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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,P<0.001],DAI评分较正常组增高[(1.51±0.10)分vs.0分,n=8,P<0.001],HI评分升高[(20.5±3.9)分vs.(0.9±0.4)分,n=8,P<0.001],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,P<0.001];结肠长度较DSS组改善[(9.42±0.32)cm vs.(8.22±0.37)cm,n=8,P<0.05];HI评分减低[(7.5±2.0)分vs(20.5±3.9)分,n=8,P<0.01],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(P<0.05),给予GTS-21后,血清CXCL9/Mig降低(P<0.05)。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。
OBJECTIVE: To establish a murine model of colitis induced by dextran sulfate sodium (DSS) and to explore the effect of alpha7 nicotinic acetylcholine receptor (α7n ACh R) agonist GTS-21 Inflammatory effects and possible mechanisms. Methods: SPF male BALB / c mice aged 8 to 10 weeks were randomly divided into normal control (CON) group, DSS model group and GTS-21 treatment group (n = 8). The rats in DSS model group were given 3.5% DSS water for free drinking and 3.5% DSS water for drinking. GTS-21 [10 mg / (kg · d) Saline, disease activity score (DAI score) was given to each group on a daily basis. Mice were sacrificed on the 8th day, the colonic mucosa was observed and the length and wet weight were observed. The histological inflammatory score (HI) was determined by HE staining; the cytokines were screened by cytokine chip; further screening by ELISA The changes of obvious cytokines. Results: (1) The colon length of DSS group was shorter than that of the normal group [(8.22 ± 0.37) cm vs (11.65 ± 0.30) cm, n = 8, P <0.001] (20.5 ± 3.9) vs (0.9 ± 0.4), n = 8, P <0.001], indicating that the model was successful; (2) ) Decreased the DAI score [(0.25 ± 0.10) min vs. (1.51 ± 0.10) min, n = 8, P <0.001] in DSS model mice given nicotinic acetylcholine receptor agonist GTS-21; The improvement of the HI group was significantly better than that of the control group [(9.42 ± 0.32) cm vs. (8.22 ± 0.37) cm, n = 8, P <0.05] <0.01], suggesting that GTS-21 can improve DSS-induced colitis in mice; (3) Cytokine chip screening experiments showed that monocyte adhesion induced by IFN-γ (CXCL9 / Mig) In addition, the levels of tumor necrosis factor alpha (TNF-α), interleukin 1β (IL-1β) and interferon gamma (IFN-γ) (4) CXCL9 / Mig with the most obvious changes in the chip was detected by ELISA. The results showed that the CXCL9 / Mig of the DSS group was significantly increased (P <0.05). After the GTS-21 was administered, the CXCL9 / Mig was decreased (P <0.05). CONCLUSION: GTS-21 can reduce intestinal inflammation in experimental colitis rats, which may be related to the decrease of CXCL9 / Mig level and the reduction of inflammatory cell gut accumulation.