AIM To clarify the roles of TWEAK and its receptor Fn14 in 5-fluorouracil(5-FU)-induced diarrhea.METHODS Diarrhea was induced in wild-type(WT), Fn14 knockout(KO), and IL-13 receptor(IL-13R)α1 KO BALB/c mice using a s...AIM To clarify the roles of TWEAK and its receptor Fn14 in 5-fluorouracil(5-FU)-induced diarrhea.METHODS Diarrhea was induced in wild-type(WT), Fn14 knockout(KO), and IL-13 receptor(IL-13R)α1 KO BALB/c mice using a single injection of 5-FU. Histological analysis, cytokine analysis, and flow cytometry was performed on ileal tissues and cells. Murine colon carcinomabearing mice were co-treated with an anti-TWEAK antibody and 5-FU. Embryonic fibroblast response to cytokines was also analyzed.RESULTS5-FU induced high Fn14 expression in epithelial cells. The severity of 5-FU-induced diarrhea was lower in Fn14 KO mice compared with WT mice. Administration of anti-TWEAK antibody reduced 5-FU-induced diarrhea without affecting the antitumor effects of 5-FU in vivo. 5-FU-induced expression of IL-13, IL-17 A, TNF-α, and IFN-γ in the ileum was Fn14 dependent. The severity of 5-FU-induced diarrhea was lower in IL-13Rα1 KO mice, indicating major role for IL-13 signaling via IL-13Rα1 in pathogenesis. We found that IL-13Rα2, an IL-13 neutralizing/cell protective receptor, was strongly induced by IL-33 in vitro and in vivo. IL-13Rα2 was upregulated in the ileum of 5-FU-treated Fn14 KO mice. Thus, the deletion of Fn14 upregulated IL-13Rα2 expression, which reduced IL-13 expression and activity. CONCLUSION Disruption of the TWEAK/Fn14 pathway affects several interconnected pathways, including those associated with IL-13, IL-33, and IL-13Rα2, to attenuate 5-FUinduced intestinal side effects.展开更多
目的探讨溶血磷脂酸(lysophosphatidic acid,LPA)对人肺成纤维细胞(HFL-1)白细胞介素-13受体α2(IL-13Rα2)表达和Ⅰ型胶原(collagen typeⅠ,COLAⅠ)合成的影响。方法细胞培养,显微镜观察LPA对HFL-1细胞生长的影响,提取HFL-1细胞总RNA,...目的探讨溶血磷脂酸(lysophosphatidic acid,LPA)对人肺成纤维细胞(HFL-1)白细胞介素-13受体α2(IL-13Rα2)表达和Ⅰ型胶原(collagen typeⅠ,COLAⅠ)合成的影响。方法细胞培养,显微镜观察LPA对HFL-1细胞生长的影响,提取HFL-1细胞总RNA,用RT-PCR检测LPA对HFL-1细胞IL-13Rα2 mRNA和COLA I mRNA表达的影响。用免疫组化方法检测LPA和白细胞介素-13(interleukin-13,IL-13)对HFL-1细胞COLA I蛋白表达的影响。图像分析RT-PCR电泳条带和免疫组化图像,统计学处理。结果①LPA诱导HFL-1细胞IL-13Rα2 mRNA的表达,并呈时间依赖性。②1μmol.L-1 LPA刺激HFL-1细胞,IL-13Rα2 mR-NA的表达增强。③LPA作用HFL-1细胞,对IL-13Rα1的表达无影响。④LPA抑制HFL-1细胞COLA I mRNA的表达,IL-13促进COLA I mRNA的表达,LPA+IL-13混合组COLA I mRNA表达水平比LPA组高,比IL-13组低。⑤免疫组化结果与RT-PCR结果一致。结论溶血磷脂酸诱导人肺成纤维细胞表达IL-13 Rα2 mRNA,并下调人肺成纤维细胞合成COLAⅠ。展开更多
基金Supported by the Grants-in-Aid for Scientific Research,No.[B]5H04503 and No.[C]25460965,No.16K09299 from the Ministry of Education,Culture,Sports,Science,and Technologythe National Center for Global Health and Medicine,No.23-101,No.25-104,No.26-110,No.26-117,and No.27-1406the MEXT-Supported Program for the Strategic Research Foundation at Private Universities for Waseda University
文摘AIM To clarify the roles of TWEAK and its receptor Fn14 in 5-fluorouracil(5-FU)-induced diarrhea.METHODS Diarrhea was induced in wild-type(WT), Fn14 knockout(KO), and IL-13 receptor(IL-13R)α1 KO BALB/c mice using a single injection of 5-FU. Histological analysis, cytokine analysis, and flow cytometry was performed on ileal tissues and cells. Murine colon carcinomabearing mice were co-treated with an anti-TWEAK antibody and 5-FU. Embryonic fibroblast response to cytokines was also analyzed.RESULTS5-FU induced high Fn14 expression in epithelial cells. The severity of 5-FU-induced diarrhea was lower in Fn14 KO mice compared with WT mice. Administration of anti-TWEAK antibody reduced 5-FU-induced diarrhea without affecting the antitumor effects of 5-FU in vivo. 5-FU-induced expression of IL-13, IL-17 A, TNF-α, and IFN-γ in the ileum was Fn14 dependent. The severity of 5-FU-induced diarrhea was lower in IL-13Rα1 KO mice, indicating major role for IL-13 signaling via IL-13Rα1 in pathogenesis. We found that IL-13Rα2, an IL-13 neutralizing/cell protective receptor, was strongly induced by IL-33 in vitro and in vivo. IL-13Rα2 was upregulated in the ileum of 5-FU-treated Fn14 KO mice. Thus, the deletion of Fn14 upregulated IL-13Rα2 expression, which reduced IL-13 expression and activity. CONCLUSION Disruption of the TWEAK/Fn14 pathway affects several interconnected pathways, including those associated with IL-13, IL-33, and IL-13Rα2, to attenuate 5-FUinduced intestinal side effects.
文摘目的探讨溶血磷脂酸(lysophosphatidic acid,LPA)对人肺成纤维细胞(HFL-1)白细胞介素-13受体α2(IL-13Rα2)表达和Ⅰ型胶原(collagen typeⅠ,COLAⅠ)合成的影响。方法细胞培养,显微镜观察LPA对HFL-1细胞生长的影响,提取HFL-1细胞总RNA,用RT-PCR检测LPA对HFL-1细胞IL-13Rα2 mRNA和COLA I mRNA表达的影响。用免疫组化方法检测LPA和白细胞介素-13(interleukin-13,IL-13)对HFL-1细胞COLA I蛋白表达的影响。图像分析RT-PCR电泳条带和免疫组化图像,统计学处理。结果①LPA诱导HFL-1细胞IL-13Rα2 mRNA的表达,并呈时间依赖性。②1μmol.L-1 LPA刺激HFL-1细胞,IL-13Rα2 mR-NA的表达增强。③LPA作用HFL-1细胞,对IL-13Rα1的表达无影响。④LPA抑制HFL-1细胞COLA I mRNA的表达,IL-13促进COLA I mRNA的表达,LPA+IL-13混合组COLA I mRNA表达水平比LPA组高,比IL-13组低。⑤免疫组化结果与RT-PCR结果一致。结论溶血磷脂酸诱导人肺成纤维细胞表达IL-13 Rα2 mRNA,并下调人肺成纤维细胞合成COLAⅠ。
