胰岛素样生长因子1对重症急性胰腺炎大鼠肺损伤细胞凋亡的影响

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1、胰岛素样生长因子1对重症急性胰腺炎大鼠肺损伤细胞凋亡的影响【摘要】目的：探讨细胞凋亡和bax,bcl2基因表达在重症急性胰腺炎（SAP）肺损伤发病机制中的作用以及胰岛素样生长因子1(IGF1)对SAP肺损伤细胞凋亡的影响.方法：将:Toexploretherolesofcellapoptosisandexpressionofbaxandbcl2inlungtissueinthemechanismoflunginjuryinducedbysevereacutepancreatitis(SAP)andtheeffectsofinsulinlikegrolydividedintothreeg

2、roups:Controlgroup(n=24),SAPgroup(n=24)andIGF1group(n=24).Everygrouplydividedinto3timeunits(6,12,24h),eunit.ThemodelsofSAPtaurocholatesolutionintothebiliopancresticductofrats.Atthe6,12and24hafterestablishmentofmodels,serumamylase,histologicscoringoflunginjuryined.Thelungtissueissionelectronmicrosc

3、opicobservation.CellapoptosisinlungtissueinedbyTUNELmethod,theexpressionsofbaxmRNAandbcl2mRNAamylase,histologicscoringoflunginjury,apoptoticindex,theexpressionsofbaxmRNAandbcl2mRNAmarkedlyincreased.Apoptoticcellsincreasedobviouslyundertransmissionelectronmicroscope.Asparedephase,serumamylase,histo

4、logicscoringoflunginjury,apoptoticindex,expressionsofbaxmRNAinIGF1groupdecreasedsignificantl,RNAincreasedsignificantly.Apoptoticcellsdecreasedobviouslyintransmissionelectronmicroscopicobservation.CONCLUSION:Theapoptosisandexpressionofbaxandbcl2inlungtissuemightbeinvolvedinpathogenesisofthelunginju

5、ryinducedbySAP.IGF1canalleviatethelunginjurybyinhibitingapoptosisoflungtissuecells,inayplayanimportantrole.　　【Keya公司）；胰岛素样生长因子1（Biovision公司）；细胞凋亡原位检测试剂盒（Roche公司）；Trizol试剂、RTPCR相关试剂及引物（大连宝生物公司）.引物序列如下：bcl2（目的片段296bp）上游：CGGGCTGGGGATGACTTCTCT;下游：GCATCCCAGCCTCCGTTATCC；bax（目的片段452bp）上游：CAGGATCGAGCAGA

6、GAGGATG；下游：GTGAGGACTCCAGCCACAAAG；βactin（目的片段548bp）上游：ATGGATGACGTATCGCTG；下游：ATGAGGTAGTTGTCAGGT.　　1.2方法　　1.2.1动物分组将大鼠随机分为对照组、SAP组和IGF1组，每组24只.术前12h禁食、不禁水.用20g/L戊巴比妥钠按0.025mL/kg腹腔注射麻醉后常规备皮、消毒、铺巾，正中切口入腹.SAP组由胆胰管以0.1mL/min匀速逆行注入50g/L牛磺胆酸钠（0.01mL/kg）后逐层关腹；对照组自胆胰管内逆行注入等剂量生理盐水后逐层关腹；IGF1组造模同SAP组并于术前30min及

7、术后3h按50μg/kg分别皮下注射IGF1.3组在造模后分别于6,12,24h各取8只，右心室穿刺抽血，快速切取肺脏组织，测定各项指标.　　1.2.2血清淀粉酶测定用全自动生化分析检测仪测定.　　1.2.3肺组织学检查肺组织石蜡切片行HE染色，光镜观察.肺损伤病理评分参考文献［2］.　　1.2.4肺组织细胞凋亡测定采用TUNEL法测定，操作按试剂盒说明进行.同一切片高倍镜下取3个不同视野,分别