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目的研究在成纤维细胞NIH-3T3中IL-13可溶性受体sIL-13Rα2对IL-13的抑制作用,为进一步研究sIL-13Rα2对日本血吸虫感染小鼠体内肝纤维化的治疗作用奠定基础。方法用ELISA和RT-PCR检测感染日本血吸虫的BALB/c小鼠0、6、8、10和12w不同感染时期肝脏组织IL-13和sIL-13Rα2表达和转录水平。构建sIL-13Rα2表达质粒转染成纤维细胞NIH-3T3,用IL-13(50ng/mL)刺激转染后成纤维细胞NIH-3T3,用RT-PCR和Western blotting分别检测该细胞分泌的Ⅰ型胶原。结果感染后小鼠肝脏肉芽肿组织中IL-13和sIL-13Rα2的蛋白表达水平随感染时间的延长而逐渐增高,第8wIL-13水平达到高峰(16.1586pg/mL),随后逐渐降低但仍高于正常水平(3.4146pg/mLP=0.017);第10wsIL-13Rα2的分泌达到高峰(4827.426pg/mL),以后逐渐减低,但仍高于正常水平(4057.112pg/mLP=0.021)。IL-13和sIL-13Rα2的mRNA转录趋势和ELISA检测结果相符合,均随感染时间的延长而增高,分别在第8w和第10w达到最高峰,随后逐渐降低但仍高于正常组小鼠(P=0.033;P=0.025)。实验组(sIL-13Rα2=2mg/mL)Ⅰ型胶原mRNA转录水平较正常对照组减低8.83%(P=0.012);蛋白水平较对照组减低7.41%(P=0.031)。结论sIL-13Rα2在NIH-3T3细胞中对IL-13有抑制作用,提示sIL-13Rα2在治疗血吸虫病肝纤维化中具有潜在价值。
Objective To study the inhibitory effect of IL-13 soluble receptor sIL-13Rα2 on IL-13 in fibroblasts NIH-3T3, and lay a foundation for the further study on the therapeutic effect of sIL-13Rα2 on hepatic fibrosis induced by Schistosoma japonicum in mice. Methods The expression and transcript levels of IL-13 and sIL-13Rα2 in liver tissues of BALB / c mice infected with Schistosoma japonicum at 0, 6, 8, 10 and 12w infection were detected by ELISA and RT-PCR. The NIH-3T3 fibroblasts were transfected with sIL-13Rα2 expression plasmid and NIH-3T3 fibroblasts were stimulated with IL-13 (50ng / mL). The expression of type Ⅰ was detected by RT-PCR and Western blotting respectively. collagen. Results The protein expression levels of IL-13 and sIL-13Rα2 in the hepatic granuloma tissue of infected mice increased gradually with the prolongation of infection time, reached the peak at 16th day (16.1586pg / mL), and then decreased gradually but still high At the normal level (3.4146 pg / mL P = 0.017), the secretion of 10wsIL-13Rα2 peaked (4827.426 pg / mL) and then decreased gradually but remained above the normal level (4057.112 pg / mL P = 0.021). The mRNA transcription trends of IL-13 and sIL-13Rα2 were in accordance with those of ELISA, which were both increased with the prolongation of infection time and reached the peak at 8th and 10th weeks respectively, then decreased gradually but still higher than that of normal mice P = 0.033; P = 0.025). The transcription level of type Ⅰ collagen in experimental group (sIL-13Rα2 = 2mg / mL) was reduced by 8.83% (P = 0.012) compared with that in normal control group, and decreased by 7.41% (P = 0.031). Conclusion sIL-13Rα2 inhibits IL-13 in NIH-3T3 cells, suggesting that sIL-13Rα2 has potential value in the treatment of liver fibrosis induced by schistosomiasis.