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目的通过研究活化T细胞核因子(NFAT)在邻苯二甲酸二(2-乙基已基)酯(DEHP)影响白细胞介素-4(IL-4)表达中的作用,探讨DEHP的免疫毒性机制。方法以佛波酯(PMA)和离子霉素(Ion)为激活剂,用10、50μmol/L DEHP染毒激活的脾淋巴细胞4 h,同时用0.5μmol/L钙调神经磷酸酶抑制剂他克莫司(FK506)进行干预,最后分别检测IL-4、NFATp、NFATc mRNA表达与NFATp和NFATc蛋白表达。结果以10 ng/mL PMA+0.5mg/mL Ion为激活剂时,激活组的IL-4 mRNA表达相对值为(2.53±0.42),50μmol/L DEHP染毒组的相对值为(18.21±1.67),差异有统计学意义(P<0.01);而FK506干预组的IL-4 mRNA表达相对值为(9.85±1.24),与50μmol/L DEHP染毒组比较,差异有统计学意义(P<0.01);50μmol/L DEHP染毒组的NFATc mRNA表达相对值为(2.00±0.32),激活组的相对值为(1.37±0.10),差异有统计学意义(P<0.05);同时DEHP促进NFATc蛋白在细胞质和细胞核里表达;染毒4 h后,与激活组NFATp mRNA表达相对值(2.51±0.45)比较,10、50μmol/L DEHP染毒组的相对值分别为(5.14±0.28)和(5.38±0.60),差异均有统计学意义(P<0.01);但DEHP降低了细胞质和细胞核里NFATp蛋白表达。结论 NFAT在DEHP调控IL-4表达中具有重要作用。
OBJECTIVE: To investigate the effect of DEAT on the expression of interleukin-4 (IL-4) by studying the effect of NFAT on the immunotoxicity of DEHP . Methods Activation of splenic lymphocytes by phorbol ester (PMA) and ionomycin (Ion) for 4 h with 10, 50 μmol / L DEHP was performed with 0.5 μmol / L calcineurin inhibitor (FK506) for intervention. Finally, the expression of IL-4, NFATp, NFATc mRNA and the expression of NFATp and NFATc protein were detected respectively. Results The relative expression of IL-4 mRNA in the activated group was (2.53 ± 0.42) and that in the 50 μmol / L DEHP-treated group was (18.21 ± 1.67) with 10 ng / mL PMA + 0.5 mg / ), The difference was statistically significant (P <0.01), while the relative value of IL-4 mRNA expression in FK506 intervention group was (9.85 ± 1.24), which was significantly different from that in 50μmol / L DEHP exposure group (P < 0.01). The relative expression of NFATc mRNA in DEHP-treated group was (2.00 ± 0.32) and that in activated group was (1.37 ± 0.10), the difference was statistically significant (P <0.05) The protein expression in cytoplasm and nucleus was significantly higher than that in 10 and 50μmol / L DEHP groups (P <0.05). The relative values of NFATp mRNA expression (2.51 ± 0.45) 5.38 ± 0.60), the differences were statistically significant (P <0.01); however, DEHP reduced NFATp protein expression in cytoplasm and nucleus. Conclusion NFAT plays an important role in the regulation of IL-4 expression by DEHP.