摘要
目的:检测人溃疡性结肠炎(Ulcerative Colitis,UC)患者结肠黏膜中CYP27B1和HBD-2变化,探讨维生素D在UC发病中的可能机制。方法:选择在河北医科大学第二医院消化内科诊断明确的活动期UC患者35例,应用免疫组织化学方法:、real-time PCR、western blot方法:测定正常对照、UC患者炎症和非炎症部位CYP27B1和HBD-2的表达和分布。结果:①HBD-2蛋白分布于UC炎症部位上皮细胞胞浆,炎性细胞核中也有表达。HBD-2总体表达在炎症部位较正常对照组显著升高(P<0.01);CYP27B1蛋白在正常组织、UC非炎症部位及炎症部位均表达。在上皮细胞主要位于近隐窝腺腔面细胞膜,在UC炎症部位淋巴细胞核中表达明显,炎症部位较正常对照组显著升高(P<0.05),UC炎症部位结肠上皮HBD-2和CYP27B1的表达呈正相关(r=0.710,P<0.01)。②HBD-2和CYP27B1的蛋白表达结果:显示:UC患者炎症部位二者的表达较正常黏膜均显著升高(P<0.01,P<0.05)。UC炎症部位HBD-2和CYP27B1的蛋白表达呈正相关(r=0.620,P<0.01)。③HBD-2和CYP27B1的mRNA表达结果:显示:UC患者炎症部位二者的表达较正常对照组均显著升高(P<0.01),且UC炎症部位结肠上皮HBD-2和CYP27B1的mRNA水平表达呈正相关(r=0.893,P<0.01)。结论:CYP27B1和HBD-2在病变部位显著高表达且呈正相关,CYP27B1可能参与了维生素D局部活化及调控防御素表达。
To investigate the expression of CYP27B1 and HBD-2 on the protein and transcription level in the tissue of colonic mucosa, and to explore the correlation between them, we collected a total of 35 patients with ulcerative colitis （UC）. The expressions of CYP27B1 and HBD-2 in colonic mucosa of UC patients and healthy people were detected by immunohistochemical staining. Then, the mRNA and protein expression level of CYP27B1 and HBD-2 were assessed by real-time fluorescent quantitative reverse transcription-polymerase chain reaction and western blot, respectively. The groups were arranged as follows： con group （normal colonic mucosa）; uninfl group （noninflammatory colonic mucosa of ulcerative colitis）; and infl group （inflammatory colonic mucosa of ulcerative colitis）. The results showed that HBD-2 protein expressed abundantly in the cytoplasm of epithelium with inflammation, and in the nucleus of inflammatory cells, respectively. The expression of HBD-2 in infl group was higher than that in con group （P〈0.01）. CYP27B1 was expressed in the three groups. The CYP27B 1 protein was located in the membrane of epithelium, near the crypt surface, and in the nucleus of inflammatory cells, respectively. And the CYP27B1 expression in infl group was significantly higher （P〈0.05）. The statistical analysis indicated that the expression of HBD-2 in inflammatory colonic mucosa was positively correlated with CYP27B1 （r = 0.710,P〈 0.01）. The data measured by western blot showed the protein expressions of HBD-2 and CYP27B1 in infl group were significantly higher than that in con group （P 〈0.01, P 〈0.05, respectively）. The statistical analysis indicated that the protein expression of HBD-2 in inflammatory colonic mucosa was positively correlated with CYP27B1 （r = 0.893,P〈 0.01）. The data measured by real time PCR showed the mRNA expressions of HBD2 and CYP27B1 in infl group were significantly higher than that in con group （P 〈0.01）. The statistical analysis indicated that the mRNA
出处
《免疫学杂志》
CAS
CSCD
北大核心
2010年第6期507-511,516,共6页
Immunological Journal
