d后溃疡黏膜组织中IL-2蛋白相对表达量均显著高于对照组(P<0.05),且3实验组均随时间及剂量依赖性升高(P<0.05);对照组大鼠给药后不同时刻溃疡数目、溃疡面积、TNF-α、IL-2、IL-6血清水平及给药7d后口腔溃疡黏膜组织中TNF-α、IL-2、IL-6蛋白相对表达量差异无统计学意义(P>0.05)。结论独一味胶囊可显著减少大鼠口腔溃疡数目及溃疡面积,加速大鼠口腔溃疡愈合,推测与独一味胶囊显著下调TNF-α、IL-6在血清及口腔溃疡黏膜组织中的表达及上调IL-2的表达有关。

Objective To explore the effects of the Duyiwei capsules on tumor necrosis factor-α（TNF-α）, interleukin-2 （IL-2） and interleukin-6 （IL-6） levels of serum in rats with recurrent oral ulcer （ROU）. Methods There were 40 males SD rats with SPF, and the rats model of ROU were established by injection of complete Freund＇s adjuvant method, which were randomly divided into 4 groups, 10 in each group. The rats of the low, medium and high dose group were given 1 mL/100 g of different concentrations of the Duyiwei capsules solution, which were prepared by using sterile saline solution with concentration of 5%, 10% and 15% of the Duyiwei capsules contents. The numbers and areas of oral ulcer of rats were observed and compared after 1, 3 and 7 days. The TNF-α, IL-2 and IL-6 levels of serum at 1, 3 and 7 days were detected by enzyme-linked immunosorbent assay （ELISA）. The rats were executed after 7 days and the ulcer tissues were taken. The relative expression of TNF-α, IL-2 and IL-6 proteins of the tissues were detected by Western Blotting. Results There were significant differences in the numbers and areas of oral ulcer, the serum levels of TNF-α, IL-2 and IL-6 among the 3 doses groups, moments and interactions （P 〈0.05）. The protein levels of TNF-α, IL-2 and IL-6 in the mucosal tissues after 7 days of the 3 doses groups were significantly different （P 〈0.05）. The numbers and areas of oral ulcer, the serum levels of TNF-α and IL-6 at 1, 3 and 7 days and the protein levels of TNF-α and IL-6 in the mucosal tissues after 7 days of the 3 doses groups were significantly lower than those of the control group （P 〈0.05）, every dose group showed a significant decrease with time and dose-dependence （P 〈0.05）. The serum levels of IL-2 at 1, 3 and 7 days and the protein levels of IL-2 in the mucosal tissues after 7 days of the 3 doses groups were significantly higher than those of the control group （P 〈0.05）, every dose group showed a significant increase with time and dose-dependence （P 〈0.05）. There were no significant differences between the numbers and areas of oral ulcer, the serum levels of TNF-α, IL-2 and IL-6 after drug delivery of 1, 3 and 7 days and the protein levels of TNF-α, IL-2 and IL-6 in the mucosal tissues after 7 days of the 3 dose groups （P 〉0.05）. Conclusion The Duyiwei capsules can significantly reduce the numbers and areas of oral ulcer of rats, also accelerate the healing of oral ulcer in rats, and it may be related to the levels of the serum and protein of TNF-α and IL-6 down-regulation and the serum and protein of IL-2 up-regulation by the Duyiwei capsules.