CAJ | 학술논문

目的观察特异性小干扰RNA (siRNA)沉默Cbl-b基因对于小鼠脾脏T淋巴细胞免疫活性影响.方法尼龙毛柱法分离T淋巴细胞,加入抗CD3抗体(1 mg/L)在体外模拟T淋巴细胞活化的第一信号,脂质体转染12、24、48 h后,流式细胞仪检测早、中、晚期活化标志分子CD69、CD25、CD71的表达变化；转染48 h后细胞计数试剂盒(CCK-8)试剂盒检测T淋巴细胞增殖程度.结果利用特异性Cbl-b siRNA能有效沉默T淋巴细胞Cbl-b基因,转染12 h后,转染组、对照组、空白组CD69分子表达阳性率分别为(46.78±4.89)％、(21.36±2.06)％、(25.09±2.27)％(P＜0.01)；转染24h后,转染组、对照组、空白组CD25分子表达阳性率分别为(19.27±1.84)％、(8.18±0.71)％、(9.25±0.65)％(P＜0.01)；转染48 h后,转染组、对照组、空白组CD71分子表达阳性率分别为(43.26 ±3.89)％、(23.99 ±1.97)％、(28.09 ±2.15)％ (P ＜0.01)；转染48 h后,CCK-8试剂盒检测转染组、对照组、空白组吸光度值分别为0.763±0.063、0.356±0.039、0.383±0.015(P＜0.01).结论利用特异性小干扰沉默Cbl-b基因能够增强小鼠脾脏T淋巴细胞免疫活性.
목적 관찰특이성소간우RNA (siRNA)침묵Cbl-b기인대우소서비장T림파세포면역활성영향.방법 니룡모주법분리T림파세포,가입항CD3항체(1 mg/L)재체외모의T림파세포활화적제일신호,지질체전염12、24、48 h후,류식세포의검측조、중、만기활화표지분자CD69、CD25、CD71적표체변화；전염48 h후세포계수시제합(CCK-8)시제합검측T림파세포증식정도.결과 이용특이성Cbl-b siRNA능유효침묵T림파세포Cbl-b기인,전염12 h후,전염조、대조조、공백조CD69분자표체양성솔분별위(46.78±4.89)％、(21.36±2.06)％、(25.09±2.27)％(P＜0.01)；전염24h후,전염조、대조조、공백조CD25분자표체양성솔분별위(19.27±1.84)％、(8.18±0.71)％、(9.25±0.65)％(P＜0.01)；전염48 h후,전염조、대조조、공백조CD71분자표체양성솔분별위(43.26 ±3.89)％、(23.99 ±1.97)％、(28.09 ±2.15)％ (P ＜0.01)；전염48 h후,CCK-8시제합검측전염조、대조조、공백조흡광도치분별위0.763±0.063、0.356±0.039、0.383±0.015(P＜0.01).결론 이용특이성소간우침묵Cbl-b기인능구증강소서비장T림파세포면역활성.
Objective To investigate the effect of specific small interfering RNA (siRNA) silencing the expression of casitas B cell lymphoma-b (Cbl-b) gene on the immune activity of the mouse spleen T lymphocytes.Methods T lymphocytes were isolated by Nylon Wool Fiber Columns.By using anti-CD3 antibodies (1 mg/L) as the first signal of T lymphocyte activation in vitro,and at 12,24 and 48 h after transfecting by liposome,the expression of CD69,CD25 and CD71 was detected and analyzed by using fluorescence conjugated monoclonal antibodies and flowcytometry.At 48 h after transfection,the proliferation of the T lymphocyte was measured with CCK-8 kit.Results Specific Cbl-b siRNA could effectively silence the expression of Cbl-b gene.At 12 h after transfection,CD69 molecular expression levels in transfection group,negative control group and blank group were (46.78 ±4.89)％,(21.36 ±2.06)％,and (25.09 ±2.27)％ respectively (P ＜ 0.01).At 24 h after transfection,CD25 molecular expression levels in transfection group,negative control group and blank group were (19.27 ± 1.84)％,(8.18 ±0.71)％,and (9.25 ± 0.65) ％ respectively (P ＜ 0.01).At 48 h after transfection,CD71 molecular expression levels in transfection group,negative control group and blank group were (43.26 ± 3.89)％,(23.99 ±1.97) ％,and (28.09 ± 2.15)％ respectively (P ＜ 0.01).At 48 h after transfection,A values detected by CCK-8 kit in transfection group,control group,and blank group were 0.763 ± 0.063,0.356 ± 0.039,and 0.383 ± 0.015 respectively (P ＜ 0.01).Conclusion Specific siRNA silencing Cbl-b gene can markedly promote the immune activity of mouse T lymphocytes.