CAJ | 학술논문

目的探讨IgA肾病(IgAN)患者核心1β1,3半乳糖基转移酶伴侣蛋白(Cosmc)表达低下的原因,到底是基因突变还是外源性抑制.方法 40例IgAN患者、16例非IgAN肾小球肾炎患者、21例健康对照被纳入本研究.直接测序方法测定Cosmc基因外显子区域的核酸序列.免疫磁珠分离IgAN患者、非IgAN肾小球.肾炎患者和健康对照者外周血B淋巴细胞,并分别在RPMI-1640及RPMI-1640+脂多糖(LPS)中培养72 h.实时荧光RT-PCR方法测定Cosmc基因的mRNA表达水平.结果 (1)IgAN患者Cosmc基因外显子区域没有共同突变位点,仅在2例患者中发现2处错义突变和2处无意义突变,而且这些突变并不影响患者cosmc基因的表达.(2)IgAN患者外周血B淋巴细胞的基础Cosmc基因表达显著降低,为健康对照组的31%.(3)经RPMI-1640培养后,IgAN患者Cosmc基因的表达水平升高至基础值的219%.RPMI-1640+LPS组Cosmc基因的h调受到明显抑制.(4)健康对照Cosmc基因的表达不受RPMI-1640和LPS的影响.结论 IgAN患者外周血B淋巴细胞Cosmc基因表达低下可能与基因突变无关,而是受外源性抑制囚素影响所致.
목적 탐토IgA신병(IgAN)환자핵심1β1,3반유당기전이매반려단백(Cosmc)표체저하적원인,도저시기인돌변환시외원성억제.방법 40례IgAN환자、16례비IgAN신소구신염환자、21례건강대조피납입본연구.직접측서방법측정Cosmc기인외현자구역적핵산서렬.면역자주분리IgAN환자、비IgAN신소구.신염환자화건강대조자외주혈B림파세포,병분별재RPMI-1640급RPMI-1640+지다당(LPS)중배양72 h.실시형광RT-PCR방법측정Cosmc기인적mRNA표체수평.결과 (1)IgAN환자Cosmc기인외현자구역몰유공동돌변위점,부재2례환자중발현2처착의돌변화2처무의의돌변,이차저사돌변병불영향환자cosmc기인적표체.(2)IgAN환자외주혈B림파세포적기출Cosmc기인표체현저강저,위건강대조조적31%.(3)경RPMI-1640배양후,IgAN환자Cosmc기인적표체수평승고지기출치적219%.RPMI-1640+LPS조Cosmc기인적h조수도명현억제.(4)건강대조Cosmc기인적표체불수RPMI-1640화LPS적영향.결론 IgAN환자외주혈B림파세포Cosmc기인표체저하가능여기인돌변무관,이시수외원성억제수소영향소치.
Objective To clarify whether the Cosmc gene down-regnhtion in lgA nephropathy (IgAN) patients is resulted from genetic disorders or external suppressions. Methods Forty IgAN patients, 16 non-IgAN glomerulonephritis patients and 21 healthy controls were enrolled in the study. Genomie DNA was extracted and then Cosmc gene was amplified and sequenced. Peripheral B lymphoeytes were isolated and cultured with RPMI-1640 alone or plus lipopolysaecharide (LPS). The Cosmc mRNA expression levels at baseline, after RPMI-1640 culture or RPMI-1640+LPS treatment were measured respectively by real-time RT-PCR. Results (1) Only 2 missense mutations and 2 silent mutations were detected in coding frame region of Cosine gene in 2 IgAN patients. (2) The baseline Cosmc gene expression level was significantly lower in IgAN patients (31% of that in healthy controls) than that in healthy controls. (3) Relative quantification PCR indicated that Cosmc mRNA expression level was significantly increased (219% of baseline) after RPMI-1640 culture, and treatment of LPS could strongly inhibit this effect. (4) The Cosmc gene expression of healthy control was not affected by RPMI-1640 or LPS. Conclusion It is not genetic disorders but external suppression to cause the down-regulation of Cosmc gene mRNA expression in IgAN.