论文部分内容阅读
AIM To determine the functional role of mi R-490-5p in mast cell proliferation and apoptosis,and in the mast cell tryptase/PAR-2 signal pathway.METHODS The 3rd generation of lentivirus vector systems containing enhanced green fluorescent protein(EGFP)(Ruisai Inc.,Shanghai,China),which acts as a reporter gene was used to construct the mmu-mi R-490-5p lentivirus expression vector p EGFP-antagomi R-490-5p,and the lentivirus vector p EGFP-negative was used as a negative control.The stably transfected mast cell line p815 was then constructed.GFP positive cells were successfully transfected cells.We determined the expression of mi R-490-5p in p815 mast cells before and after transfection using quantitative real-time PCR(q RT-PCR).In addition,after transduction with the lentivirus vectors,the role of mi R-490-5p in mast cell proliferation and apoptosis was investigated using the CCK-8 assay and flow cytometry,respectively.The m RNA levels of tryptase and PAR-2 were detected by q RT-PCR and the protein levels were detected by Western blot.RESULTS The inhibition of mi R-490-5p expression promoted apoptosis and inhibited proliferation of p815 mast cells.The m RNA levels of tryptase and PAR-2 were significantly increased after transfection comparedwith the control group,tryptase(P=0.721,normal vs null;P=0.001,si RNA vs normal;P=0.002,si RNA vs null)and PAR-2(P=0.027,si RNA vs null;P=0.353,normal vs null;P=0.105,si RNA vs normal).The protein levels of tryptase and PAR2 were slightly higher in the si RNA group than those in the control group,but the difference was not statistically significant(P>0.05).CONCLUSION mi R-490-5p plays a vital role in the pathogenesis of irritable bowel syndrome by affecting mast cell proliferation and apoptosis;with down-regulation of mi R-490-5p,the m RNA level of mast cell tryptase and PAR-2 increased,and the protein level increased,but the difference was not statistically significant.
AIM To determine the functional role of mi R-490-5p in mast cell proliferation and apoptosis, and in the mast cell tryptase / PAR-2 signaling pathway. METHODS The 3rd generation of lentivirus vector systems containing enhanced green fluorescent protein (EGFP) ( Ruisai Inc., Shanghai, China), which acts as a reporter gene was used to construct the mmu-mi R-490-5p lentivirus expression vector p EGFP-antagomi R-490-5p, and the lentivirus vector p EGFP-negative was used as a negative control. stp transfected mast cell line p815 was then constructed. GFP positive cells were successfully transfected cells. We determined the expression of mi R-490-5p in p815 mast cells before and after transfection using quantitative real-time PCR (q RT-PCR). In addition, after transduction with the lentivirus vectors, the role of mi R-490-5p in mast cell proliferation and apoptosis was investigated using the CCK-8 assay and flow cytometry, respectively. m RNA levels of tryptase and PAR-2 were detected by q RT-PCR and th e protein levels were detected by Western blot .RESULTS The inhibition of mi R-490-5p expression promoted apoptosis and inhibited proliferation of p815 mast cells. The m RNA levels of tryptase and PAR-2 were significantly increased after transfection comparedwith the control group, P <0.001, si RNA vs normal; P = 0.002, si RNA vs null) and PAR-2 (P = P = 0.105, si RNA vs. normal). The protein levels of tryptase and PAR2 were slightly higher in the si RNA group than those in the control group, but the difference was not included significant (P> 0.05) .CONCLUSION mi R-490 -5p plays a vital role in the pathogenesis of irritable bowel syndrome by affecting mast cell proliferation and apoptosis; with down-regulation of mi R-490-5p, the m RNA level of mast cell tryptase and PAR-2 increased, and the protein level increased, but the difference was not statistically significant.