目的:Cenicriviroc(CVC)是一种CCR2/CCR5拮抗剂,已被证明可有效治疗炎症和纤维化疾病。我们的研究评估了其在结肠炎中的疗效。
方法:建立DSS诱导的急性和慢性结肠炎小鼠模型。通过疾病活动指数(DAI)评分评估CVC在结肠炎中的疗效,炎症和纤维化的组织学评估,和关键分子的表达测定。在体外实验中,将HT29细胞系暴露于TNFα以研究肠上皮细胞中的炎症信号传导。CCD-18Co结肠肌成纤维细胞和人原代结肠成纤维细胞被TGFβ1活化以模拟成纤维细胞活化。
结果:在HT29细胞中,CVC显著降低CCL5mRNA表达(P<0.01),但对CCL2无影响。此外,CVC降低下游CX3CL1(P<0.01)和TNFα表达(P<0.05),从而抑制炎症进展。在急性结肠炎小鼠中,如HE染色所示,CVC显着降低了DAI评分和血清TNFα水平(P<0.05),并减轻了结肠炎症。同时,CVC对肝脏没有不良影响,心,和老鼠的肾脏。另一方面,在慢性结肠炎的细胞模型中,CVC降低纤维化标志物的表达,包括FN,CTGF,α-SMA,和MMP9,并抑制TGFβ1诱导的纤维化活化(P<0.01)。此外,CVC减轻慢性结肠炎小鼠结肠纤维化。此外,CVC显著促进自噬,这有助于它对炎症的调节。
结论:CVC通过CCL5/CCR5信号显著抑制炎症,而不损害重要器官,并抑制慢性结肠炎的纤维化激活,提示其缓解结肠炎症和纤维化的巨大潜力。
OBJECTIVE: Cenicriviroc (CVC) is a CCR2/CCR5 antagonist that has been shown to be effective in the treatment of inflammatory and fibrotic diseases. Our study evaluated its efficacy in colitis.
METHODS: Mouse models of DSS-induced acute and chronic colitis were established. The efficacy of CVC in colitis was assessed by disease activity index (DAI) scores, histological assessment of inflammation and fibrosis, and expression assays of key molecules. In in vitro experiments, HT29 cell line was exposed to TNFα to study inflammatory signaling in intestinal epithelial cells. CCD-18Co colonic myofibroblasts and human primary colonic fibroblasts were activated by TGFβ1 to mimic fibroblast activation.
RESULTS: In HT29 cells, CVC significantly reduced mRNA expression of CCL5 (P < 0.01) but had no effect on CCL2. Furthermore, CVC reduced downstream CX3CL1 (P < 0.01) and TNFα (P < 0.05) expression, thereby inhibiting inflammatory progression. In acute colitis mice, CVC significantly reduced DAI scores and serum TNFα levels (P < 0.05) and attenuated colonic inflammation as shown by HE staining. Meanwhile, CVC had no adverse effects on the liver, heart, and kidney of mice. On the other hand, in cellular models of chronic colitis, CVC decreased the expression of fibrosis markers, including FN, CTGF, α-SMA, and MMP9, and inhibited TGFβ1-induced fibrotic activation (P < 0.01). In addition, CVC attenuated colonic fibrosis in chronic colitis mice. Moreover, CVC significantly promoted autophagy, which contributed to its regulation of inflammation.
CONCLUSIONS: CVC significantly inhibited inflammation through CCL5/CCR5 signaling without damaging vital organs and suppressed fibrotic activation in chronic colitis, suggesting its great potential to relieve colonic inflammation and fibrosis.