世界中医药精选2022年第11期隔物 [复制链接]

隔物灸调控缺氧诱导因子１α表达抑制慢性萎缩性胃炎大鼠糖酵解的机制研究

李琪１　吴梦蝶１　刘世敏２　萧有智３　王文佳１　马喆２　黄艳２　李灵杰１，２　李瞡１　刘慧荣１，２

（１上海中医药大学附医院，上海，２００４３７；２上海市针灸经络研究所，上海，２０００３０；３医院，上海，２０１１０７）

目的：观察慢性萎缩性胃炎（CAG）大鼠胃黏膜组织缺氧诱导因子-1α（HIF-1α）及糖酵解关键酶表达变化，探究隔物灸对CAG的作用及可能机制。方法：将雄性Wistar大鼠随机分为正常组和造模组，自由饮用mg／L浓度MNNG联合法进行CAG大鼠造模。鉴定模型成功后，造模组大鼠随机分为模型组、隔药饼灸组、隔姜灸组和西药组，每组６只。隔药饼灸组、隔姜灸组均取中脘、气海穴进行隔物灸干预，西药组给予叶酸悬浊液灌胃。采用HE染色法观察胃窦组织病理学变化，实时荧光定量PCR（RT-ｑPCR）法检测胃黏膜组织信号转导及转录激活因子3（STAT3）、丙酮酸激酶M2（PKM2）和HIF-1α的ｍRNA表达水平，免疫组织化学法检测胃黏膜组织STAT3、HIF-1α蛋白表达，比色法和ELISA法测定胃黏膜组织LDH、PKM2活性。结果：与正常组比较，模型组大鼠胃黏膜组织固有腺体结构紊乱，腺体减少，出现萎缩、肠化、假幽门腺化生和（或）异型增生。模型组大鼠胃黏膜组织HIF-1α、STAT3、PKM2ｍRNA表达上调（P＜0.05），STAT3、HIF-1α蛋白表达增加（P＜0.05），LDH、PKM2酶活性增加（P＜0.05）。与模型组比较，隔药饼灸组、隔姜灸组、西药组固有腺体排列较规整，固有腺体萎缩、肠上皮化生、异型增生程度减轻；隔药饼灸和隔姜灸均能下调大鼠胃黏膜组织中HIF-1α、STAT3ｍRNA与蛋白表达（P＜0.05），降低LDH、PKM2酶活性（P＜0.05）。结论：隔物灸中脘、气海穴可改善CAG大鼠胃窦组织形态学病变，修复胃黏膜损伤。抑制HIF-1α介导的异常糖酵解代谢水平可能是隔物灸法治疗CAG的效应机制之一。

艾灸疗法；隔药饼灸；隔姜灸；慢性萎缩性胃炎；缺氧诱导因子-1α；糖酵解

MechanismofIndirectMoxibustioninInhibitionofGlycolysisinRatswithChronicAtrophicGastritisbyRegulatingHIF-1αExpression

LIQi1,WUMengdie1,LIUShimin2,XIAOYouzhi3,WANGWenjia1,MAZhe2,HUANGYan2,LILingjie1,2,LIJing1,LIUHuirong1,2

(1YueyangHospitalofIntegratedTraditionalChineseandWesternMedicine,ShanghaiUniversityofTraditionalChineseMedicine,Shanghai,China;2ShanghaiResearchInstituteofAcupunctureandMeridian,Shanghai,China;3ShanghaiXinqidianRehabilitationHospital,Shanghai,China)

Abstract

ObjectiveoobservetheexpressionchangesofHIF-1αandglycolysisenzymesingastricmucosaofratswithchronicatrophicgastritis(CAG)andexplorethemechanismofindirectmoxibustiononCAG.Methods:MaleWistarratswererandomlydividedintoanormalgroupandanexperimentalgroup.TheCAGmodelwasinducedintheexperimentalgroupbyadministrationofMNNG(mg/L).Themodelratswerethenrandomlydividedintoamodelgroup,aherb-partitionedmoxibustion(HM)group,aginger-partitionedmoxibustion(GM)group,andawesternmedicine(WM)group,withsixratsineachgroup.“Zhongwan”(CV12)and“Qihai”(CV6)wereselectedforindirectmoxibustionintheHMgroupandtheGMgroup.ThefolicacidsuspensionwasadministeredtotheratsintheWMgroupbygavage.ThehistopathologicalchangesofgastricantrumwereobservedbyHEstaining.ThemRNAexpressionofSTAT3,PKM2andHIF-1αinthegastricmucosawasdetectedbyRT-qPCR,andtheproteinexpressionofSTAT3andHIF-1αwasdetectedbyimmunohistochemistry.TheactivitiesofLDHandPKM2weredeterminedbycolorimetryandELISA.Results:Comparedwiththenormalgroup,themodelgroupshowedthestructuraldisorderofinherentglands,reducedglands,atrophy,intestinalmetaplasia,pseudopyloricmetaplasia,and/ordysplasiainthegastricmucosa.Furthermore,themRNAexpressionofHIF-1α,STAT3,andPKM2inthegastricmucosawasup-regulated(P0.05),theproteinexpressionofSTAT3andHIF-1αwaselevated(P0.05),andtheenzymeactivitiesofLDHandPKM2werepotentiatedinthemodelgroup(P0.05).Comparedwiththemodelgroup,theHMgroup,theGMgroup,andtheWMgroupshowedtheregulararrangementofinherent

iththemodelgroup,theHMgroupandtheGMgroupdisplayeddown-regulatedmRNAandproteinexpressionlevelsofHIF-1αandSTAT3inthegastricmucosa(P0.05),andweakenedenzymeactivitiesofLDHandPKM2(P0.05).Conclusion:Indirectmoxibustionat“Zhongwan”(CV12)and“Qihai”(CV6)canimprovethehistologicalchangesofthegastricantrumandrepairgastricmucosadamageinratswithCAG,andthemechanismofindirectmoxibustioninthetreatmentofCAGmayberelatedtotheinhibitionofabnormalglycolysismediatedbyHIF-1α.

Keywords

Moxibustion;Herb-partitionedmoxibustion;Ginger-partitionedmoxibustion;Chronicatrophicgastritis;Hypoxia-induciblefactor-1α;Glycolysis