胆固醇致人血管内皮细胞某n某损伤的分子机制-(8204)

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1、遵义医学院硕士学位论文胆崮醇致人血管内皮细胞DNA损伤的分子机制TheMolecularMechanismofVascularEndothelialCellDNADamageInducedbyCholesterolAbstractobjective：Toexplorethemechanismofcholesterol—inducedHUVECsDNAdamagecausedbyincreasedROS：whethercholesterolimprovedROSincreasedinHUVECscausesDNAdamagemediatedbyMCPIPand

2、thisprocessismediatedbywhatmeans?Astoprovideatheoreticalbasisforin—depthunderstandingoftherelationshipofabnormallipidmetabolism，DNAdamageandatherosclerosis．Methods：1．CultivationandidentificationofHUVECs．2．ThedoseandtimeeffectsofcholesterolonexpressionofMCPIPmRNAandproteininHUVECswe

3、retestedbyreal—timePCRandwesternblot．3．MCPIPgeneinHUVECwassilentviathetransienttransfectionofsi-MCPlP．EvaluationofMCPIPsilenceefficiencyonmRNAandproteinlevelsbyusingreal—timePCRandwesternblot．4．7-H2AXfociformationwasdetectedbyimmunofluorescenceandv—H2AXprotein1evelbywesternblottode

4、terminetheconditionofDNAdamage．5．HUVECsDNAdamagewasmeasuredbysinglecellgelelectrophresisassay．6．ROSproductionwasassessedbyflowcytometry．7．TheexpressionofNox4andtotal—p47p“眩aswellasmembranep47p胁wasdeterminedbywesternblot．8．Immunohistochemicalstainingwasusedtoobservetheexpressionoft-

5、H2AXinAsmodelratthoracicaorta．Results：1．thecelllineswereSuccessfulculturedandimmunofluorescenceidentificationshowedthatthecell1inesisHUVECs．2．After24hoursofdifferentdosegroupsofcholester01incubation，real—timePCRresultsshowedthattheCholesterolconcentrationwhichWasgreaterthanorequalt

6、o6．25mg／LofeachdosegroupcomparedwithcontrolgroupdifferenceWasstatisticallysignificant(p

7、heincreaseofthetimetheexpressingquantitygraduallyincreased，comparedwiththecontrolgroupstatisticallysignificantdifferences(p<0．01)．ThedoseandtimeeffectsofcholesterolonexpressionofMCPIPproteininHUVECstestedbywesternblotshowedthattheresultswerebasicallyidenticalwiththetrendsofMCPIPmRN

8、Aexpression．3．MCPIPmRNAand