Section of Digestive Diseases and Nutrition, Department of Medicine, University of Illinois at Chicago, Chicago, IL 60612, USA.
Am J Physiol Gastrointest Liver Physiol. 2013 Mar 15;304(6):G615-22. doi: 10.1152/ajpgi.00445.2012. Epub 2013 Jan 17.
Intestinal P-glycoprotein (Pgp/multidrug resistance 1), encoded by the ATP-binding cassette B1 gene, is primarily involved in the transepithelial efflux of toxic metabolites and xenobiotics from the mucosa into the gut lumen. Reduced Pgp function and expression has been shown to be associated with intestinal inflammatory disorders. Keratinocyte growth factor-2 (KGF2) has emerged as a potential target for modulation of intestinal inflammation and maintenance of gut mucosal integrity. Whether KGF2 directly regulates Pgp in the human intestine is not known. Therefore, the present studies were undertaken to determine the modulation of Pgp by KGF2 using Caco-2 cells. Short-term treatment of Caco-2 cells with KGF2 (10 ng/ml, 1 h) increased Pgp activity (~2-fold, P < 0.05) as measured by verapamil-sensitive [(3)H]digoxin flux. This increase in Pgp function was associated with an increase in surface Pgp levels. The specific fibroblast growth factor receptor (FGFR) antagonist PD-161570 blocked the KGF2-mediated increase in Pgp activity. Inhibition of the mitogen-activated protein kinase (MAPK) pathway by PD-98059 attenuated the stimulatory effects of KGF2 on Pgp activity. Small-interfering RNA knockdown of Erk1/2 MAPK blocked the increase in surface Pgp levels by KGF2. Long-term treatment with KGF2 (10 ng/ml, 24 h) also significantly increased PgP activity, mRNA, protein expression, and promoter activity. The long-term effects of KGF2 on Pgp promoter activity were also blocked by the FGFR antagonist and mediated by the Erk1/2 MAPK pathway. In conclusion, our findings define the posttranslational and transcriptional mechanisms underlying stimulation of Pgp function and expression by KGF2 that may contribute to the beneficial effects of KGF2 in intestinal inflammatory disorders.
肠道 P-糖蛋白(Pgp/多药耐药 1),由 ATP 结合盒 B1 基因编码,主要参与将有毒代谢物和外源性物质从黏膜转运到肠腔的跨上皮外排。已经证明,Pgp 功能和表达的降低与肠道炎症性疾病有关。角质细胞生长因子-2(KGF2)已成为调节肠道炎症和维持肠道黏膜完整性的潜在靶点。目前尚不清楚 KGF2 是否直接调节人类肠道中的 Pgp。因此,本研究旨在使用 Caco-2 细胞确定 KGF2 对 Pgp 的调节作用。用 KGF2(10ng/ml,1h)短期处理 Caco-2 细胞可增加 Pgp 活性(~2 倍,P<0.05),通过维拉帕米敏感的[3H]地高辛通量测量。Pgp 功能的这种增加与表面 Pgp 水平的增加有关。特定的成纤维细胞生长因子受体(FGFR)拮抗剂 PD-161570 阻断了 KGF2 介导的 Pgp 活性增加。用 PD-98059 抑制丝裂原活化蛋白激酶(MAPK)通路减弱了 KGF2 对 Pgp 活性的刺激作用。用 Erk1/2 MAPK 的小干扰 RNA 敲低阻断了 KGF2 引起的表面 Pgp 水平增加。用 KGF2(10ng/ml,24h)长期处理也显著增加了 PgP 活性、mRNA、蛋白表达和启动子活性。FGFR 拮抗剂和 Erk1/2 MAPK 通路也阻断了 KGF2 对 Pgp 启动子活性的长期影响。总之,我们的研究结果定义了 KGF2 刺激 Pgp 功能和表达的翻译后和转录机制,这可能有助于 KGF2 在肠道炎症性疾病中的有益作用。