The Effect Of SAP Interrelated MiR-551b-5p To The C-KIT Gene Expression In ICC And The Intracellular Concentration Of Calcium Ion

Objective: To discuss the mechanism by which SAP-relevant-miR-551b-5p effects the gastrointestinal motility by researching the effect of severe acute pancreatitis(SAP)-relevant-miR-551b-5p on the c-kit gene expression and calcium concentration in Interstitial cells of Cajal(ICC).Methods: 1?The ICC was isolated and cultured. Small intestine was obtained from the SD neonatal rat. The mesentery, blood vessels and serosal layer were stripped under the microscope. Further more, the mucosa and submucosa of small intestine were stripped and ICC was obtained from the smooth muscle layer and cultured. 2?The cultured ICC was divided into five groups according to the research objectives: A: Normal comparison group.B:Mi R-551b-5p overexpression group(Electransfection the Mi R-551b-5p mimic).C: Mi R-551b-5p mimic negative control group. D: Mi R-551b-5p down-regulated expression group(Electransfection the Mi R-551b-5p inhibitor). E: Mi R-551b-5p inhibitor negative control group. 3?Testing the specificity Mi R-551b-5p expression in ICC which raised and lowered it expression or before it expressed. a?The Mi R-551b-5p and c-KIT m RNA expression in ICC was tested using RT-PCR technique and was repeated hree times, according to the sectionalization. b?The c-KIT protein expression in ICC was tested using Western blotting technique and was repeated three times. c?The morphologic change of ICC and the concentration change of calcium ion in ICC were tested using the Laser Scanning Confocal Microscope Testing. The expression of c-KIT gene was tested using the Immunofluorescence technique and the concentration change of calcium ion was tested using the Fluo-3AM technique.Result: 1?Compared with the normal comparison group,the miR-551b-5p expression was increased remarkably in miR-551b-5p mimic group(P<0.01);the miR-551b-5p expression was decreased remarkably in miR-551b-5p inhibitor group(P<0.01);Compared with the Mi R-551b-5p mimic negative control group, the miR-551b-5p expression was increased remarkably in miR-551b-5p mimic group(P<0.01);Compared with the Mi R-551b-5p inhibitor negative control group, the miR-551b-5p expression was decreased remarkably in miR-551b-5p inhibitor group. 2 ? Compared with the normal comparison group, there are no obvious differences in the expression of c-KIT gene in both miR-551b-5p mimic group and miR-551b-5p inhibitor group(P>0.05). 3?Compared with the normal comparison group,the calcium ion concentration of ICC increased remarkably in miR-551b-5p mimic group(P<0.05);the calcium ion concentration of ICC decreased remarkably in miR-551b-5p inhibitor group(P<0.05).Conclusion: 1?The expression of SAP–relavant-miR-551b-5p has positive correlation with the calcium ion concentration of ICC, by the means of the calcium concentration increasing, the ICC Paces malfunction, then it lead to the GID. 2?The expression of SAP relevancy miR-551b-5p has no effect on the c-KIT gene expressing in ICC.