分泌型卷曲相关蛋白1在口腔黏膜下纤维化治疗前后唾液和龈沟液中的表达

Expression of secreted frizzled-related protein-1 in patient with oral submucous fibrosis

目的：检测口腔黏膜下纤维化（oral submucous fibrosis，OSF）患者局部注射曲安奈德和丹参前后唾液和龈沟液中分泌型卷曲相关蛋白1（secreted frizzled-related protein-1，SFRP1）质量浓度的变化及OSF患者颊黏膜组织中SFRP1的表达，初步探讨SFRP1在OSF发生发展中的作用。方法收集2013年4月至2015年12月中南大学湘雅医院口腔内科OSF中期男性患者20例为试验组，唾液和龈沟液收集来源于科室就诊的年龄匹配的口腔黏膜健康男性10例作为正常对照组。免疫组化组织标本正常口腔黏膜组织10例取自因口腔外伤或整形手术就诊于中南大学湘雅医院口腔颌面外科的患者。试验组利用简单随机法分为曲安奈德组和曲安奈德＋丹参联合治疗组连续治疗4周，每组各10例。利用酶联免疫吸附测定法检测受试者治疗前后唾液和龈沟液中SFRP1的质量浓度，并记录进食刺激痛视觉模拟评分分值和张口度。用免疫组织化学染色法检测OSF患者颊黏膜组织中SFRP1蛋白的表达。结果免疫组化结果显示SFRP1蛋白在口腔正常黏膜均呈阳性表达，阳性表达率10/10；OSF颊黏膜组织中SFRP1蛋白表达明显降低，阳性表达率为10%（2/20），与正常对照组相比差异有统计学意义（P〈0.01）。治疗前曲安奈德组唾液和龈沟液中SFRP1质量浓度分别为（105.8±27.6）和（84.7±33.2）ng/L，联合治疗组唾液和龈沟液中SFRP1质量浓度分别为（86.6±23.2）和（97.0±23.2）ng/L，均显著低于正常对照组[SFRP1质量浓度分别为（153.0±32.8）和（157.5±31.1）ng/L]（P〈0.01）。OSF患者颊黏膜组织中SFRP1蛋白表达与唾液和龈沟液中SFRP1质量浓度呈显著正相关（P〈0.01）。经连续4周治疗，曲安奈德组唾液和龈沟液中SFRP1质量浓度分别升高为（141.2±35.3）和（130.6±31.3）ng/L，联合治疗组SFRP1质量浓度分别升高为（148.5±65.9）和（123.04±27.4）ng/L ，均显著高于治疗前（P〈0.01）。结论单独应用曲安奈德或曲安奈德联合丹参局部治疗后临床疗效均改善，且唾液及龈沟液中SFRP1质量浓度显著升高。

Objective To investigate the concentrations and clinical significance of secreted frizzled-related protein-1（SFRP1） insaliva and gingival crevicular fluid of patients with oral submucous fibrosis （OSF） as well as the expression of SFRP1 in patients＆#39; OSF buccal mucosa. Methods Twenty OSF patients aged 20 to 40 years old were recruited and randomly divided into two experimental groups, of which were triamcinolone acetonide group and combined triamcinolone acetonide and salvia miltiorrhiza group, respectively. Ten healthy volunteers matchable in sex and age with the patients were recruited as control group. Concentrations of SFRP1 in saliva and gingival crevicular fluid were detected by enzyme-linked immunosorbent assay before and after a continuous treatment of 4 weeks. The visual analogue scale（VAS） pain scores and opening size were also recorded. The expression of SFRP1 in samples from OSF patients＆#39; buccal mucosa was also detected using immunohistochemical method. SPSS 16.0 was applied to analyze the results of the experiments. Results The concentrations of SFRP1 in saliva and gingival crevicular fluid before treatment were （105.8 ± 27.6） ng/L and （84.7 ± 33.2） ng/L in triamcinolone acetonide group, and （86.6 ± 23.2） ng/L and （97.0 ± 23.2） ng/L in combining group, which were both significantly lower（P〈0.01） than that in normal group（[153.0±32.8] ng/L and [157.5±31.1] ng/L）, respectively. The positive expression rate of SFRP1 in OSF group（10%[2/20]） was significantly lower than that of the control group（10/10）（P〈0.01）. After the treatment for 4 weeks, the concentrations of SFRP1 increased to （141.2 ± 35.3） ng/L and （130.6 ± 31.3） ng/L in triamcinolone acetonide group, and to （148.5 ± 65.9） ng/L and （123.0 ± 27.4） ng/L in combining group, which were both significantly higher than those of pre-treatment, respectively（P〈0.01）. Conclusions The concentrations of SFRP1 in saliva and gingival crevicular fluid of OSF patients, which positively corelated to the expression of SFRP1 in OSF patients＆#39; buccal mucosa, were significantly lower than that of normal individuals and increased significantly after treatments of local injections of triamcinolone acetonide only or combined with salvia miltiorrhiza.