The Mechanism Of Qingrejiangzhuo Decotion In Promoting Glucagon-like Peptide1Secretion Through Regulating The Expression Of G-protein Coupled Receptor119

Part1The effection of Qingrejiangzhuo Decotion and berberine on the proliferation and apoptosis of IEC-6cells,and the effects of Qingrejiangzhuo Decotion and berberine on GLP-1and cAMP secretion of IEC-6cellsObjective:To observe the effection of Qingrejiangzhuo Decotion and berberine on the proliferation and apoptosis of IEC-6cells,and the effects of Qingrejiangzhuo Decotion and berberine on GLP-1secretion of IEC-6cells,and whether the effects is related to the activation of GPR119.Methods:The proliferation of IEC-6cells was observed by MTT assay.The apoptosis of IEC-6cells was observed through by flow cytometry.The content of GLP-1,GPR119and cAMP was determined by radioimmunoassay.Results:The survival rate of IEC-6cells was significantly raised after intervention with Qingrejiangzhuo Decotion,comparing with normal culture group. The survival rate of IEC-6cells was highest after intervention with2.5%medicated serum,with a survival rate of162.42%.So2.5%medicated serum was chosed as the working concentration. The survival rate of IEC-6cells was significantly raised after intervention with berberine,comparing with normal culture group. The survival rate of IEC-6cells was highest after intervention with1μg/mLberberine,with a survival rate of220.17%.So1μg/mLberberine solution was chosed as the working concentration.The apotosis rate of IEC-6cells decreased significantly after Qingrejiangzhuo Decotion and berberine intervention.Compared with the normal culture group,the apoptosis rate of DMEM high glucose medium containing medicated serum group decreased by7.92%,the apoptosis rate of DMEM high glucose medium containing berberine group decreased by6.23%.Compared with the DMEM low glucose medium containing fetal bovine serum group,the apoptosis rate of DMEM low glucose medium containing medicated serum group decreased by10.92%,the apoptosis rate of DMEM low glucose medium containing berberine group decreased by8.51%.Compared with the DMEM higer glucose medium containing fetal bovine serum group,the apoptosis rate of DMEM higher glucose medium containing berberine group decreased by9.3%.The content of GLP-1of the DMEM high glucose medium containing medicated serum group was2.167±0.167pmol/g, the DMEM high glucose medium containing berberine group1.870±0.098pmol/g,both significantly higher than the normal culture group(1.687±0.196pmol/g,P<0.01).The content of GLP-1of the DMEM low glucose medium containing medicated serum group was1.942±0.119pmol/g,the DMEM low glucose medium containing berberine group1.565±0.020pmol/g,both significantly higher than the DMEM low glucose medium containing fetal bovine serum group (1.441±0.102pmol/g,P<0.01).The content of GLP-1of the DMEM higher glucose medium containing berberine group2.286±0.105pmol/g, significantly higher than the DMEM higher glucose medium containing fetal bovine serum group (1.856±0.015,P<0.01).The content of cAMP of the DMEM high glucose medium containing medicated serum group was1.820±0.128pmol/mg,the DMEM high glucose medium containing berberine group1.683±0.063pmol/mg,both significantly higher than the normal culture group (1.539±0.065pmol/mg,P<0.01).The content of cAMP of the DMEM low glucose medium containing medicated serum group was1.457±0.057pmol/mg,the DMEM low glucose medium containing berberine group1.428±0.053pmol/mg,both significantly higher than the DMEM low glucose medium containing fetal bovine serum group (1.249±0.046pmol/mg.P<0.01).The content of cAMP of the DMEM higher glucose medium containing berberine group1.799±0.045pmol/g,significantly higher than the DMEM higher glucose medium containing fetal bovine serum group (1.655±0.069,P<0.01)Conclusion:Qingrejiangzhuo decotion could promote the proliferation of IEC-6cells,reduce the apoptosis of IEC-6cells,and promote IEC-6cells to secrete GLP-1.The promotion effects could be attributed to the activation of GPR119-cAMP-GLP-1pathway.The berberine showed the same effects with Qingrejiangzhuo decotion. Part2The effection of Qingrejiangzhuo Decotion and berberine on the expression of protein and mRNA of GLP-1and GPR119in IEC-6cellsObjective:To observe the promotion effects of Qingrejiangzhuo decotion and berberine on GLP-1secretion whether can be attributed to the promotion effects on the expression of GPPR119.Methods:The relative expression of mRNA of GLP-1and GPR119were determined by RT-PCR,calculating2(-Delta Delta C[T]).The relative expression of proteins of GLP-1and GPR119were determined by Western blot.Results:The relative expression of GLP-1mRNA of the DMEM high glucose medium containing medicated serum group was4.90359±0.273842,the relative expression of GLP-1mRNA of the DMEM high glucose medium containing berberine group was4.75654±0.244486,the relative expression of GLP-1mRNA of the DMEM higher glucose medium containing berberine group was4.95339±0.216146,the relative expression of GLP-1mRNA of the DMEM higher glucose medium containing fetal bovine serum group was4.47110±0.199938,the relative expression of GLP-1mRNA of the DMEM low glucose medium containing medicated serum group was1.04591±0.163524,the relative expression of GLP-1mRNA of the DMEM low glucose medium containing berberine group was0.94792±0.085629,and the relative expression of GLP-1mRNA of the DMEM low glucose medium containing fetal bovine serum group was0.54016±0.041613.The relative expressions of GLP-1mRNA of the DMEM high glucose medium containing medicated serum group and the DMEM high glucose medium containing berberine group were significantly higher than the normal culture group (P<0.01).The relative expressions of GLP-1mRNA of the DMEM low glucose medium containing medicated serum group and the DMEM low glucose medium containing berberine group were significantly higher than the DMEM low glucose medium containing fetal bovine serum group (P<0.01).The relative expression of GLP-1mRNA of the DMEM higher glucose medium containing berberine group was higher than the DMEM higher glucose medium containing fetal bovine serum group (P<0.01).The relative expression of GPR119mRNA of the DMEM high glucose medium containing medicated serum group was5.00823±0.294559,the relative expression of GPR119mRNA of the DMEM high glucose medium containing berberine group was4.75283±0.307996,the relative expression of GPR119mRNA of the DMEM higher glucose medium containing berberine group was4.91934±0.284853,the relative expression of GPR119mRNA of the DMEM higher glucose medium containing fetal bovine serum group was4.35557±0.237960,the relative expression of GPR119mRNA of the DMEM low glucose medium containing medicated serum group was1.00210±0.107958,the relative expression of GPR119mRNA of the DMEM low glucose medium containing berberine group was0.95795±0.075175,and the relative expression of GPR119mRNA of the DMEM low glucose medium containing fetal bovine serum group was0.44108±0.034275.The relative expressions of GPR119mRNA of the DMEM high glucose medium containing medicated serum group and the DMEM high glucose medium containing berberine group were significantly higher than the normal culture group (P<0.01).The relative expressions of GPR119mRNA of the DMEM low glucose medium containing medicated serum group and the DMEM low glucose medium containing berberine group were significantly higher than the DMEM low glucose medium containing fetal bovine serum group(P<0.01).The relative expression of GPR119mRNA of the DMEM higher glucose medium containing berberine group was higher than the DMEM higher glucose medium containing fetal bovine serum group (P<0.01).The relative expression of protein of GLP-1was1.66333±0.074976of the DMEM high glucose medium containing medicated serum group,the relative expression of protein of GLP-1was1.26583±0.025262of the DMEM high glucose medium containing berberine group,the relative expression of protein of GLP-1was0.90643±0.025290of the DMEM high glucose medium containing fetal bovine serum group,the relative expression of protein of GLP-1was1.63833±0.052504of the DMEM higher glucose medium containing berberine group,the relative expression of protein of GLP-1was1.24667±0.035387of the DMEM higher glucose medium containing fetal bovine serum group,the relative expression of protein of GLP-1was0.80650±0.024825of the DMEM low glucose medium containing medicated serum group,the relative expression of protein of GLP-1was0.79833±0.025201of the DMEM low glucose medium containing berberine group,and the relative expression of protein of GLP-1was0.49217±0.043582of the DMEM low glucose medium containing fetal bovine serum group.The protein relative expressions of GLP-1of the DMEM high glucose medium containing medicated serum group and the DMEM high glucose medium containing berberine group were significantly higher than the normal culture group (P<0.01). The protein relative expressions of GLP-1of the DMEM low glucose medium containing medicated serum group and the DMEM low glucose medium containing berberine group were significantly higher than the DMEM low glucose medium containing fetal bovine serum group (P<0.01).The protein relative expression of GLP-1of the DMEM higher glucose medium containing berberine group was higher than the DMEM higher glucose medium containing fetal bovine serum group (P<0.01). The relative expression of protein of GPR119was1.59017±0.056912of the DMEM high glucose medium containing medicated serum group,the relative expression of protein of GPR119was1.37133±0.035478of the DMEM high glucose medium containing berberine group,the relative expression of protein of GPR119was0.77183±0.025725of the DMEM high glucose medium containing fetal bovine serum group,the relative expression of protein of GPR119was1.60883±0.057881of the DMEM higher glucose medium containing berberine group,the relative expression of protein of GPR119was0.98058±0.062956of the DMEM higher glucose medium containing fetal bovine serum group,the relative expression of protein of GPR119was0.67033±0.040820of the DMEM low glucose medium containing medicated serum group,the relative expression of protein of GPR119was0.66643±0.022541of the DMEM low glucose medium containing berberine group,and the relative expression of protein of GPR119was0.45283±0.024879of the DMEM low glucose medium containing fetal bovine serum group.The protein relative expressions of GPR119of the DMEM high glucose medium containing medicated serum group and the DMEM high glucose medium containing berberine group were significantly higher than the normal culture group(P<0.01). The protein relative expressions of GPR119of the DMEM low glucose medium containing medicated serum group and the DMEM low glucose medium containing berberine group were significantly higher than the DMEM low glucose medium containing fetal bovine serum group (P<0.01). The protein relative expression of GPR119of the DMEM higher glucose medium containing berberine group was higher than the DMEM higher glucose medium containing fetal bovine serum group (P<0.01)Conclusion:Qingrejiangzhuo decotion and berberine can promote the expression of GLP-1in IEC-6cells,and the mechanism of promotion can be attributed to the up-regulation of GPR119expression.