黄连有效成分盐酸小檗碱、盐酸药根碱、黄连碱及其混合物对NCI-H716细胞分泌胰高血糖素样肽1(GLP-1)的影响
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摘要
目的:研究黄连有效成分对NCI-H716细胞分泌胰高血糖素样肽1(GLP-1)的影响。方法:分别设置空白对照组、盐酸药根碱(JAT)组、盐酸小檗碱(BER)组、黄连碱(COP)组、盐酸巴马汀(BM)及BER十COP十JAT高剂量组和BER+COP+JAT低剂量组,同时以BER组为阳性对照组。采用CCK-8法检测各组药物不同浓度下对细胞增殖的抑制作用,确定药物浓度范围;采用酶联免疫吸附法(ELISA)检测NCI-H716细胞分泌GLP-1的量;采用聚合酶链式反应(PCR)检测NCI-H716细胞表达GLP-1RNA的量。结果:COP、JAT,以及高、低浓度BER+COP+JAT均能促进NCI-H716细胞分泌GLP-1(P<0.05),其中,BER+COP+JAT高浓度组及BER+COP+JAT低浓度组促GLP-1分泌作用均优于阳性对照组(P<0.05);COP、JAT组与阳性对照组相比无显著性差异(P>0.05)。结论:JAT、COP对NCI-H716细胞分泌GLP-1均有促进作用,且BER+COP+JAT混合组效果优于其单体组。
OBJECTIVE To investigate the effect of rhizoma coptidis on the expression of glucagon-like peptide 1(GLP-1).METHODS The control group,the jatrorrhizine hydrochloride(JAT)group,the coptisine(COP)group,the palmatine hydrochloride(BM)group,the berberine hydrochloride(BER)group,the low-dose combination group and the high-dose combination group were set,the BER group was set as the positive control group.CCK-8 assay was used to test the survival rate of NCI-H716 cells treated by JAT,COP,BM and BER.The expression of GLP-1 in NCI-H716 cells was detected by enzyme linked immunosorbent assay(ELISA),and the expression of RNA of GLP-1 in NCI-H716 cells was detected by polymerase chain reaction(PCR).RESULTS Secretion of GLP-1 in NCI-H716 cells was increased after exposure to JAT,COP and their combination(P<0.05),but not increased in the BM group(P>0.05).The GLP-1 secretion was highest in combination group at high dose and low dose.There was no significant difference between JAT,COP with the positive control group.CONCLUSION The COP and JAT can promote the secretion of GLP-1 in NCI-H716 cells,and the effects of combination group were better than the monomer group.
OBJECTIVE To investigate the effect of rhizoma coptidis on the expression of glucagon-like peptide 1(GLP-1).METHODS The control group,the jatrorrhizine hydrochloride(JAT)group,the coptisine(COP)group,the palmatine hydrochloride(BM)group,the berberine hydrochloride(BER)group,the low-dose combination group and the high-dose combination group were set,the BER group was set as the positive control group.CCK-8 assay was used to test the survival rate of NCI-H716 cells treated by JAT,COP,BM and BER.The expression of GLP-1 in NCI-H716 cells was detected by enzyme linked immunosorbent assay(ELISA),and the expression of RNA of GLP-1 in NCI-H716 cells was detected by polymerase chain reaction(PCR).RESULTS Secretion of GLP-1 in NCI-H716 cells was increased after exposure to JAT,COP and their combination(P<0.05),but not increased in the BM group(P>0.05).The GLP-1 secretion was highest in combination group at high dose and low dose.There was no significant difference between JAT,COP with the positive control group.CONCLUSION The COP and JAT can promote the secretion of GLP-1 in NCI-H716 cells,and the effects of combination group were better than the monomer group.
引文
[1] Cabou C,Burcelin R.GLP-1,the gut-brain,and brain-periphery axes[J].Rev Diabet Stud,2011,8(3):418-431.
[2] Wang L,Liu Y,Yang J,et al.GLP-1analogue liraglutide enhances proinsulin processing in pancreaticβ-cells via a PKA-dependent pathway[J].Endocrinology,2014,155(10):3817-3828.
[3] Song XY,He WJ,Sun XY,et al.Glucagon-Like Peptide-1and PancreaticβCells[J].Letters in Biotechnology,2015,26(4):579-583.
[4] Yu Y,Hao G,Zhang Q,et al.Berberine induces GLP-1secretion through activation of bitter taste receptor pathways[J].Biochemical Pharmacology,2015,97(2):173-177.
[5] Zhang Q,Xiao X,Li M,et al.Berberine moderates glucose metabolism through the GnRH-GLP-1and MAPK pathways in the intestine[J].BMC Complementary and Alternative Medicine,2014,14:188.
[6] Gupta R,Walunj SS,Tokala RK,et al.Emerging drug candidates of dipeptidyl peptidase IV(DPP IV)inhibitor class for the treatment of Type 2 Diabetes[J].Curr Drug Targets,2009,10(1):71-87.
[7] Zhao QP,Cui QB,Zhang Y.Research Progress of Coptis chinensis in treatment of Diabetes[J].Pharmacy and Clinics of Chinese Materia Medica(中药与临床),2010,1(1):55-58.
[8] Wei SC,Xu LJ,Zou X,et al.Pharmacokinetic and pharmacodynamic characteristics of berberine and jateorhizine in Coptidis Rhizoma powder and their monomeric compounds in type 2diabetic rats[J].China J Chin Mater Med(中国中药杂志),2015,40(21):4262-4267.
[9] Wei SC,Xu LJ,Zou X,et al.Preliminary study on secreting effects of Rhizoma Coptidis active ingredients and their combinations on insulin and glucagon-like peptide1(GLP-1)in glucose feeding mice[J].China J Chin Mater Med(中国中药杂志),2017,37(14):1343-1347.
[10]Zhu JY,Wu XH.Research Progress of Intestinal L cells[J].Chin J Diabetes Mellitus(中华糖尿病杂志),2012,4(1):51-53。
[11]Reimer RA,Darimont C,Gremlich S,et al.Ahumol·L-1an cellular model for studying the regulation of glucagon-like peptide-1secretion[J].Endocrinology,2001,142(10):4522-4528.
[12]Li WJ,Zheng J,LM,et al.The effects of Palmitic acid on the viability and enzymes involved in lipid metabolism of the intestinal endocrine GLP-1secreting cells.[A].Compilation of papers of the 12th national endocrinology academic conference of Chinese medical association[C].2013:223-224.
[13]Yu YL,Liu L,Wang XT,et al.Modulation of glucagon-like peptide-1release by berberine:In vivo and invitrostudies[J].Biochem Pharmacol,2010,79(7):1000-1006.
[14]Lu SS,Yu YL,Zhu HJ,et al.Berberine promotes glucagonlikepeptide-1(7-36)amide secretion in streptozotocin-induced diabetic rats[J].Endocrinol,2009,200(2):159-165.
[15]Kong WJ,Zhang H,Song DQ,et al.Berberine reduces insulin resistance through protein kinase C-dependent up-regulation of insulin receptor expression[J].Metabolism,2009,58(1):109-119.
[16]Dong H,Wang N,Zhao L,et al.Berberine in the treatment of type 2diabetes mellitus:a systemic review and meta-analysis[J].Evid Based Complement Alternat Med,2012:591654.
[17]Wang ZQ,Lu FE,Leng SH,et al.Facilitating effects of berberine on rat pancreatic islets through modulating hepatic nuclear factor 4alpha expression and glucokinase activity[J].World J Gastroenterol,2008,14(39):6004-6011.
[18]Lee YS,Kim WS,Kim KH,et al.Berberine,a natural plantproduct,activates AMP-activated protein kinase with beneficial metabolic effects in diabetic and insulin-resistant states[J].Diabetes,2006,55(8):2256-2264.
[19]Zhang Y,Li X,Zou D,et al.Treatment of type 2diabetes and dyslipidemia with the natural plant alkaloid berberine[J].Clin Endocrinol Metab,2008,93(7):2559-2565.
[20]Zhu SL,Lei T,Gao X,et al.Jatrorrhizine regulates GLUTs with multiple manners for hypoglycemic effect in insulin-resistance 3T3-L1adipocytes[J].China J Chin Mater Med(中国中药杂志),2018,43(6):1215-1220.
[21]Li DF,Yan J,Zhang KB,et al.Protective effect and mechanism of palmatine on diabetic mouse aorta[J].J Chongqing Med Univ(重庆医科大学学报),2017,42(11):1522-1526.
[22]Li C,He JZ,Zhou XD,et al.Berberine regulates type 2diabetes mellitus related with insulin resistance[J].China J Chin Mater Med(中国中药杂志),2017,42(12):2254-2260.
[23]Li Hua.The new power to treat type 2diabetes—GLP-1analogue liraglutide[J].Seek Medical And Ask The Medicine,2012,10(6):197-198.
[24]Zhao FQ,Xia Y,Zhao L,et al.Clinical Research Review of Semagutide[J].Heilongjiang Med J(黑龙江医药),2017,30(6):1218-1220.
[25]Meng YQ,Zhang Y,Liu FX,et al.Research progress in pharmacologic actions of dipeptidyl peptidase IV inhibitors and its mechanism[J].Drugs&Clinic,2013,28(2):101-107.
[26]Jiang Y,Peng YD.Research progress of human GLP-1analogues for type 2diabetes[J].World Clin Drugs(世界临床药物),2010,31(2):74-77.
[27]Li YL,Wang JX,Wu XM,et al.Recent progress in research and development of dipeptidyl peptidaseⅣinhibitors[J].Chin J New Drugs(中国新药),2008,17(20):1739-1745.
[2] Wang L,Liu Y,Yang J,et al.GLP-1analogue liraglutide enhances proinsulin processing in pancreaticβ-cells via a PKA-dependent pathway[J].Endocrinology,2014,155(10):3817-3828.
[3] Song XY,He WJ,Sun XY,et al.Glucagon-Like Peptide-1and PancreaticβCells[J].Letters in Biotechnology,2015,26(4):579-583.
[4] Yu Y,Hao G,Zhang Q,et al.Berberine induces GLP-1secretion through activation of bitter taste receptor pathways[J].Biochemical Pharmacology,2015,97(2):173-177.
[5] Zhang Q,Xiao X,Li M,et al.Berberine moderates glucose metabolism through the GnRH-GLP-1and MAPK pathways in the intestine[J].BMC Complementary and Alternative Medicine,2014,14:188.
[6] Gupta R,Walunj SS,Tokala RK,et al.Emerging drug candidates of dipeptidyl peptidase IV(DPP IV)inhibitor class for the treatment of Type 2 Diabetes[J].Curr Drug Targets,2009,10(1):71-87.
[7] Zhao QP,Cui QB,Zhang Y.Research Progress of Coptis chinensis in treatment of Diabetes[J].Pharmacy and Clinics of Chinese Materia Medica(中药与临床),2010,1(1):55-58.
[8] Wei SC,Xu LJ,Zou X,et al.Pharmacokinetic and pharmacodynamic characteristics of berberine and jateorhizine in Coptidis Rhizoma powder and their monomeric compounds in type 2diabetic rats[J].China J Chin Mater Med(中国中药杂志),2015,40(21):4262-4267.
[9] Wei SC,Xu LJ,Zou X,et al.Preliminary study on secreting effects of Rhizoma Coptidis active ingredients and their combinations on insulin and glucagon-like peptide1(GLP-1)in glucose feeding mice[J].China J Chin Mater Med(中国中药杂志),2017,37(14):1343-1347.
[10]Zhu JY,Wu XH.Research Progress of Intestinal L cells[J].Chin J Diabetes Mellitus(中华糖尿病杂志),2012,4(1):51-53。
[11]Reimer RA,Darimont C,Gremlich S,et al.Ahumol·L-1an cellular model for studying the regulation of glucagon-like peptide-1secretion[J].Endocrinology,2001,142(10):4522-4528.
[12]Li WJ,Zheng J,LM,et al.The effects of Palmitic acid on the viability and enzymes involved in lipid metabolism of the intestinal endocrine GLP-1secreting cells.[A].Compilation of papers of the 12th national endocrinology academic conference of Chinese medical association[C].2013:223-224.
[13]Yu YL,Liu L,Wang XT,et al.Modulation of glucagon-like peptide-1release by berberine:In vivo and invitrostudies[J].Biochem Pharmacol,2010,79(7):1000-1006.
[14]Lu SS,Yu YL,Zhu HJ,et al.Berberine promotes glucagonlikepeptide-1(7-36)amide secretion in streptozotocin-induced diabetic rats[J].Endocrinol,2009,200(2):159-165.
[15]Kong WJ,Zhang H,Song DQ,et al.Berberine reduces insulin resistance through protein kinase C-dependent up-regulation of insulin receptor expression[J].Metabolism,2009,58(1):109-119.
[16]Dong H,Wang N,Zhao L,et al.Berberine in the treatment of type 2diabetes mellitus:a systemic review and meta-analysis[J].Evid Based Complement Alternat Med,2012:591654.
[17]Wang ZQ,Lu FE,Leng SH,et al.Facilitating effects of berberine on rat pancreatic islets through modulating hepatic nuclear factor 4alpha expression and glucokinase activity[J].World J Gastroenterol,2008,14(39):6004-6011.
[18]Lee YS,Kim WS,Kim KH,et al.Berberine,a natural plantproduct,activates AMP-activated protein kinase with beneficial metabolic effects in diabetic and insulin-resistant states[J].Diabetes,2006,55(8):2256-2264.
[19]Zhang Y,Li X,Zou D,et al.Treatment of type 2diabetes and dyslipidemia with the natural plant alkaloid berberine[J].Clin Endocrinol Metab,2008,93(7):2559-2565.
[20]Zhu SL,Lei T,Gao X,et al.Jatrorrhizine regulates GLUTs with multiple manners for hypoglycemic effect in insulin-resistance 3T3-L1adipocytes[J].China J Chin Mater Med(中国中药杂志),2018,43(6):1215-1220.
[21]Li DF,Yan J,Zhang KB,et al.Protective effect and mechanism of palmatine on diabetic mouse aorta[J].J Chongqing Med Univ(重庆医科大学学报),2017,42(11):1522-1526.
[22]Li C,He JZ,Zhou XD,et al.Berberine regulates type 2diabetes mellitus related with insulin resistance[J].China J Chin Mater Med(中国中药杂志),2017,42(12):2254-2260.
[23]Li Hua.The new power to treat type 2diabetes—GLP-1analogue liraglutide[J].Seek Medical And Ask The Medicine,2012,10(6):197-198.
[24]Zhao FQ,Xia Y,Zhao L,et al.Clinical Research Review of Semagutide[J].Heilongjiang Med J(黑龙江医药),2017,30(6):1218-1220.
[25]Meng YQ,Zhang Y,Liu FX,et al.Research progress in pharmacologic actions of dipeptidyl peptidase IV inhibitors and its mechanism[J].Drugs&Clinic,2013,28(2):101-107.
[26]Jiang Y,Peng YD.Research progress of human GLP-1analogues for type 2diabetes[J].World Clin Drugs(世界临床药物),2010,31(2):74-77.
[27]Li YL,Wang JX,Wu XM,et al.Recent progress in research and development of dipeptidyl peptidaseⅣinhibitors[J].Chin J New Drugs(中国新药),2008,17(20):1739-1745.