慢性胃病不同中医证候患者胃黏膜基质金属蛋白酶-7及金属蛋白酶组织抑制剂-1的表达

Expressions of MMP-7 and TIMP-1 in Chronic Stomach Disease Patients with Different Syndromes of Chinese Medicine

目的观察慢性胃病不同中医证候胃黏膜基质金属蛋白酶-7(matrix metalloproteinase-7,MMP-7)、金属蛋白酶组织抑制剂-1(tissue inhibitor of metalloproteinase-1,TIMP-1)的表达,并探讨及其与Hp感染的关系。方法临床选取慢性胃病患者117例,并招募健康志愿者11名。根据中医证候将慢性胃病患者分为脾胃湿热证组(57例)、肝胃不和证组(30例)及脾气虚证组(30例),健康志愿者为正常对照组。采用美蓝染色和快速尿素酶法检测Hp感染情况;常规HE染色法观察炎症程度;免疫组织化学方法定性、定位检测MMP-7及TIMP-1蛋白水平表达。结果脾胃湿热证与肝胃不和证患者Hp感染率和感染程度相当,虽均呈略高于脾气虚证趋势,但差异无统计学意义(P>0.05)。与脾胃湿热组及脾气虚证组比较,肝胃不和证胃黏膜炎症程度更重(P<0.05),脾胃湿热证呈高于脾气虚证患者,但差异无统计学意义(P>0.05);胃黏膜炎症活动度呈脾胃湿热证>肝胃不和证>脾气虚证,差异有统计学意义(P<0.05)。与Hp阴性患者比较,脾胃湿热证、肝胃不和证及脾气虚证Hp阳性患者胃黏膜炎症活动度更重,脾胃湿热证及脾气虚证Hp阳性患者胃黏膜炎症程度更重,差异均有统计学意义(P<0.05)。与正常对照组比较,脾胃湿热证、肝胃不和证及脾气虚组患者胃黏膜TIMP-1表达水平升高(P<0.05,P<0.01),脾胃湿热证组Hp阴性患者MMP-7表达水平升高(P<0.05);与脾胃湿热证组Hp阴性患者比较,脾胃湿热证组Hp阳性患者MMP-7表达水平降低(P<0.05);与脾气虚证组比较,脾胃湿热证组TIMP-1蛋白表达水平降低(P<0.01)。胃黏膜炎症程度与MMP-7表达呈负相关,与TIMP-1表达呈正相关关系(P<0.01);Hp感染程度与胃黏膜MMP-7表达未见明显相关性(P>0.05),而与胃黏膜TIMP-1表达呈正相关(P<0.05);其中胃黏膜MMP-7表达与TIMP-1表达呈正相关(P<0.01)。结论胃黏膜炎症中MMP-7相对低水平表达及与TIMP-1平衡紊乱,可能是慢性胃病的发病机制之一;其在不同证候表达差异对"同病异证"微观研究有一定的阐发,且情绪波动可能也是慢性胃病发病的重要因素。

Objective To observe the expressions of matrix metalloproteinase-7 （MMP-7） and tissue inhibitor of metalloproteinase-1 （TIMP-1） in chronic stomach disease patients with different syn- dromes of Chinese medicine （CM）, and their relationships with Helicobacter pylori （ Hp ） infection. Meth- ods Totally 117 chronic stomach disease patients were recruited, and 11 healthy volunteers were also recruited. Chronic stomach disease patients were assigned to Pi-Wei dampness-heat syndrome （PWDHS, 57 cases）, disharmony of Gan and Wei syndrome （DGWS, 30 cases）, and Pi qi deficiency syndrome （PQDS, 30 cases） by syndrome typing. Healthy volunteers were recruited as the healthy con- trol group. Hp infection was detected using methylene blue dyeing and rapid urease test （RUT）. The de-gree of inflammation was observed by conventional HE staining. The protein expressions of MMP-7 and TIMP-1 were detected qualitatively and positioningly using immunohistochemical method. Results Pa- tients with PWDHS and patients with DGWS had equivalent Hp infection rate and degree. They showed a slightly increasing tendency than patients with PQDS, but with no statistical difference （P 〉0.05）. Com- pared with PWDHS and PQDS groups, more severe inflammation of mucosa occurred in patients with DG- WS （P 〈0.05）. More severe inflammation of mucosa occurred in patients with PWDHS than in those with PQDS, but with no statistical difference （P 〉0.05）. The severity of gastric mucosal inflammatory activity was sequenced from high to low as PWDHS, DGWS, PQDS, all with statistical difference （P 〈 0.05）. Compared with Hp negative patients, the gastric mucosal inflammatory activity was more severe in Hp positive patients with PWDHS, DGWS, PQDS. The gastric mucosal inflammatory activity was more se- vere in Hp positive patients with PWDHS and PQDS （P 〈0.05）. Compared with the healthy control group, the expression level of TIMP-1 in gastric mucosa increased in patients with PWDHS, DGWS, PQDS （P 〈0.05, P 〈0.01 ） ; the expression level of MMP-7 increased in Hp negative patients with PWDHS （P 〈 0.05）. Compared with lip negative patients with PWDHS, the expression level of MMP-7 decreased in Hp positive patients with PWDHS （P 〈0.05）. Compared with the PQDS group, the expression level of TIMP-1 decreased in the PWDHS group （P 〈0.01 ）. The severity of gastric mucosal inflammation was negatively correlated with the expression level of MMP-7, and positively correlated with the expression level of TIMP- 1 （P 〈0.01 ）.Hp infection degree was not obviously correlated with the expression level of MMP-7 in gas- tric mucosa （P 〉0.05）, but positively correlated with the expression level of TIMP-1 in gastric mucosa （P 〈0.05）. Of them, the expression level of MMP-7 in gastric mucosa was positively correlated with the expression level of TIMP-1 in gastric mucosa （P 〈0.01 ）. Conclusions Comparatively lower expression of MMP-7 in gastric mucosal inflammation and imbalanced expression of TIMP-1 might be two of the pathogeneses of chronic stomach disease. Their various expressions in different CM syndromes might have certain expositions for microscopic research on ＂different syndromes of the same disease＂. Emo- tional fluctuation miQht also be one of important factors for chronic stomach disease.