CAJ | 학술논문

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(16.1586 pg/mL),随后逐渐降低但仍高于正常水平(3.4146 pg/mL P =0.017);第10w sIL-13Rα2的分泌达到高峰(4827.426 pg/mL),以后逐渐减低,但仍高于正常水平(4057.112 pg/mL P =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.
목적 연구재성섬유세포NIH-3T3중IL-13가용성수체sIL-13Rα2대IL-13적억제작용,위진일보연구sIL-13Rα2대일본혈흡충감염소서체내간섬유화적치료작용전정기출.방법 용ELISA화RT-PCR검측감염일본혈흡충적BALB/c소서0、6、8、10화12w불동감염시기간장조직IL-13화sIL-13Rα2표체화전록수평.구건sIL-13Rα2표체질립전염성섬유세포NIH-3T3,용IL-13(50ng/mL)자격전염후성섬유세포NIH-3T3,용RT-PCR화Western blotting분별검측해세포분비적Ⅰ형효원.결과 감염후소서간장육아종조직중IL-13화sIL-13Rα2 적단백표체수평수감염시간적연장이축점증고,제8wIL-13수평체도고봉(16.1586 pg/mL),수후축점강저단잉고우정상수평(3.4146 pg/mL P =0.017);제10w sIL-13Rα2적분비체도고봉(4827.426 pg/mL),이후축점감저,단잉고우정상수평(4057.112 pg/mL P =0.021).IL-13 화sIL-13Rα2적mRNA전록추세화ELISA 검측결과 상부합,균수감염시간적연장이증고,분별재제8w화제10w체도최고봉,수후축점강저단잉고우정상조소서(P =0.033; P =0.025).실험조(sIL-13Rα2=2mg/mL)Ⅰ형효원mRNA전록수평교정상대조조감저8.83%(P =0.012);단백수평교대조조감저7.41%(P =0.031).결론 sIL-13Rα2재NIH-3T3세포중대IL-13유억제작용,제시sIL-13Rα2재치료혈흡충병간섬유화중구유잠재개치.
To determine the inhibition of IL-13 by recombinant sIL-13Rα2 in NIH-3T3 fibroblast cells for its potential therapeutic value in hepatic fibrosis caused by Schistosoma japanicum in mice . IL-13 and sIL-13Rα2 from liver of BALB/c mice infected with S.japonicum at different infection time (weeks 0,6,8,10 and 12) were analyzed by ELISA and RT-PCR. The recombinant sIL-13Rα2 expression plasmidwas constructed, followed by transfection into NIH-3T3 fibroblast cells. TypeⅠcollagen produced by NIH-3T3 cells were examined by RT-PCR and Western blotting. It was demonstrated that the expression of IL-13 increased gradually after infection, reached peak density (16.1586 pg/mL)at week 8 and then reduced but was still higher than the level of control mice(3.4146 pg/mL;P =0.017 ). The secretion of sIL-13R α2 reached to its peak 10 weeks after infection(4827.426 pg/mL)and then reduced slowly but still higher than normal(4057.112 pg/mL; P=0.021). Meanwhile, the changes in mRNA level of IL-13 and sIL-13R α2 were coincided with that examined by ELISA. Both IL-13 and sIL-13Rα2 reached their peak density (P=0.033) at week 8 and 10 (P=0.025) respectively, and they were followed by a slower degree of decrease. The sIL-13Rα2 could significantly inhibit the effect of IL-13 on NIH-3T3 fibroblast cells, showing decreased mRNA level(P =0.012)and protein level of typeⅠcollagen compared with normal groups(P =0.031). It is concluded that the sIL-13Rα2 can inhibit the effect of IL-13 on NIH-3T3 fibroblast cells which leads to a reduced production of typeⅠcollagen, demonstrating its potential therapeutic value in hepatic fibrosis of schistosomiasis.