中药筋脉通对高糖培养海马神经元细胞BAX, BCL3 蛋白.docVIP

中国康复理论与实践2012年4月第18卷第4期ChinJRehabilTheoryPractApr.2012Vol.18No.4DOI:10.3969/j.issn.1006-9771.2012.04.003·基础研究·中药筋脉通对高糖培养海马神经元细胞Bax、Bcl-2和Caspase-3蛋白表达的影响郭蕾蕾1a张宏1b田国庆1a梁晓春1a摘要目的探讨中药筋脉通含药血清对高糖培养海马神经元细胞的保护作用。方法采用血清药理学方法制备筋脉通含药血清JMT原代培养新生鼠海马神经元细胞分别接种于不同条件的培养基中分为正常对照组、高糖组、阳性对照组以及JMT大、中、小剂量组。不同培养基中培养72h后使用Westernblotting检测其Bax、Bcl-2和Caspase-3蛋白的表达。结果与正常对照组相比高糖组Caspase-3、Bax表达明显增加Plt0.01Bcl-2的表达下降Plt0.05与高糖组相比阳性对照组和JMT各剂量组的Caspase-3、Bax表达明显下降Plt0.01Bcl-2的表达升高Plt0.05与阳性对照组相比JMT各剂量组的Caspase-3、Bax表达下降Plt0.05筋脉通大、中剂量组Bcl-2的表达增加Plt0.05。结论筋脉通含药血清可能通过提高Bcl-2蛋白的表达减少Bax、Caspase-3的表达上调Bcl-2/Bax的比值从而减少海马神经元细胞的凋亡。关键词糖尿病认知功能障碍筋脉通海马神经元BaxBcl-2Caspase-3EffectsofJinmaitongonExpressionofBaxBcl-2andCaspase-3inHippocampalNeuronsCulturedwithHighGlucoseGUOLei-leiZHANGHongTIANGuo-qingetal.PekingUnionMedicalCollegeHospitalandInstituteofBasicMedicalSciencesPekingUnionMedicalCollegeChineseAcademyofMedicalSciencesBeijing100730ChinaAbstract:ObjectiveToexploretheeffectofJinmaitongJMTonexpressionofBaxBcl-2andCaspase-3inhippocampalneuronscul-turedwithhighglucose.MethodsHippocampalneuronswereprimarilyculturedandpurifiedfromthehippocampusofnew-bornSpragueDawleyrats.Neuron-specificenolaseimmunocytochemicalmethodwasadoptedfortheidentificationoftheneurons.Theyweredividedin-tonormalcontrolgrouphighglucosegrouphigh-doseJMTJHgroupmiddle-doseJMTJMgrouplow-doseJMTJLgroupandaposi-tivecontrolgroup.72hourslaterWesternblottingwasadoptedtodetecttheexpressionofBaxBcl-2andCaspase-3.ResultsComparedwiththenormalcontrolgrouptheexpressionofBaxandCaspase-3inthehighglucosegroupsignificantlyincreasedPlt0.01andtheex-pressionofBcl-2significantlydecreasedPlt0.05.ComparedwiththehighglucosegrouptheexpressionofCaspase-3andBaxintheposi-tivecontrolgroupandJHJMandJLgroupssignificantlydecreasedPlt0.01andtheexpressionofBcl-2significantlyincreasedPlt0.05.ComparedwiththepositivecontrolgrouptheexpressionofCaspase-3andBaxsignificantlydecreasedinJHJMandJLgroupsPlt0.05andtheexpressionofBcl-2significantlyincreasedinJHandJMgr