CAJ | 학술논문

目的：探讨跨膜蛋白16A(TMEM16A)在顽固性功能性便秘患者结肠中的表达及意义。方法收集2014年2―6月间在南京军区总医院普通外科行外科手术切除的30例顽固性功能性便秘患者的结肠手术标本作为实验组(取材部位：乙状结肠全层标本)﹔选取30例经病理证实的非梗阻性结肠癌患者的手术标本作为对照组(取材部位：取距肿瘤至少5 cm以上的正常结肠组织)。分别采用免疫荧光染色、RT-PCR以及Western blot法检测TMEM16A和c-kit在结肠中的表达。结果实验组和对照组结肠标本中,均见TMEM16A蛋白及c-kit的表达,在结肠中的表达部位分布基本一致。实验组结肠组织中TMEM16A和c-kit的免疫荧光表达、蛋白含量及mRNA的表达均明显低于对照组(TMEM16A：吸光度值为1.84±0.25比3.65±0.32,灰度比为0.66±0.07比1.04±0.17,mRNA相对表达量为0.41±0.05比1.00﹔c-kit：吸光度值为3.38±0.24比5.06±0.31,灰度比为0.64±0.06比0.98±0.09,mRNA相对表达量为0.18±0.08比1.00﹔均P<0.05)。结论 TMEM16A在顽固性功能性便秘患者结肠组织中表达降低,可能通过调控c-kit的表达来调节结肠平滑肌的运动,并可能成为顽固性功能性便秘治疗的新靶点。
목적：탐토과막단백16A(TMEM16A)재완고성공능성편비환자결장중적표체급의의。방법수집2014년2―6월간재남경군구총의원보통외과행외과수술절제적30례완고성공능성편비환자적결장수술표본작위실험조(취재부위：을상결장전층표본)﹔선취30례경병리증실적비경조성결장암환자적수술표본작위대조조(취재부위：취거종류지소5 cm이상적정상결장조직)。분별채용면역형광염색、RT-PCR이급Western blot법검측TMEM16A화c-kit재결장중적표체。결과실험조화대조조결장표본중,균견TMEM16A단백급c-kit적표체,재결장중적표체부위분포기본일치。실험조결장조직중TMEM16A화c-kit적면역형광표체、단백함량급mRNA적표체균명현저우대조조(TMEM16A：흡광도치위1.84±0.25비3.65±0.32,회도비위0.66±0.07비1.04±0.17,mRNA상대표체량위0.41±0.05비1.00﹔c-kit：흡광도치위3.38±0.24비5.06±0.31,회도비위0.64±0.06비0.98±0.09,mRNA상대표체량위0.18±0.08비1.00﹔균P<0.05)。결론 TMEM16A재완고성공능성편비환자결장조직중표체강저,가능통과조공c-kit적표체래조절결장평활기적운동,병가능성위완고성공능성편비치료적신파점。
Objective To investigate the expression of TMEMl6A in the colon of intractable functional constipation patients and its clinical implications. Methods A total of 30 patients with intractable chronic functional constipation were selected as trial group (full thickness tissue of sigmoid colon), 30 patients with colon cancer as control group (distant tissues at least 5 cm away from cancer). Tissues from two groups were collected in our hospital from February 2012 to June 2014 and confirmed by pathological diagnosis. Immunofluorescence staining, RT-PCR and Western blot were performed to detect the mRNA and protein expression of TMEM16A and c-kit in colon. Results TMEM16A and c-kit protein expressions were observed in similar sites of colon tissues in two groups. Expressions of TMEM16A and C-kit in colon tissues detected by immunofluorescence, RT-PCR, and Western blotting were significantly lower in trial group than those in control group (TMEM16A: mean A 1.84±0.25 vs. 3.65±0.32, P<0.05, gray intensity ratio 0.66±0.07 vs. 1.04±0.17, P<0.05, relative mRNA 0.41±0.05 vs. 1.00, P<0.05﹔ c-kit: mean A 3.38±0.24 vs. 5.06±0.31, gray intensity ratio 0.64±0.06 vs. 0.98± 0.09, relative mRNA 0.18 ±0.08 vs. 1.00, all P<0.05). Conclusions TMEM16A expression in colon tissues of intractable functional constipation patients is significantly lower and may adjust the movement of colonic smooth muscle by regulating the c-kit expression. TMEMl6A may be used as a new candidate target for diagnosis and treatment of intractable functional constipation.