胰岛素样生长因子1对重症急性胰腺炎大鼠肺损伤细胞

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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【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likegrolydividedintothreegroups

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

3、bservation.CellapoptosisinlungtissueinedbyTUNELmethod,theexpressionsofbaxmRNAandbcl2mRNAamylase,histologicscoringoflunginjury,apoptoticindex,theexpressionsofbaxmRNAandbcl2mRNAmarkedlyincreased.Apoptoticcellsincreasedobviouslyundertransmissionelectronmicroscope.Asparedephase,serumamylase,histologic

4、scoringoflunginjury,apoptoticindex,expressionsofbaxmRNAinIGF1groupdecreasedsignificantl,RNAincreasedsignificantly.Apoptoticcellsdecreasedobviouslyintransmissionelectronmicroscopicobservation.CONCLUSION:Theapoptosisandexpressionofbaxandbcl2inlungtissuemightbeinvolvedinpathogenesisofthelunginjuryind

5、ucedbySAP.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：CAGGATCGAGCAGAGAGGA

6、TG；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及术后3h按

7、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.肺损伤病理评分各成员的表达水平和磷酸化水平来发挥其抗凋亡作用.在葡萄糖和氧化剂诱导的人和鼠肾小球膜细胞凋亡中，IGF1可通过升高bcl2/bax的比