Glucose dominates the regulation of carboxylesterases induced by lipopolysaccharide or interleukin-6 in primary mouse hepatocytes
Glucose dominates the regulation of carboxylesterases induced by lipopolysaccharide or interleukin-6 in primary mouse hepatocytes
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葡萄糖主导原代小鼠肝细胞中脂多糖或白细胞介素 6 诱导的羧酸酯酶的调节
DOI：
10.1016/j.lfs.2014.07.019
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发表时间：
2014
期刊：
影响因子：
6.1
通讯作者：
Yang Jian
中科院分区：
文献类型：
--
作者：
Xiong Jing;Shang Wei;Wu Lili;Chen Ruini;Liu Wei;Ning Rui;Hu Gang;Yang Jian
药物配置紊乱与炎症和糖尿病有关，导致药物疗效和毒性的改变。羧酸酯酶是肝脏中主要的水解酶，催化许多治疗剂的水解生物转化。因此，我们研究葡萄糖如何影响白介素-6 （IL-6）和脂多糖（LPS）对羧酸酯酶的调节。主要方法培养小鼠原代肝细胞。采用Western blot或酶联免疫吸附法（ELISA）检测蛋白水平，用激光共聚焦扫描显微镜和流式细胞术检测孕激素X受体（PXR）的激活情况。羧酸酯酶活性用酶学和毒理学方法测定。与5.6 mM葡萄糖相比，升高的葡萄糖（11或25 mM）显著增加羧酸酯酶的表达。LPS和IL-6抑制了25 mM葡萄糖中羧酸酯酶的表达和活性，但刺激了5.6 mM葡萄糖中羧酸酯酶的表达和活性。羧酸酯酶表达的改变与核因子κB （NFκB）的激活不一致，但与妊娠X受体（PXR）的表达和激活重复。PXR活化的改变进一步通过亚细胞易位的差异及其靶基因多药耐药1 （MDR1）的表达得到证实。这表明在不同葡萄糖浓度下，PXR介导炎症介质对羧酸酯酶的调节，而不是炎症信号。意义本研究有助于阐明炎症介质对羧酸酯酶的调控作用，并提示糖尿病中羧酸酯酶参与的药物代谢和药物-药物相互作用应根据炎症反应和高血糖的强度重新评估。
AimsAltered drug disposition has been associated with inflammation and diabetes, leading to the alteration of drug efficacy and toxicity. Carboxylesterases are major hydrolytic enzymes in the liver, catalyzing the hydrolytic biotransformation of numerous therapeutic agents. Therefore, how glucose affects the regulation of carboxylesterases by interleukin-6 (IL-6) and lipopolysaccharide (LPS) were investigated.Main methodsPrimary mouse hepatocytes were cultured. Protein levels were measured by Western blot or enzyme linked immunosorbent assay (ELISA), while confocal laser scanning microscope and flow cytometry were used to confirm the activation of pregnane X receptor (PXR). Carboxylesterase activity was evaluated by enzymatic and toxicological assays.Key findingsElevated glucose (11 or 25 mM) significantly increased carboxylesterase expression compared to 5.6 mM glucose. Carboxylesterase expression and activity were inhibited by LPS or IL-6 in 25 mM glucose, but stimulated in 5.6 mM glucose. The altered expression of carboxylesterases was not consistent with the activation of nuclear factor kappa B (NFκB) but repeatedly with the expression and activation of pregnane X receptor (PXR). The altered activation of PXR was further evidenced by the differential subcellular translocation and the expression of its target gene multidrug resistance 1 (MDR1). It implies that PXR, instead of inflammatory signaling, mediates the regulation of carboxylesterases by inflammatory mediators in different glucose concentrations.SignificanceThe findings contribute to clarify the regulation of carboxylesterases by inflammatory mediators, and indicate that carboxylesterase-involved drug metabolism and drug–drug interactions in diabetes should be reevaluated according to the intensity of inflammatory reactions and hyperglycemia.