Objective:To explore the effects of down-regulated tryptase expression in mast cells on the synthesis and release of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) of vascular endothelial cells. Methods...Objective:To explore the effects of down-regulated tryptase expression in mast cells on the synthesis and release of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) of vascular endothelial cells. Methods: Tryptase-siRNA (small-interfering RNA) vector was constructed to inhibit tryptase expression in P815 cells. The medium of P815 cells treated by the tryptase-siRNA (RNAi-P815 group) or pure vector (P815 group) was collected and used to culture bEnd.3 cells. The mes-senger RNAs (mRNAs) of IL-6 and TNF-α in bEnd.3 cells and their protein levels in the medium were measured by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Results: IL-6 and TNF-α mRNAs in bEnd.3 cells cultured in RNAi-P815-conditioned medium decreased significantly compared to those in P815-conditioned medium. Consistently, IL-6 and TNF-α protein levels in the medium of bEnd.3 of RNAi-P815 group were lower than those of P815 group. Conclusion: Reduced tryptase expression significantly inhibited the synthesis and release of IL-6 and TNF-α in vascular endothelial cells. RNA interference targeting tryptase expression may be a new anti-inflammatory strategy for vascular diseases.展开更多
目的探讨γ干扰素(IFN-γ)对P815小鼠肥大细胞以及小鼠髓源性肥大细胞(BMMC)表达组织蛋白酶S(CTSS)的影响。方法用(10、25、50)ng/m L IFN-γ分别刺激P815细胞及原代培养的小鼠BMMC,利用实时定量PCR、ELISA分别检测P815细胞及BMMC CTSS...目的探讨γ干扰素(IFN-γ)对P815小鼠肥大细胞以及小鼠髓源性肥大细胞(BMMC)表达组织蛋白酶S(CTSS)的影响。方法用(10、25、50)ng/m L IFN-γ分别刺激P815细胞及原代培养的小鼠BMMC,利用实时定量PCR、ELISA分别检测P815细胞及BMMC CTSS的mRNA和蛋白水平。用25 ng/m L IFN-γ处理P815细胞及小鼠BMMC,P815细胞分别处理6、12、18、24、48、72 h,BMMC分别处理12、24、48 h,重复上述检测步骤。结果 IFN-γ可增加P815细胞及BMMC CTSS的mRNA和蛋白水平,并具有一定的时间、剂量依赖关系。结论 IFN-γ可促进小鼠肥大细胞表达CTSS。展开更多
基金Project (No. 30470689) supported by the National Natural ScienceFoundation of China
文摘Objective:To explore the effects of down-regulated tryptase expression in mast cells on the synthesis and release of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) of vascular endothelial cells. Methods: Tryptase-siRNA (small-interfering RNA) vector was constructed to inhibit tryptase expression in P815 cells. The medium of P815 cells treated by the tryptase-siRNA (RNAi-P815 group) or pure vector (P815 group) was collected and used to culture bEnd.3 cells. The mes-senger RNAs (mRNAs) of IL-6 and TNF-α in bEnd.3 cells and their protein levels in the medium were measured by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Results: IL-6 and TNF-α mRNAs in bEnd.3 cells cultured in RNAi-P815-conditioned medium decreased significantly compared to those in P815-conditioned medium. Consistently, IL-6 and TNF-α protein levels in the medium of bEnd.3 of RNAi-P815 group were lower than those of P815 group. Conclusion: Reduced tryptase expression significantly inhibited the synthesis and release of IL-6 and TNF-α in vascular endothelial cells. RNA interference targeting tryptase expression may be a new anti-inflammatory strategy for vascular diseases.
