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1、Smad2反义寡核苷酸抑制高糖培养大鼠系膜细胞细胞外基质的分泌生壁銎查堡旦!鲞第2期ChinJNephrol,February2005,Vo121,No.2Smad2反义寡核苷酸抑制高糖培养大鼠系膜细胞细胞外基质的分泌郁胜强赖凌云马骥顾勇林善锬【摘要】目的观察Smad2反义寡核苷酸(ODN)对高糖培养的系膜细胞分泌纤连蛋白(FN),1V型胶原(Col1V)的影响,探讨Smad2在肾小球硬化中的作用,寻找延缓肾小球硬化的新方法.方法设计,合成Smad2起始密码子部位的正义,反义和随机ODN,用脂质体法将ODN瞬时转染大鼠系膜细胞,并进行高糖培养.观察Smad2mRNA及蛋
2、白的表达及其对系膜细胞细胞外基质(ECM)分泌的影响.结果转染Smad2反义ODN的大鼠系膜细胞,其Smad2mRNA及蛋白表达均降低,与高糖组,正义组和随机组相比差异有统计学意义(P<0.05),与正常对照组相比无统计学意义(P>0.05).高糖组,反义组,正义组和随机组培养上清液中FN含量(ng/m1)分别为84.19±6.8l,57.65+_5.05,78.72+_9.13,77.13±7.36;ColIV含量(ng/m1)分别为55.27+_4.75,22.3l±3.24,46.23+2.02,45.92±1.2l.反义ODN组FN,ColIV的含量
3、均减少,与高糖,正义,随机ODN组相比差异有统计学意义(P<0.05).结论反义Smad2ODN可抑制高糖刺激引起的系膜细胞Smad2FN和Col的表达上调,该研究结果为延缓肾脏纤维化提供了新的思路和治疗途径.【关键词】寡核苷酸类;细胞外基质;纤维化;Smad2TreatmentwithantisenseSmad20Iig0de0xynude0desreducesextracellularmatrixsecretioninratmeSanalcellsculturedwithhighglucoseYUSheng—qiang,LA,Ling—yun,MAJi,GUY
4、ong,L1NShah—yah.DivisionofNephrology,HuashanHospital,FudanUniversity,Shanghai200003,China【Abstract】ObjectiveToinvestigatetheeffectsofantisenseSmad2oligodeoxynudeotides(ODN)onfibronetin(FN)andcollagenIV(ColIV)secretionofratmesangialcellsculturedwithhighglucose,exploretheactionofSmad2inthe
5、glomerulosclerosisandtofindanewmethodtoretardtheprogressofglomerularfibrosis.Methods20一merantfisense,senseandrandomODNsweredesignedandsynthesizedthatwerephosphorothioatemodifiedtoincreasestability.TheantisenseODNencompassedtheATGoftheratSmad2gene.ODNwastranferredtransientlyintoratmesangi
6、alceilsthroughliposome.Ratceilsweretreatedwithhighglucose.mRNAandproteinofSmad2weredetectedbyRT—PCRandcytochemistry.FNandColIVwereexaminedbyELISA.ResultsAnfisenseODNsignificantlydecreasedmRNAandproteinexpressionofSmad2inratmesangialceilstreatedwithhighglucose(P<0.05).FNconcentrationsi
7、ntheculturemediumofhighslucosegroup,antisensegroup,sensegroupandrandomgroupwere(84.19+_6.81)ng/ml,(57.65±5.05)ng/ml,(78.72+_9.13)ng/mland(77.13+_7.36)ng/mlrespectively.ColIVconcentrationswere(55.27+_4.75)ng/na,(22.31±3.24)ng/na,(46.23+_2.02)ng/mland(45.92±1.21)ng/mlrespec
