Sakai Hiroyasu, Sagara Atsunobu, Matsumoto Kenjiro, Jo Ara, Hirosaki Akiko, Takase Kazuhide, Sugiyama Ryoto, Sato Ken, Ikegami Daigo, Horie Syunji, Matoba Motohiro, Narita Minoru
Department of Pharmacology, Hoshi University, Tokyo, Japan; Division of Pharmacy Professional Development and Research, Hoshi University, Tokyo, Japan.
Department of Pharmacology, Hoshi University, Tokyo, Japan.
Pharmacol Res. 2014 Sep;87:71-9. doi: 10.1016/j.phrs.2014.05.012. Epub 2014 Jun 25.
Diarrhea is a common side effect experienced by cancer patients undergoing clinical chemotherapy, such as with 5-fluorouracil (5-FU). However, the precise mechanisms underlying 5-FU-induced diarrhea remain unclear. In the present study, we examined the role of neutrophil in 5-FU-induced diarrhea. Mice were given 5-FU (50mg/kg, i.p.) daily for 4 days. Sivelestat sodium (100 or 300 mg/kg, i.p., neutorophil elastase inhibitor) or SB225002 (3 or 9 mg/kg, i.p., CXCR2 antagonist) was administered before the administration of 5-FU. Gene expression levels of aquaporin (AQP) 4 and 8, CXCL1, CXCL2, CXCL3, neutrophil elastase (Elane) and myeloperoxidase (MPO) in the colon were examined by real-time RT-PCR. The neutrophil (Ly-6G positive cell) number in the mucosa of colon was measured by flow-cytometric analysis. Administration of 5-FU induced diarrhea and decreased the expression levels of AQP 4 and 8 in the colon. Under the present conditions, the expression levels of CXCL1, CXCL2, CXCL3, the neutrophil markers Elane and MPO, as well as Ly-6G-positive neutrophils, in the colon were significantly increased by 5-FU. Neutrophil recruitment with decreased levels of AQP 4 and 8 were dramatically inhibited by either sivelestat sodium or SB225002. Furthermore, these reagents reduced the 5-FU-induced body weight loss and diarrhea. These findings provide evidence that neutrophil recruitment and neutrophil elastase may decrease the levels of AQP 4 and 8 in the colon of mice treated with 5-FU and contribute to the pathophysiology of 5-FU-induced body weight loss and diarrhea.
腹泻是接受临床化疗的癌症患者常见的副作用，如使用5-氟尿嘧啶（5-FU）时。然而，5-FU诱导腹泻的精确机制仍不清楚。在本研究中，我们研究了中性粒细胞在5-FU诱导腹泻中的作用。小鼠每天腹腔注射5-FU（50mg/kg），共4天。在给予5-FU之前，腹腔注射西维来司他钠（100或300mg/kg，中性粒细胞弹性蛋白酶抑制剂）或SB225002（3或9mg/kg，CXCR2拮抗剂）。通过实时RT-PCR检测结肠中aquaporin（AQP）4和8、CXCL1、CXCL2、CXCL3、中性粒细胞弹性蛋白酶（Elane）和髓过氧化物酶（MPO）的基因表达水平。通过流式细胞术分析测量结肠黏膜中的中性粒细胞（Ly-6G阳性细胞）数量。给予5-FU会导致腹泻，并降低结肠中AQP 4和8的表达水平。在当前条件下，5-FU可显著增加结肠中CXCL1、CXCL2、CXCL3、中性粒细胞标志物Elane和MPO的表达水平，以及Ly-6G阳性中性粒细胞的数量。西维来司他钠或SB225002均可显著抑制中性粒细胞募集以及AQP 4和8水平的降低。此外，这些试剂减轻了5-FU诱导的体重减轻和腹泻。这些发现证明，中性粒细胞募集和中性粒细胞弹性蛋白酶可能会降低5-FU处理小鼠结肠中AQP 4和8的水平，并导致5-FU诱导的体重减轻和腹泻的病理生理过程。
