欢迎来到天天文库
浏览记录
ID:28570261
大小:11.09 MB
页数:52页
时间:2018-12-11
《mapk在葛根素白藜芦醇促成骨细胞增殖分化和抑凋亡中的作用》由会员上传分享,免费在线阅读,更多相关内容在应用文档-天天文库。
1、p38MAPK在葛根素、白藜芦醇促成骨细胞增殖、分化和抑凋亡中的作用摘要目的探讨p38MAPK信号转导通路在178.雌二醇(17B.E2)、葛根素(Ptm)和白藜芦醇(RES)促成骨细胞(OB)增殖、分化、抑凋亡中的作用。方法1.用酶消化法分离培养新生大鼠颅骨成骨细胞,观察细胞形态,碱性磷酸酶(创LP)染色和矿化结节染色鉴定OB。2.采用第二代细胞,接种24h后分为对照组(Contr01);不同浓度(1ar8.10击tool·L-1)的178.E2、PUE和RES组;含阻断剂组178.E2+SB、PUE+SB和RES+SB,先加入SB203580(10,umol·L-1)作
2、用30分钟预先阻断p38MAPK信号转导通路,再加入上述浓度的178.E2、PUE和RES,培养72h后用M'IT法测定细胞的增殖能力,PNPP法检测ALP活性;茜素红染色计数矿化结节,测定钙(Ca)沉积量;反映OB矿化能力。3.细胞培养8d后采用RT.PCR半定量法测定10-Smol·L.1含与不含阻断剂的175.E2、PUE和RES组及对照组的I型胶原(CoilI)mRNA表达,反映向OB分化状况。4.第二代OB,接种24h后换无血清培养并分为对照组,10‘8tool·L-1含与不含阻断剂的17B.E2、PUE和RES组,培养72h后在AnnexinV.FITC/PI双
3、标记流式细胞仪上分析细胞早期凋亡。结果1.与对照组相比,17B.E2、PUE和RES各浓度MTI"值均显著增高伊4、UE+SB和RES+SB各组矿化结节数目及钙含量显著下降(P5、PUE和RES促OB的分化、矿化和I型胶原的表达过程中发挥重要作用,而对其促OB的增殖和抑制凋亡的作用无明显影响。关键词p38MAPKt成骨细胞;增殖;分化;凋亡;1713.雌二醇;葛根素;白藜芦醇UEffectsofP38ML廿KontheProliferation、DifferentiationandInhibitedApoptosisofMurineOsteoblastsbyPuerainandResveratr01.AbstractObjectiveToinvestigatetheeffectsofp38MAPKontheproliferation、differen6、tiationandinhibitedapoptosisofmurineostcoblastsby17B—estradiol,puerainandresveratr01.Methods1.ThenewbornSDmicecalvariumosteoblasts(OB)wereisolatedanddigestedwithtrypsinandcollagenaserepeatedly,whichwerethenculturedandobservedthemorphologiccharacterbothinthelightandphase-contrastmicroscopy.7、OBWasidentifiedbycytochemicalstainingofalkalinephosphatase(ALP)andthemineralizationnodeswerestainedbydifferentstaining.2.ThesecondpassagaeofOBwereplacein96-wellcultureplatesandincubatedfor24h,dividedtocontrol,variousconcentrationsof17B·estradiol,puerarinandres
4、UE+SB和RES+SB各组矿化结节数目及钙含量显著下降(P5、PUE和RES促OB的分化、矿化和I型胶原的表达过程中发挥重要作用,而对其促OB的增殖和抑制凋亡的作用无明显影响。关键词p38MAPKt成骨细胞;增殖;分化;凋亡;1713.雌二醇;葛根素;白藜芦醇UEffectsofP38ML廿KontheProliferation、DifferentiationandInhibitedApoptosisofMurineOsteoblastsbyPuerainandResveratr01.AbstractObjectiveToinvestigatetheeffectsofp38MAPKontheproliferation、differen6、tiationandinhibitedapoptosisofmurineostcoblastsby17B—estradiol,puerainandresveratr01.Methods1.ThenewbornSDmicecalvariumosteoblasts(OB)wereisolatedanddigestedwithtrypsinandcollagenaserepeatedly,whichwerethenculturedandobservedthemorphologiccharacterbothinthelightandphase-contrastmicroscopy.7、OBWasidentifiedbycytochemicalstainingofalkalinephosphatase(ALP)andthemineralizationnodeswerestainedbydifferentstaining.2.ThesecondpassagaeofOBwereplacein96-wellcultureplatesandincubatedfor24h,dividedtocontrol,variousconcentrationsof17B·estradiol,puerarinandres
5、PUE和RES促OB的分化、矿化和I型胶原的表达过程中发挥重要作用,而对其促OB的增殖和抑制凋亡的作用无明显影响。关键词p38MAPKt成骨细胞;增殖;分化;凋亡;1713.雌二醇;葛根素;白藜芦醇UEffectsofP38ML廿KontheProliferation、DifferentiationandInhibitedApoptosisofMurineOsteoblastsbyPuerainandResveratr01.AbstractObjectiveToinvestigatetheeffectsofp38MAPKontheproliferation、differen
6、tiationandinhibitedapoptosisofmurineostcoblastsby17B—estradiol,puerainandresveratr01.Methods1.ThenewbornSDmicecalvariumosteoblasts(OB)wereisolatedanddigestedwithtrypsinandcollagenaserepeatedly,whichwerethenculturedandobservedthemorphologiccharacterbothinthelightandphase-contrastmicroscopy.
7、OBWasidentifiedbycytochemicalstainingofalkalinephosphatase(ALP)andthemineralizationnodeswerestainedbydifferentstaining.2.ThesecondpassagaeofOBwereplacein96-wellcultureplatesandincubatedfor24h,dividedtocontrol,variousconcentrationsof17B·estradiol,puerarinandres
此文档下载收益归作者所有