載脂蛋白A-Ⅰ突變體重組高密度脂蛋白對(duì)轉(zhuǎn)錄因子ATF3的影響.pdf

1、分類號(hào)：R543密級(jí)：不保密UDC：610學(xué)校代碼：11065碩士學(xué)位論文載脂蛋白載脂蛋白AI突變體重組高密度脂蛋白對(duì)轉(zhuǎn)錄因子突變體重組高密度脂蛋白對(duì)轉(zhuǎn)錄因子ATF3的影響影響王堰匯王堰匯指導(dǎo)教師侯琳教授學(xué)科專業(yè)名稱生物化學(xué)與分子生物學(xué)論文答辯日期2016.5.26AbstractObjective:ATF3asaTLRinducedrepressthatplaysanimptantroleininhibitionofspecifici

2、nflammatysignals.Highdensitylipoprotein(HDL)itsmajproteincomponentApoAIhassignificantantiinflammatyeffect.HereweinvestigateATF3effectofApolipoproteinAI(ApoAI)itscysteinemutantrecombinanthighdensitylipoproteinmethod:AI74A

3、IMAIWTAI74wasexpressedbythePETprokaryoticexpressionsystem.AfterpurificationbyaffinitychromatographytherecombinanthighdensitylipoproteinwaspreparedbyaddingDPPC.CμlturedRAW264.7cellsinflammatycellmodelinducedbyLPSstimμlati

4、on.Thenhighdensitylipoproteinintervention.DetectionoftheexpressionlevelofinflammatycytokinesIL6TNFαbyELISAkitobservationofthecellmphologicalchangesChangesintheexpressionlevelsofmRNAATF3proteinweredetectedinthecellsbyRTPC

5、Rwesternblotting.Results:CellmphologicalobservationfoundthatcomparedwiththenmalgrouptheLPSgrouptherHDL228groupcellmphologytendedtodevelopmentmacrophagedegradationrHDL74，rHDLMrHDLWThavedifferentprotectiveeffectsfLPSstimul

6、atedcells.ELISAdetectionshowedthatrHDL74rHDLWTrHDLMmakeTNαIL6expressionlevelsreturnedtoclosetotheblankcontrolgroupthelevelofTNFαIL6withrHDL228groupsincreasedsignificantly(P0.01).BlottingWesternresultsshowedthatcomparedwi

7、ththecontrolgroupLPSgrouprHDL228groupATF3expressionlevelsdecreasedthedifferencewassignificant(p0.01).TheexpressionlevelsofrHDL74rHDLMrHDLWTgroupweresignificantlyhigherthedifferencewassignificant(p0.05).Conclusion:Recombi

8、nanthighdensitylipoproteinrHDL74rHDLWTrHDLMpromotestheexpressionofATF3onmRNAproteinlevels.rHDL228downitsexpression.Postgraduatestudent:WangYanhui(BiochemistrymolecularBiology)DirectedbyProf:HouLinKeywds:rHDLsinflammatyli