逆转录－聚合酶链反应、狭缝印迹检测重症肌无力患者IL－2mRNA表达水平

Reversetranscription,polymerasechainreactiontechniqueandslotblotfordetectingexpressionofIL┐2mRNAinpatientswithmyastheniagravis

为了探讨重症肌无力的发病机制利用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）、狭缝印迹技术和非放射性地高辛配基（ＤＩＧ）ＤＮＡ标记和检测手段，采用分子生物学技术和免疫学反应相结合，定量检测了重症肌无力（ＭＧ）患者和健康对照外周血单个核细胞（ＰＢＭＣ）的白细胞介素－２信使核糖核酸（ＩＬ－２ｍＲＮＡ）表达水平。结果表明：ＲＴ－ＰＣＲ方法特异而灵敏，可检测出１００ｆｇ含量的ＤＮＡ。重症肌无力患者（ｎ＝３５）与健康对照（ｎ＝１５）的ＩＬ－２ｍＲＮＡ表达水平无明显差异（Ｐ＝０．１７）。用ＰＨＡ刺激ＰＢＭＣ，患者和对照的ＩＬ－２ｍＲＮＡ表达均明显增多。摘除胸腺或（和）肾上腺皮质激素治疗。

Thereversetranscription(RT),polymerasechainreaction(PCR)andslotblottech-niquewasusedforthequantificationofmessengerribonucleicacid(mRNA).Theexperimentsre-vealedthatthismethodwassensitiveinalowingthedetectionofthetargetcytokinemRNAdowntoatleast100fg.Wedeterminedtheinterleukine-2mRNAexpressionofperipheralbloodmononuclearcels(PBMC)from35patientswithmyastheniagravis(MG)and15healthycontrols.FolowingPHAtreatmentofPBMC,therewereincreasedIL-2mRNAexpressionsinbothgroups.Nodiffer-enceoftheIL-2mRNAlevelswasdetectedinthepatientsandthecontrolseitherwithPHAtreat-mentofPBMCorwithout.ButIL-2mRNAexpressionwasdecreasedinpatientsaftertherapywiththymectomyorglucocorticoidsascomparedtothatbeforethetherapyinthesamepatients.There-sultsshowedthattheRT-PCRandslotblottechniqueswerefoundtobesuitableforthedetectionofcytokineswhichareinthelowerexpressivelevels.Itwasconsideredthatimmunotherapyhadanin-hibitoryefectonIL-2mRNAexpressioninpatientswithMG.