Initial Study About The PLCs On The Insulin Secretion

CHAPTER1 EFFECTS OF PHOSPHOLIPASE C IN THE INSULIN SECRETIONObjective: To investigate the role of PLC in insulin secretion after the stimulation of nutrient substance,ion,neurotransmitter,hormone.Methods :①Designed the five stimulating factors:glucose,L-aminoglutaminic acid,KCL,cholestokinin-octopeptide, acetylcholine chloride.②Setting the concentration group and the time group, stimulated INS-1 respectively③Using the rat insulin ELISA kit to detect the content of insulin in INS-1's supernatant liquid.④RT-PCR was used to detect the expression of PLC.Results:①PLCεhad no expression in INS-1;②RT-PCR was used to detect the change quantity of expression of PLC;the group of glucose: PLCδ> PLCβ> PLCγ> PLCη> PLCζ,the group of L-aminoglutaminic acid: PLCβ> PLCδ> PLCζ> PLCη>PLCγ,the group of KCL: PLCη> PLCδ> PLCγ> PLCζ>PLCβ,the group of cholestokinin-octopeptide:PLCδ> PLCγ> PLCη> PLCζ> PLCβ,the group of acetylcholine chloride: PLCδ> PLCβ> PLCγ> PLCη> PLCζ Conclusion:The expression level of PLC up-regulated significantly conspicuously after the effects of glucose,L-aminoglutaminic acid,KCL,cholestokinin-octopeptide, acetylcholine chloride,but the change quantity were distinct.CHAPTER 2 EFFECTS OF OVEREXPRESSION OF PHOSPHOLIPASE Cβ1 ONβCELL GLUCOSE-STIMULATED INSULIN SECRETIONObjective:To investigate the role of PLCβ1 in glucose-stimulated insulin secretion(GSIS).Methods:①RT-PCR was used to detect the expression of PLCβ1 .②Eukaryotic expression vector PCMV-HA-PLCβ1 was constrcted and then transient transfected into INS-1. The PLCβ1 protein was detected by western blot.③Using the rat insulin ELISA kit to detect the content of insulin in INS-1's supernatant liquid.Results : The expression of PLCβ1 was raised by glucose's stimulating.The insulin content in supernatant liquid of INS-1,in which PLCβ1 was overexpressed,was raised(1.906±0.080) ng/ml ( p<0.01)compared with the control( 0.740±0.091 ) ng/ml.Conclusion:Transient overexpression of PLCβ1 in INS-1 significantly increased insulin secretion, which suggested that PLCβ1 may be involved in the signal transduction pathway of GSIS in INS-1cells.