烟碱型乙酰胆碱受体激动剂GTS-21对鼠实验性结肠炎的影响

发布时间：2018-07-26 16:59

【摘要】：目的:通过建立葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导的鼠结肠炎模型,探讨alpha7烟碱型乙酰胆碱受体(alpha7 nicotinic acetylcholine receptor,α7n ACh R)激动剂GTS-21改善模型鼠结肠炎症的效果及可能机制。方法:8~10周龄SPF级雄性BALB/c小鼠随机分为正常对照(CON)组、DSS模型组、GTS-21治疗组(每组8只)。DSS模型组采用自由饮用含3.5%DSS水造模,治疗组自由饮用3.5%DSS水,同时给予GTS-21[10 mg/(kg·d),腹腔注射],共7 d,对照组予生理盐水,每天予各组鼠疾病活动性评分(DAI评分)。第8天处死小鼠,观察结肠黏膜组织大体改变并测其长度、湿重,HE染色后行肠组织学炎症(HI)评分;细胞因子芯片筛选变化的细胞因子;ELISA法进一步检测芯片筛选出的变化明显的细胞因子。结果:(1)DSS组鼠结肠长度变短[(8.22±0.37)cm vs.(11.65±0.30)cm,n=8,P0.001],DAI评分较正常组增高[(1.51±0.10)分vs.0分,n=8,P0.001],HI评分升高[(20.5±3.9)分vs.(0.9±0.4)分,n=8,P0.001],表明造模成功;(2)给予烟碱型乙酰胆碱受体激动剂GTS-21的DSS模型鼠,DAI评分下降[(0.25±0.10)分vs.(1.51±0.10)分,n=8,P0.001];结肠长度较DSS组改善[(9.42±0.32)cm vs.(8.22±0.37)cm,n=8,P0.05];HI评分减低[(7.5±2.0)分vs(20.5±3.9)分,n=8,P0.01],提示GTS-21能改善DSS诱导的鼠结肠炎症;(3)细胞因子芯片筛选实验结果表明,DSS组γ干扰素诱导单核细胞因子(monokine induced by IFN-γ,CXCL9/Mig)升高最明显,此外肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)、白细胞介素1β(interleukin 1β,IL-1β)、γ-干扰素(interferonγ,IFN-γ)也明显升高;(4)进一步ELISA检测芯片变化最明显的CXCL9/Mig,发现DSS组鼠血清CXCL9/Mig明显升高(P0.05),给予GTS-21后,血清CXCL9/Mig降低(P0.05)。结论:GTS-21能减轻实验结肠炎鼠肠道炎症,该作用可能与减低趋化因子CXCL9/Mig水平,进而减少炎症细胞的肠道聚集有关。
[Abstract]:Aim: to investigate the effect and mechanism of alpha7 nicotinic acetylcholine receptor (alpha7 nicotinic acetylcholine receptor, 伪 7n ACh R) agonist GTS-21) on the improvement of rat colitis induced by dextran sodium sulfate (dextran sulfate sodium). Methods male BALB/c mice of SPF grade 10 weeks old were randomly divided into two groups: normal control group (CON) group, n = 8). DSS model group (n = 8 in each group). The model group was made by free drinking of 3.5%DSS water, and the treatment group was free to drink 3.5%DSS water. At the same time, GTS-21 [10 mg/ (kg d), intraperitoneal injection] was given to the control group for 7 days. The control group was given normal saline, and the disease activity score (DAI score) was given to the rats in each group every day. On the 8th day, the mice were killed, the colonic mucosa was observed and the length of colonic mucosa was measured. The (HI) score of intestinal histologic inflammation was obtained after HE staining. Cytokines were screened by cytokine microarray and the cytokines were further detected by Elisa. Results: (1) the colonic length of DSS group was shorter [(8.22 卤0.37) cm vs. (11.65 卤0.30) cm ~ (-1) + 0.30) cm vs. (] Dai score was higher than that of normal group [(1.51 卤0.10) vs.0 score (n = 8) P _ (0.001)] hi score increased [(20.5 卤3.9) vs (0.9 卤0.4) vs. (0.9 卤0.4) min ~ (8) P _ (0.001)], and (2) the DSS model rats treated with nicotinic acetylcholine receptor agonist GTS-21 (0.25 卤0.10) scores decreased [(0.25 卤0.10) scores vs. (1. 51 卤0. 10) vs (1. 51 卤0. 10)] Compared with the DSS group, the colon length was improved [(9.42 卤0.32) cm vs. (8.22 卤0.37) cm P0.01] and the HI score was decreased [(7.5 卤2.0) points vs (20.5 卤3.9) min P0.01], indicating that GTS-21 could improve the DSS induced colitis in rats. (3) the results of cytokine microarray screening showed that the monocyte factor (monokine induced by IFN- 纬 CXCL9Mig increased most significantly in the DSS group. In addition, tumor necrosis factor 伪 (tumor necrosis factor alpha-TNF- 伪, interleukin-1 尾 (interleukin 1 尾 -IL-1 尾) and interferon 纬 (interferon 纬 -IFN- 纬) were also significantly increased. (4) the most obvious changes of CXCL9 / Mig were further detected by ELISA. It was found that the serum CXCL9/Mig in DSS group was significantly increased (P0.05), and the serum CXCL9/Mig was decreased after GTS-21 administration (P0.05). ConclusionTwo one GTS-21 can attenuate the intestinal inflammation in experimental colitis rats, which may be related to the reduction of chemokine CXCL9/Mig level and the reduction of intestinal aggregation of inflammatory cells.
【作者单位】： 南京医科大学第一附属医院消化科;
【基金】：国家自然科学基金(81270469)
【分类号】：R574.62
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