四君子汤多糖对IEC-6细胞迁移、钾通道蛋白及膜电位的影响

目的:观察四君子汤多糖对小肠上皮IEC-6细胞迁移及多胺调控信号通路钾通道蛋白和膜电位的影响,探讨四君子汤修复胃肠黏膜损伤的作用机制。方法:划痕法建立IEC-6细胞迁移模型,观察正常培养及α-二氧甲基乌氨酸(DFMO,多胺合成抑制剂)或4-氨基吡啶(4-AP,钾通道抑制剂)负荷下四君子汤多糖对IEC-6细胞迁移的影响;分别以荧光定量PCR法和Western blot法检测四君子汤多糖对IEC-6细胞钾通道蛋白kv1.1 mRNA和蛋白表达的影响;流式细胞仪检测四君子汤多糖对IEC-6细胞膜电位的影响。结果:与空白对照组比较,四君子汤多糖(40、80、160 mg/L)可促进IEC-6细胞迁移,提高IEC-6细胞kv1.1 mRNA和蛋白表达水平及细胞膜电位超极化水平(P<0.05或P<0.01);与DFMO模型组比较,四君子汤多糖(40、80、160 mg/L)可逆转DFMO所致细胞迁移数减少、kv1.1 mRNA和蛋白表达的降低以及细胞膜电位去极化(P<0.05或P<0.01);与4-AP模型组比较,四君子汤多糖(20、40、80 mg/L)可逆转4-AP所致细胞迁移的抑制、kv1.1 mRNA和蛋白表达的降低(P<0.05或P<0.01)。结论:四君子汤多糖可促进IEC-6细胞迁移,作用机制与影响多胺调控信号通路钾通道蛋白表达和细胞膜电位有关。

染料木黄酮对大鼠胰岛素分泌的调控作用

我国糖尿病的发病率近年来呈现快速增长的趋势。2型糖尿病以胰岛素分泌相对不足和胰岛素抵抗为主要特征，因此，促进胰岛素分泌是临床治疗2型糖尿病的重要手段。染料木黄酮（genistein）主要来源于豆类植物，大量证据表明染料木黄酮对糖尿病、慢性缺氧、骨质疏松等有一定的预防和治疗作用［1－3］。研究显示，染料木黄酮可改善2型糖尿病大鼠的胰岛素抵抗［1］，但其对胰岛素分泌的作用及机制尚不明晰。本研究采用放免法测定胰岛素分泌，膜片钳测定胰岛β细胞钾离子通道电流等方法，观察染料木黄酮对大鼠胰岛素分泌的影响，并探讨其作用机制。

线粒体:新的细胞内药物作用靶点

线粒体除了作为细胞内能量生成的关键细胞器 ,还与细胞凋亡、自由基生成、脂质代谢等重要生化过程密切相关。近年来随着对线粒体结构和功能的深入研究 ,发现线粒体是多类药物的细胞内作用靶点 ,与相关药物之间存在广泛而复杂的相互作用。研究线粒体与相关药物之间的相互作用对于明确药物作用机制、研发新药和避免药物毒副作用等方面具有重要意义。

Effects of Mahuang(Herba Ephedra Sinica) and Wuweizi(Fructus Schisandrae Chinensis) medicated serum on chemotactic migration of alveolar macrophages and inters regions macrophages in rats

OBJECTIVE: To observe the effects of serum containing Mahuang(Herba Ephedra Sinica) or Wuweizi(Fructus Schisandrae Chinensis) on the migration of alveolar macrophages(AM) and interstitial macrophages(IM) from normal rats, and to analyze and compare the mechanisms leading to cell migration differences.METHODS: Rats were randomly divided into three groups: Mahuang(Herba Ephedra Sinica), Wuweizi(Fructus Schisandrae Chinensis), and blank serum. After treatment with the herbs, serum was extracted from the rats. AM and IM were isolated from normal rats and cultured. The effects of Mahuang(Herba Ephedra Sinica) and Wuweizi(Fructus Schisandrae Chinensis) medicated serum on normal rat AM and IM chemotactic migration were determined by transwell assays. The CC chemokine receptor(CCR) 2, CCR5, voltage-gated Kvl. 3 K^+channel(Kv1. 3), and voltage-gated Kvl. 5 K^+channel(Kv1. 5) protein levels were analyzed by western blotting.RESULTS: The migration quantities of AM and IM in the Mahuang(Herba Ephedra Sinica) and Wuweizi(Fructus Schisandrae Chinensis) medicated serum groups were significantly higher than those in the blank serum group(P < 0.01). Compared with the Wuweizi(Fructus Schisandrae Chinensis) medicated serum group, the migration quantity of cultured rat AM in the Mahuang(Herba Ephedra Sinica) medicated serum group was significantly increased(P <0.01). Meanwhile, compared with the Mahuang(Herba Ephedra Sinica) medicated serum group, the migration quantity of cultured rat IM in the Wuweizi(Fructus Schisandrae Chinensis) medicated serum group was significantly increased(P < 0.01).CCR2, CCR5, Kv1. 3, and Kv1. 5 proteins were expressed on the AM cell surface, and showed significantly higher expression in the Mahuang(Herba Ephedra Sinica) medicated serum group compared with the Wuweizi(Fructus Schisandrae Chinensis)medicated serum group. In contrast, CCR5, Kv1.3,and Kv1.5 proteins were expressed on the IM cell surface, and showed significantly higher expression in the Wuweizi(Fructus Schisandrae Chinensis)medicated serum group compared with the Mahuang(Herba Ephedra Sinica) medicated serum group.CONCLUSION: Mahuang(Herba Ephedra Sinica)and Wuweizi(Fructus Schisandrae Chinensis) can promote AM and IM migration ability, with Mahuang(Herba Ephedra Sinica) targeting AM more apparently and Wuweizi(Fructus Schisandrae Chinensis) targeting IM more apparently. The mechanism may be that, by stimulating cells, Mahuang(Herba Ephedra Sinica) and Wuweizi(Fructus Schisandrae Chinensis) promote expression of CCR2 and CCR5 receptors on the AM and IM cell surface,which pass signals to Kvl.3 and Kvl.5 ion channels,leading to changes in the cytoskeleton, and ultimately promoting chemotactic cell migration.