炎症性肠病患者P2X7嘌呤受体及肥大细胞类胰蛋白酶表达研究
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摘要
目的探讨P2X7嘌呤受体及肥大细胞类胰蛋白酶在炎症性肠病(inflammatory bowel disease, IBD)患者肠组织及血清中的表达及临床意义。方法收集2016年12月至2018年3月于郑州大学第二附属医院住院的80例活动期IBD患者的肠组织活检石蜡包埋标本、血清样本及粪钙卫蛋白(FC)结果,其中溃疡性结肠炎(UC)患者58例,包括轻度活动性患者15例,中度活动性患者20例,重度活动性患者23例;克罗恩病(CD)患者22例,包括轻度活动性患者7例,中度活动性患者7例,重度活动性患者8例。另外收集40例结直肠癌患者行外科手术切除的正常断端肠组织作为对照,40名健康志愿者的血清样本作为对照。采用免疫组织化学法检测肠组织中P2X7嘌呤受体的表达,ELISA法检测血清类胰蛋白酶的含量。结果轻、中、重度UC组P2X7嘌呤受体的表达均高于对照组(χ~2=7.062,P=0.008;χ~2=18.983,P<0.05;χ~2=32.216,P<0.05),且重度UC组高于轻度UC组(χ~2=7.242,P=0.007)。轻、中、重度CD组P2X7嘌呤受体的表达高于对照组(χ~2=5.223,P=0.022;χ~2=5.223,P=0.022;χ~2=11.161,P=0.001),CD各亚组比较差异无统计学意义(P>0.05)。类胰蛋白酶在各组血清中的表达差异有统计学意义(P<0.001),IBD组高于对照组,且重度IBD组高于轻、中度IBD组。另外,P2X7嘌呤受体与类胰蛋白酶呈显著正相关(UC:r=0.872,P<0.05;CD:r=0.866,P<0.05)。P2X7嘌呤受体与FC呈正相关(r=0.543,P<0.05)。结论 P2X7嘌呤受体可能通过激活肥大细胞增加类胰蛋白酶的表达参与IBD的发生和发展,检测其变化对疾病严重程度的评估具有重要临床意义。
Objective To explore the expressions of P2 X7 purinoceptor and mast cell tryptase and to analyze their correlation in the intestinal tissues and serum of patients with inflammatory bowel disease(IBD). Methods A total of 80 active IBD patients in the Second Affiliated Hospital of Zhengzhou University from Dec. 2016 to Mar. 2018 were collected. There were 58 cases of ulcerative colitis(UC), among which 15 patients were mild UC, 20 patients were moderate UC, 23 patients were severe UC. There were 22 cases of Crohn's disease(CD), among which 7 patients were mild CD, 7 patients were moderate CD, 8 patients were severe CD. Their paraffin-embedded specimens, serum samples and results of fecal calprotectin(FC) were collected. The normal intestinal mucosa tissues from surgical specimens of 40 colorectal cancer patients were collected as tissue controls, blood samples from 40 healthy volunteers were collected as serum controls. The expression of P2 X7 purinoceptor in the intestinal mucosa was detected by immunohistochemistry, and the levels of tryptase in serum were measured by ELISA. Results Compared with control group, the expressions of P2 X7 purinoceptor in mild, moderate and severe UC group were significantly increased(χ~2=7.062, P=0.008; χ~2=18.983, P<0.05; χ~2=32.216, P<0.05). The severe UC group was higher than the mild UC group(χ~2=7.242, P=0.007). The expressions of P2 X7 purinoceptor in mild, moderate and severe CD group were significantly increased(χ~2=5.223, P=0.022; χ~2=5.223, P=0.022; χ~2=11.161, P=0.001). There was no statistical difference among the CD sub-groups(P>0.05). The expression of tryptase in IBD group was significantly increased(P<0.001). The expression of tryptase in severe IBD group was higher than that in the mild and moderate IBD group. In addition, the expression of P2 X7 purinoceptor was positively correlated with tryptase(UC: r=0.872, P<0.05; CD: r=0.866, P<0.05). The expression of P2 X7 purinoceptor was positively correlated with FC(r=0.543, P<0.05). ConclusionP2 X7 purinoceptor may participate in the development and progression of IBD by activating mast cell to increase the expression of tryptase.
Objective To explore the expressions of P2 X7 purinoceptor and mast cell tryptase and to analyze their correlation in the intestinal tissues and serum of patients with inflammatory bowel disease(IBD). Methods A total of 80 active IBD patients in the Second Affiliated Hospital of Zhengzhou University from Dec. 2016 to Mar. 2018 were collected. There were 58 cases of ulcerative colitis(UC), among which 15 patients were mild UC, 20 patients were moderate UC, 23 patients were severe UC. There were 22 cases of Crohn's disease(CD), among which 7 patients were mild CD, 7 patients were moderate CD, 8 patients were severe CD. Their paraffin-embedded specimens, serum samples and results of fecal calprotectin(FC) were collected. The normal intestinal mucosa tissues from surgical specimens of 40 colorectal cancer patients were collected as tissue controls, blood samples from 40 healthy volunteers were collected as serum controls. The expression of P2 X7 purinoceptor in the intestinal mucosa was detected by immunohistochemistry, and the levels of tryptase in serum were measured by ELISA. Results Compared with control group, the expressions of P2 X7 purinoceptor in mild, moderate and severe UC group were significantly increased(χ~2=7.062, P=0.008; χ~2=18.983, P<0.05; χ~2=32.216, P<0.05). The severe UC group was higher than the mild UC group(χ~2=7.242, P=0.007). The expressions of P2 X7 purinoceptor in mild, moderate and severe CD group were significantly increased(χ~2=5.223, P=0.022; χ~2=5.223, P=0.022; χ~2=11.161, P=0.001). There was no statistical difference among the CD sub-groups(P>0.05). The expression of tryptase in IBD group was significantly increased(P<0.001). The expression of tryptase in severe IBD group was higher than that in the mild and moderate IBD group. In addition, the expression of P2 X7 purinoceptor was positively correlated with tryptase(UC: r=0.872, P<0.05; CD: r=0.866, P<0.05). The expression of P2 X7 purinoceptor was positively correlated with FC(r=0.543, P<0.05). ConclusionP2 X7 purinoceptor may participate in the development and progression of IBD by activating mast cell to increase the expression of tryptase.
引文
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[14] BISCHOFF S C.Mast cells in gastrointestinal disorders [J].Eur J Pharmacol,2016,778:139-145.DOI:10.1016/j.ejphar.2016.02.018.
[2] NEVES A R,CASTELOBRANCO M T,FIGLIUOLO V R,et al.Overexpression of ATP-activated P2X7 receptors in the intestinal mucosa is implicated in the pathogenesis of Crohn’s disease [J].Inflamm Bowel Dis,2014,20(3):444-457.DOI:10.1097/01.MIB.0000441201.10454.06.
[3] YOSUKE K,TAKEAKI A,TOMONORI N,et al.Extracellular ATP mediates mast cell-dependent intestinal inflammation through P2X7 purinoceptors [J].Nat Commun,2012,3(3):1034-1045.DOI:10.1038/ncomms2023.
[4] 中华医学会消化病学分会炎症性肠病学组.炎症性肠病诊断与治疗的共识意见(2012年,广州)[J].中华消化杂志,2012,32(12):818-831.DOI:10.3760/cma.j.issn.0254-1432.2012.12.002.
[5] DE-ROSSI T,KRAUSS N V,GIERA B,et al.Mast cell tryptase in sera of patients with Crohn’s disease and mastocytosis [J].Eur J Gastroenterol Hepatol,2009,21(3):273-277.DOI:10.1097/meg.0b013e3283218392.
[6] KURASHIMA Y,KIYONO H.New era for mucosal mast cells:their roles in inflammation,allergic immune responses and adjuvant development [J].Exp Mol Med,2014,46(3):e83-e89.DOI:10.1038/emm.2014.7.
[7] LANIS J M,KAO D J,ALEXEEV E E,et al.Tissue metabolism and the inflammatory bowel diseases [J].J Mol Med (Berl),2017,95(9):905-913.DOI:10.1007/s00109-017-1544-2.
[8] DI V F,DAL B D,SARTI A C,et al.The P2X7 Receptor in Infection and Inflammation [J].Immunity,2017,47(1):15-31.DOI:10.1016/j.immuni.2017.06.020.
[9] CHEN K,ZHANG J,ZHANG W,et al.ATP-P2X4 signaling mediates NLRP3 inflammasome activation:a novel pathway of diabetic nephropathy [J].Int J Biochem Cell Biol,2013,45(5):932-943.DOI:10.1016/j.biocel.2013.02.009.
[10] YOSHIDA K,ITO M,MATSUOKA I.Divergent regulatory roles of extracellular ATP in the degranulation response of mouse bone marrow-derived mast cells [J].Int Immunopharmacol,2017,43:99-107.DOI:10.1016/j.intimp.2016.12.014.
[11] WAREHAM K J,SEWARD E P.P2X7 receptors induce degranulation in human mast cells [J].Purinergic Signal,2016,12(2):235-246.DOI:10.1007/s11302-016-9497-4.
[12] BURRI E,BEGLINGER C,Von F S,et al.Fecal calprotectin and the clinical activity index are both useful to monitor medical treatment in patients with ulcerative colitis [J].Dig Dis Sci,2015,60(2):485-491.DOI:10.1007/s10620-014-3383-0.
[13] TOSHIHIRO M,KUMI I,MASAMICHI I,et al.Ceramide-CD300f binding suppresses experimental colitis by inhibiting ATP-mediated mast cell activation [J].Gut,2016,65(5):777-787.DOI:10.1136/gutjnl-2014-308900.
[14] BISCHOFF S C.Mast cells in gastrointestinal disorders [J].Eur J Pharmacol,2016,778:139-145.DOI:10.1016/j.ejphar.2016.02.018.