壮医针挑疗法对哮喘模型小鼠TSLP、OX40L及Th2优势分化应答的影响

Effects of needle-pricking therapy of Zhuang medicine on TSLP,OX40L and Th2-dominated differentiation immune response in mouse models of asthma

目的探讨壮医针挑疗法对哮喘模型小鼠胸腺基质淋巴细胞生成素(TSLP)、协同刺激分子配体(OX40L)及辅助性T细胞2(Th2)优势分化的影响和作用机制。方法将50只6~8周龄SPF级BALB/c小鼠分为对照组、模型组、针挑组、沙丁胺醇阳性对照组(阳性组)、假针挑组(假针组)5组(n=10)。应用卵清蛋白致敏及激发对模型组、针挑组、阳性组、假针组进行造模。针挑组在模型基础上应用壮医针挑疗法干预。干预1个周期后检测OX40LmRNA表达水平,血清IL-4、IL-5、IL-10、IL-13、IL-12水平以及TSLP表达。结果和对照组比较,模型组的TSLP蛋白、OX40LmRNA表达及IL-4、IL-5、IL-10、IL-13水平均升高而IL-12水平下降(P<0.05或P<0.01);和模型组比较,针挑组的TSLP蛋白、OX40LmRNA表达及IL-4、IL-5、IL-10、IL-13水平均降低而IL-12水平升高(P<0.05或P<0.01)。结论壮医针挑疗法能够降低TSLP蛋白和OX40LmRNA的过度表达、抑制Th2优势分化,减轻哮喘的炎症反应,推测这是壮医针挑疗法临床治疗哮喘的起效环节之一。

Objective To explore the changes of thymi c stromal lymphopoietin （TSLP）, costimulatory molecules ligand（OX40L） and T helper cell 2 （Th2）-dominated differentiation in mouse models of asthma after the treatment with needle-pricking of Zhuang medicine and to discuss its therapeutic mechanisms. Methods Fifty BALB/c mice aged from six to eight weeks old with SPF grade were randomly divided into five groups： a control group, a model group, and a needle-pricking group, salbutamol positive group and sham needle-prick- ing group with ten mice in each group. The mouse models of asthma of the model group, of the needle-pricking group, of the positive group and of the sham needle-pricking group were established by ovalbumin （OVA） sen- sitization and stimulating. The needle-pricking group was added with needle-pricking treatment. After a thera- peutic course was finished, the expression of OX40L mRNA was detected by RT-PCR, the levels of serum IL- 4, IL-5, IL-10, IL-13 and IL-12 were measured by ELISA, and the expression of TSLP protein was measured by Western blot. Results Compared with the control group, in the model group both the expressions of TSLP protein and OX40L mRNA and the levels of serum IL-4, IL-5, IL-10 and IL-13 increased significantly while the level of serum IL-12 reduced （ P〈0.05 or P〈0. 01）. The expressions of TSLP protein and OX40LmRNA and the levels of serum IL-4, IL-5, IL-10 and IL-13 reduced significantly while the level of ser- um IL-12 increased in the needle-pricking group, compared with model group, there were statistically significant differences （ P〈0.05 or P〈0.01）. Conclusion Needle-pricking therapy of Zhuang medicine can improve symptoms of asthma by reducing the overexpression of TSLP protein and OX40LmRNA and by inhibiting Th2-dominated differentiation, which was supposed to be one of the possible therapeutic mechanisms for asthma.