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时间:2020-05-18
《脱氢紫堇碱对高糖培养心肌成纤维细胞增殖和胶原分泌的影响.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、·5O·ChineseJournaIofInformationonTCMOct.2014VoI.21No.1O脱氢紫堇碱对高糖培养心肌成纤维细胞增殖和胶原分泌的影响韩路拓’,杨佳妹’,任钧国,李鸿海,李军梅,刘建勋1.北京中医药大学,北京100029;2.中国中医科学院西苑医院,中药药理北京市重点实验室,北京100091摘要:目的观察中药延胡索主要有效成分脱氢紫堇碱(DHC)对高浓度葡萄糖培养的心肌成纤维细胞(CFs)增殖和胶原分泌的影响,为延胡索的临床应用提供实验依据。方法采用体外高糖培养的CFs模型进行实验。以胶原
2、酶与胰蛋白酶联合消化分离出生24h内乳鼠CFs,取2~4代细胞高浓度葡萄糖(25mmol/L)培养,分别加入100、50、25mg/LDHC干预,24、48h后镜下观察细胞形态,MTT法检测细胞增殖,流式细胞仪检测细胞周期,ELISA测定I型、III型胶原含量。结果CFs种植3h后开始贴壁生长,高浓度葡萄糖培养液培养24、48h后细胞增殖明显增加(P3、殖(P4、rationandCollagenSecretionofCardiacFibroblastsCulturedbyHighGlucoseHANLu.tuo,,YANGJia.mei'。,RENJun.guo,LIHonghai,LIJun—mei,LIUJian-xunf1.BeijingUniversityofChineseMedicine.Beijing100029,China:2.XiyuanHospitalMedicalofChinaAcademyofChineseSciences,KeyLaboratoryof5、PharmacologyofChineseMateriainBeijing,Beijing100091,China)Abstract:ObjectiveToobservetheeffectsofdehydrocorydaline(DHC)onproliferationandcollagensecretionofcardiacfibroblasts(CFs)culturedbyhighglucose:ToprovideexperimentalevidenceforclinicalapplicationofRhizomaC6、orydalis.MethodsCFsculturedinvitrowithhighglucoseweremadeintomodels.CollagenaseandtrypsinwereusedforthecombinedigestionofCFsfromratsbornin24h.2-4generationCFswereculturedbyhighglucose(25mmol/L】,andthen100,50,25mg/Ldethydrocorydalinewasaddedforintervention.Cellul7、armorphologyofCFswasobservedafter24,48h.CFsproliferationwasdetectedbyMTTmethod.Cellcyclewasassessedviaflowcytometry.ThelevelsofcollagenIandcollagenII1weredeterminedbyELISA.ResultsCFsbegantogrowadherence3hoursafterplanting,andCFsculturedbyhighglucosesignificantly8、proliferated24,48hlater(P
3、殖(P4、rationandCollagenSecretionofCardiacFibroblastsCulturedbyHighGlucoseHANLu.tuo,,YANGJia.mei'。,RENJun.guo,LIHonghai,LIJun—mei,LIUJian-xunf1.BeijingUniversityofChineseMedicine.Beijing100029,China:2.XiyuanHospitalMedicalofChinaAcademyofChineseSciences,KeyLaboratoryof5、PharmacologyofChineseMateriainBeijing,Beijing100091,China)Abstract:ObjectiveToobservetheeffectsofdehydrocorydaline(DHC)onproliferationandcollagensecretionofcardiacfibroblasts(CFs)culturedbyhighglucose:ToprovideexperimentalevidenceforclinicalapplicationofRhizomaC6、orydalis.MethodsCFsculturedinvitrowithhighglucoseweremadeintomodels.CollagenaseandtrypsinwereusedforthecombinedigestionofCFsfromratsbornin24h.2-4generationCFswereculturedbyhighglucose(25mmol/L】,andthen100,50,25mg/Ldethydrocorydalinewasaddedforintervention.Cellul7、armorphologyofCFswasobservedafter24,48h.CFsproliferationwasdetectedbyMTTmethod.Cellcyclewasassessedviaflowcytometry.ThelevelsofcollagenIandcollagenII1weredeterminedbyELISA.ResultsCFsbegantogrowadherence3hoursafterplanting,andCFsculturedbyhighglucosesignificantly8、proliferated24,48hlater(P
4、rationandCollagenSecretionofCardiacFibroblastsCulturedbyHighGlucoseHANLu.tuo,,YANGJia.mei'。,RENJun.guo,LIHonghai,LIJun—mei,LIUJian-xunf1.BeijingUniversityofChineseMedicine.Beijing100029,China:2.XiyuanHospitalMedicalofChinaAcademyofChineseSciences,KeyLaboratoryof
5、PharmacologyofChineseMateriainBeijing,Beijing100091,China)Abstract:ObjectiveToobservetheeffectsofdehydrocorydaline(DHC)onproliferationandcollagensecretionofcardiacfibroblasts(CFs)culturedbyhighglucose:ToprovideexperimentalevidenceforclinicalapplicationofRhizomaC
6、orydalis.MethodsCFsculturedinvitrowithhighglucoseweremadeintomodels.CollagenaseandtrypsinwereusedforthecombinedigestionofCFsfromratsbornin24h.2-4generationCFswereculturedbyhighglucose(25mmol/L】,andthen100,50,25mg/Ldethydrocorydalinewasaddedforintervention.Cellul
7、armorphologyofCFswasobservedafter24,48h.CFsproliferationwasdetectedbyMTTmethod.Cellcyclewasassessedviaflowcytometry.ThelevelsofcollagenIandcollagenII1weredeterminedbyELISA.ResultsCFsbegantogrowadherence3hoursafterplanting,andCFsculturedbyhighglucosesignificantly
8、proliferated24,48hlater(P
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