Effects Of MiR-15b-5p On Apoptosis Of Rat Pancreatic Exocrine Cells AR42J

Objective Acute pancreatitis（AP）is a gastrointestinal disease with pancreatic acinar cell injury and apoptosis,pancreatic inflammation and strong systemic inflammatory response.The regulation of apoptosis plays an important role in the growth and development of the whole organism and the development of diseases.mi R-15b-5p plays a role in regulating cell apoptosis and affecting disease development in many diseases.The role of mi R-15b-5p in apoptosis in acute pancreatitis has not been studied,so this paper aims to explore whether mi R-15b-5p is involved in the regulation of apoptosis in acute pancreatitis.Methods AR42 J cells were cultured to the logarithmic growth phase in vitro,and the AP model was established by adding 100 n M cerulein.After the model was completed,mi RNA was extracted and q RT-PCR was used to detect the relative expression of mi R-15b-5p.Subsequently,mi R-15b-5p mimics,mi R-15b-5p mimics NC,mi R-15b-5p inhibitor,and mi R-15b-5p inhibitor NC were transfected into AR42 J cells respectively with transfection reagents.After 24 h of transfection,mi RNA was extracted to detect the relative expression of mi R-15b-5p by q RT-PCR.Then,the cells in the logarithmic growth phase were seeded in a six-well plate,and the cells were transfected according to the above groups.After 24 h,the cells in each well were treated with caerulein for 24 h to construct an in vitro acute pancreatitis model.The relative expression of mi R-15b-5p was detected by q RT-PCR,and the relative expression levels of Caspase-3 and Caspase-9 m RNA were detected by q RT-PCR.The cell proteins were extracted and the relative expression levels of Caspase-3 and Caspase-9 proteins were determined by Western blot.Flow cytometry was used to determine the apoptosis of cells in each group.Results（1）In AP model,q RT-PCR results showed that the relative expression of mi R-15b-5p in AP group was higher than that in blank control group（P < 0.05）.（2）After transfection,the relative expression level of mi R-15b-5p increased in mi R-15b-5p mimics group（P < 0.05）,and decreased in mi R-15b-5p inhibitor group（P < 0.05）,indicating that the transfection was successful.（3）After using mi R-15b-5p inhibitor,the m RNA relative expression levels of apoptosis-related genes Caspase-3 and Caspase-9 decreased（P <0.05）,while after using mimics,the m RNA relative expression level increased（P <0.05）.（4）Western blot showed that the relative expression levels of Caspase-3 and Caspase-9 proteins related to apoptosis were decreased after mi R-15b-5p inhibitor was used（P <0.05）,while the relative expression levels of proteins were increased after mimics was used（P <0.05）.（5）Flow cytometry showed that the incidence of cell death decreased after the use of mi R-15b-5p inhibitor（P <0.05）,while the incidence of apoptosis increased after the use of mimics（P <0.05）.Conclusion When pancreatitis occurs,the expression of mi R-15b-5p is relatively higher,mi R-15b-5p inhibitor can reduce the apoptosis of pancreatic exocrine cells induced by cerulein,and increase apoptosis after using mi R-15b-5p mimics.mi R-15b-5p may activate downstream Caspase-3 by activating Caspase-9 and induce apoptosis.It may be an effective potential target for the prevention and treatment of acute pancreatitis.