CAJ | 학술논문

目的探讨SOX10、潘氏细胞发育及分泌防御素-5与先天性巨结肠相关性小肠结肠炎的关系.方法收集50例先天性巨结肠病变肠管,根据术前是否发生小肠结肠炎分为HAFC组(n=14)和HD组(n=36),并以20例正常结肠标本作对照组.采用免疫组织化学方法观察结肠中防御素-5蛋白质表达、潘氏细胞发育情况以及SOX10蛋白质表达情况.采用实时荧光定量PCR技术检测防御素-5 mRNA及Sox 10 mRNA表达情况.结果防御素-5在正常肠管中不表达,HAEC组和HD组在肠腺隐窝基底处呈不同程度阳性表达.但前者阳性区域平均光密度值明显增高(0.33±0.039比0.10±0.031,P＜0.05),HAEC组防御素-5mRNA亦呈显著增高趋势(2.72±0.80比0.78±0.21,P＜0.05).对结肠组织同层切片进行潘氏细胞特异性产物溶菌酶免疫组化染色发现,对照组肠管中除1例存在弱阳性外其他均无阳性表达.HD组和HAEC组结肠中同样在隐窝基底处存在溶菌酶阳性细胞,可鉴别为化生的潘氏细胞,但HAEC组在发生率(78.6%)和细胞个数(2.97±0.80)明显高于HD组(27.8%,0.43±0.85)(P＜0.05).SOX10免疫产物主要在结肠神经节细胞膜及胞浆中表达,对照组、HAEC组、HD组阳性区域的平均光密度值递减(0.75±0.041,0.61±0.048,0.35±0.025),差异具有统计学意义(P＜0.05),同时RT-PCR检测显示Sox10 mRNA在各组中的表达与蛋白质水平呈平行结果.结论 SOX10可能通过影响潘氏细胞发育及分泌防御素-5在先天性巨结肠相关性小肠结肠炎的发生发展中起一定作用.
목적 탐토SOX10、반씨세포발육급분비방어소-5여선천성거결장상관성소장결장염적관계.방법 수집50례선천성거결장병변장관,근거술전시부발생소장결장염분위HAFC조(n=14)화HD조(n=36),병이20례정상결장표본작대조조.채용면역조직화학방법관찰결장중방어소-5단백질표체、반씨세포발육정황이급SOX10단백질표체정황.채용실시형광정량PCR기술검측방어소-5 mRNA급Sox 10 mRNA표체정황.결과 방어소-5재정상장관중불표체,HAEC조화HD조재장선은와기저처정불동정도양성표체.단전자양성구역평균광밀도치명현증고(0.33±0.039비0.10±0.031,P＜0.05),HAEC조방어소-5mRNA역정현저증고추세(2.72±0.80비0.78±0.21,P＜0.05).대결장조직동층절편진행반씨세포특이성산물용균매면역조화염색발현,대조조장관중제1례존재약양성외기타균무양성표체.HD조화HAEC조결장중동양재은와기저처존재용균매양성세포,가감별위화생적반씨세포,단HAEC조재발생솔(78.6%)화세포개수(2.97±0.80)명현고우HD조(27.8%,0.43±0.85)(P＜0.05).SOX10면역산물주요재결장신경절세포막급포장중표체,대조조、HAEC조、HD조양성구역적평균광밀도치체감(0.75±0.041,0.61±0.048,0.35±0.025),차이구유통계학의의(P＜0.05),동시RT-PCR검측현시Sox10 mRNA재각조중적표체여단백질수평정평행결과.결론 SOX10가능통과영향반씨세포발육급분비방어소-5재선천성거결장상관성소장결장염적발생발전중기일정작용.
Objective To study the expression of SOX10 and Human α-defensins-5(HD-5 )in Hirschsprung's disease associated enterocolitis(HAEC) and expore the possible relationship between SOX10 and HAEC.Methods Fifty pathological colons of Hirschsprung's disease (HD) were divided into HAEC group (n = 14) and HD group(n = 36) according to the presence of preoperative enterocolitis,Twenty normal colons as control group.The protein and mRNA expression of HD-5 and SOX 10 were measured by immunohistochemical staining and real-time quantitative PCR Results Normal colons did not express HD-5 but positive expression of HD-5 was detected at the base of the crypts of Lieberkuhn in HAEC and HD groups in different degree.The mean optical density of HD-5 immunohistochemical staining (0.33 ± 0.039 vs 0.10 ± 0.031 )and HD-5 mRNA expression (2.72 ± 0.80 vs 0.78 ± 0.21 ) in HAEC group was apparently higher than those in HD group(P＜0.05).The expression of lysozyme,a specific product by Paneth Cell,on sequential sections was negative in control group except one sample.In HAEC group and HD group,positive expression of lysozyme can be seen in the crypts of Lieberkuhn,where the cells can be identified as Metaplastic Paneth cell,but the incidence (78.6% vs 27.8%) and the number of cells 2.97 ± 0.80 vs 0.43 ± 0.85) in HAEC group were obviously higher than those in HD group(2.97 ± 0.80 vs 0.43 ± 0.85) (P＜0.05).SOX10 was mainly located in the plasmalemma and cytoplasm of ganglion cell and its mean optical density in control group (0.75 ±0.041 ),HAEC group (0.61 ± 0.048)and HD group(0.35 ± 0.025) were decrement,the difference between three groups were statistically significant (P＜0.05).Meanwhile,Sox10mRNA detected by real-time quantitative PCR indicated the parallel result.Conclusions SOX10 may be an important factor in the pathogenesis and development of HAEC.