糜酶抑制剂TY-51469在炎症性肠病治疗中的应用
Chymase inhibitor TY-51469 in therapy of inflammatory bowel disease.
作者信息
Liu Wei-Xin, Wang Ying, Sang Li-Xuan, Zhang Shen, Wang Ting, Zhou Feng, Gu Shou-Zhi
机构信息
Wei-Xin Liu, Ying Wang, Shen Zhang, Ting Wang, Feng Zhou, Department of Gastroenterology, First Affiliated Hospital, China Medical University, Shenyang 110001, Liaoning Province, China.
出版信息
World J Gastroenterol. 2016 Feb 7;22(5):1826-33. doi: 10.3748/wjg.v22.i5.1826.
AIM
To investigate the effect of chymase inhibitor TY-51469 in the therapy of inflammatory bowel disease and the underlying mechanism.
METHODS
Seventy-five healthy Sprague-Dawley rats were randomly assigned to one of the three groups (control group, model group and TY-51469 experiment group) and each group had twenty-five rats. The rats of the model group and experiment group were subjected to treatment with 3.5% dextran sulfate sodium (DSS) 10 mg/kg to induce colitis. The control group and model group were subjected to intraperitoneal injection of saline, while the experiment group was subjected to intraperitoneal injection of 10 mg/kg TY-51469 each day. Five rats of each group were sacrificed on 0, 7, 14, 21 and 28 d, respectively. The degree of inflammation was assessed by histopathological scoring; flow cytometry was performed to detect the proportion of CD4(+)CD25(+) Tregs in peripheral blood; colon tissues of rats were collected to measure mRNA and protein expression by PCR, Western blot and immunohistochemistry; serum levels of interleukin (IL)-10, transforming growth factor (TGF)-β1 and IL-17A were detected by ELISA.
RESULTS
The rats in the experiment group and model group had significantly more severe colitis than the ones in the control group (P < 0.05) before treatment on day 0; no significant difference was observed between the experiment group and model group (P > 0.05). After treatment with TY-51469, the rats in the experiment group had significantly less severe colitis compared with the model group on 7, 14, 21 and 28 d (P < 0.05). The proportion of CD4(+)CD25(+) Tregs was lower in the model group and experiment group than in the control group; the experiment group had a significantly higher proportion of CD4(+)CD25(+) Tregs than that in the model group (P < 0.05). The model group and experiment group demonstrated lower expression of Foxp3 than the control group; the experiment group had higher Foxp3 expression than the model group (P < 0.05). Cytokines IL-10, TGF-β1 and IL-17A were lower in the model group and experiment group than in the control group; the experiment group had higher expression than the model group (P < 0.05).
CONCLUSION
After treatment with chymase inhibitor TY-51469, the experiment group demonstrated more significantly reduced intestinal inflammation and higher expression of immune tolerance related cytokines (IL-10, TGF-β1, IL-17A) and Foxp3 which is specifically expressed in Tregs compared with the model group. Therefore, chymase inhibitor TY-51469 might ameliorate the progression of DSS-induced colitis possibly by increasing the expression of Tregs and cytokines.
目的
探讨糜酶抑制剂TY-51469对炎症性肠病的治疗作用及其潜在机制。
方法
将75只健康的Sprague-Dawley大鼠随机分为三组（对照组、模型组和TY-51469实验组），每组25只。模型组和实验组大鼠用10mg/kg 3.5%葡聚糖硫酸钠（DSS）处理以诱导结肠炎。对照组和模型组腹腔注射生理盐水，而实验组每天腹腔注射10mg/kg TY-51469。分别于第0、7、14、21和28天处死每组5只大鼠。通过组织病理学评分评估炎症程度；采用流式细胞术检测外周血中CD4(+)CD25(+)调节性T细胞（Tregs）的比例；收集大鼠结肠组织，通过PCR、蛋白质印迹法和免疫组织化学检测mRNA和蛋白表达；采用酶联免疫吸附测定（ELISA）法检测血清白细胞介素（IL）-10、转化生长因子（TGF）-β1和IL-17A水平。
结果
在第0天治疗前，实验组和模型组大鼠的结肠炎明显比对照组严重（P < 0.05）；实验组和模型组之间无显著差异（P > 0.05）。用TY-51469治疗后，在第7、14、21和28天，实验组大鼠的结肠炎比模型组明显减轻（P < 0.05）。模型组和实验组中CD4(+)CD25(+) Tregs的比例低于对照组；实验组中CD4(+)CD25(+) Tregs的比例明显高于模型组（P < 0.05）。模型组和实验组中叉头框蛋白3（Foxp3）的表达低于对照组；实验组中Foxp3的表达高于模型组（P < 0.05）。模型组和实验组中细胞因子IL-10、TGF-β1和IL-17A低于对照组；实验组中的表达高于模型组（P < 0.05）。
结论
与模型组相比，用糜酶抑制剂TY-51469治疗后，实验组肠道炎症明显减轻，免疫耐受相关细胞因子（IL-10、TGF-β1、IL-17A）和Tregs中特异性表达的Foxp3表达更高。因此，糜酶抑制剂TY-51469可能通过增加Tregs和细胞因子的表达来改善DSS诱导的结肠炎的进展。
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