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分别于小鼠感染日本血吸虫后 0、 4、 6、 8和 12周 ,取脾淋巴细胞体外培养 ,进行细胞信号转导抑制试验 ,观察酪氨酸蛋白激酶 (TPK )、蛋白激酶C (PKC )和磷酯酰肌醇 3 激酶 (PI 3 K )特异性抑制剂 (Tyrphostin 2 5、D sphingosine和Wort mannin )分别特异性抑制和不同组合抑制TPK、PKC和PI 3K后 ,对小鼠脾淋巴细胞经虫卵可溶性抗原 (SEA )诱生IL 2、IFN γ和IL 4的表达水平及对Th1/Th2免疫偏移的影响。结果发现Tyrphostin 2 5对IL 2、IFN γ和IL 4水平的抑制作用均非常显著(P <0 0 1) ,D sphingosine主要影响IL 4的表达 (P <0 0 1) ,而Wortmannin则主要影响IFN γ的表达 (P <0 0 1) ,Tyrphostin 2 5和Wortmannin联合应用可完全阻断IL 2的表达及增强对IFN γ的抑制作用 ,Tyrphostin 2 5和D sphingosine联合应用可完全阻断IL 4的表达。对反映Th1/Th2免疫平衡的Th2分化指数分析表明 ,D sphingosine可使Th2免疫应答优势减弱 ,而Wort mannin则可使Th2免疫应答优势增强。研究结果表明 ,干预细胞信号转导可调节日本血吸虫特异性Th1/Th2细胞因子表达水平及Th1/Th2免疫偏移 ,为探索控制日本血吸虫卵肉芽肿病变的潜在新途径 ,提供了实验依据。
Splenic lymphocytes were cultured in vitro at 0, 4, 6, 8 and 12 weeks after infection with Schistosoma japonicum respectively. Signal transduction inhibition assay was performed to observe the effects of tyrosine kinase (PKK), protein kinase C (PKC) And PI3K specific inhibitors (Tyrphostin 2 5, Dsphingosine and Wort mannin), respectively, after specific inhibition of TPK, PKC and PI 3K inhibition of mouse splenic lymphocytes The expression levels of IL 2, IFNγ and IL 4 induced by soluble antigen of sera (SEA) and their effect on Th1 / Th2 immune offset. The results showed that the inhibitory effect of Tyrphostin 2 5 on IL 2, IFN γ and IL 4 levels was significant (P 0 01), D sphingosine mainly affected the expression of IL 4 (P 0 01), while Wortmannin The combination of Tyrphostin 25 and Wortmannin could completely block the expression of IL-2 and enhance the inhibition of IFN-γ, and the combination of Tyrphostin 25 and Dsphingosine could completely block the expression of IL-4 (P <0.01) expression. Analysis of the Th2 differentiation index reflecting the Th1 / Th2 immune balance showed that D sphingosine attenuated the predominance of the Th2 immune response, whereas Wort mannin enhanced the predominance of the Th2 immune response. The results show that intervention of cell signal transduction can regulate Schistosoma japonicum specific Th1 / Th2 cytokine expression levels and Th1 / Th2 immune bias, providing an experimental basis for exploring potential new ways to control Schistosoma japonicum egg granuloma lesions.