补肾活血法对高糖条件下rgcs与视网膜müller细胞膜稳定性影响的实验研究-experimental study on the effect of bushen huoxue method on the stability of rgcs and retinal m ü ller cell membrane under high glucose condition.docxVIP

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补肾活血法对高糖条件下rgcs与视网膜müller细胞膜稳定性影响的实验研究-experimental study on the effect of bushen huoxue method on the stability of rgcs and retinal m ü ller cell membrane under high glucose condition.docx

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补肾活血法对高糖条件下rgcs与视网膜müller细胞膜稳定性影响的实验研究-experimental study on the effect of bushen huoxue method on the stability of rgcs and retinal m ü ller cell membrane under high glucose condition

ABSTRACTObjective:ToinvestigateprotectiveeffectofbushenhuoxuetherapyonRGCsandretinaMüllercellsculturedinvitrounderhighglucoseconditions，inaddition,themechanismofprotectiveactionwasalsopreliminarystudied.Methods:1.Theretinalcellsuspensionwasmadeofretinalfrom1dayoldSDrats，andthecellsuspensionwasscreenedbyseparatelyanti-ratsignal-regulatedprotein（CD172G）monoclonalantibodyandanti-ratThy-1.1monoclonalantibody，andfinallygotpurifiedRGCs；RGCswereinoculatedinretinalMüllercellsculturemediumsothattheco-culturemodelwasestablished，whichincludingbothRGCsandretinalMüllercells.2.ThentheRGCsandretinalMüllercellswereco-culturedinstablehighglucosecondition、fluctuationofglucoseconditionandnormalconditionseparately，allofthegroupswereintervenedbybloodserumincludingthemedicineofBushenHuoxueChinesemedicine.Theexperimentwasgroupedintonormalcontrolgroup,traditionalChinesemedicineinterveninggroup，stablehighglucosegroup，stablehighglucoseandtraditionalChinesemedicineinterveninggroup，glucosefluctuationgroup，glucosefluctuationandtraditionalChinesemedicineinterveninggroup.3.Theglutaminesynthetase(GS)activityandextracellularlactatedehydrogenase(LDH)leakageoftheco-culturecellsweremeasuredbyenzyme-linkedimmunosorbentassay(ELISA).AlldatawasanalyzedwithSPSS17.0.Results:At24hrsand48hrs,theLDHleakageofco-culturedcellsinthestablehighglucosegroup、glucosefluctuationgroupwassignificantlyincreased(Ρ0.05)comparedwiththenormalcontrolgroup.TheLDHleakageofco-culturedcellsinthestablehighglucosegroupwassignificantlyincreasedat72hrs(Ρ0.05)comparedwithits48hrs.At24hrs，theLDHleakageofco-culturedcellsinglucosefluctuationgroupwassignificantlyincreased(Ρ0.05)comparedwithstablehighglucosegroup.At24hrsand72hrs，theLDHleakageofco-culturedcellsinthestablehighglucoseandtraditionalChinesemedicineinterveninggroupwassignificantlyreduced(Ρ0.05)comparedwithstablehighglucosegroup.At24hrs，theLDHleakageofco-culturedcellsintheglucosefluctuationandtraditionalChinesemedicineinterveninggroupwassignificantlyreduced(Ρ0.05)comparedwiththeglu