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目的:探讨缺氧诱导雪旺细胞膜水通道蛋白1(AQP1)表达机制及复活草提取物对缺氧损伤细胞的修复作用。方法:以不同浓度的CoCl_2(2、4、6、8、10μl/ml)培养细胞贴壁24h后,显微镜下观察细胞形态变化拍照并以Western Blot检测AQP1变化,确定最适浓度的CoCl_2进行后续试验。以不同浓度的复活草提取物(0.5、1.0、2.5、5.0、10.0μl/ml)培养贴壁细胞,24h后进行CCK-8检测。选取最适浓度的CoCl_2及复活草提取物进行后续试验,分为空白对照组、缺氧模型组、复活草提取物修复组观察7d,Western Blot检测7d的细胞AQP1及HIF-1α变化。结果:选取6μl/ml CoCl_2为最适浓度缺氧模型,细胞形态变化明显但对细胞生长未见明显抑制作用,AQP1表达含量最高。选取2.5μl/ml复活草提取物为最适浓度,能够明显促进细胞增殖。HIF-1α诱导雪旺细胞AQP1表达增加,复活草提取物能下调AQP1及HIF-1α的表达。结论:周围神经水肿与雪旺细胞AQP1的表达有明显相关性,细胞缺氧后HIF-1α诱导AQP1表达增加;复活草提取物能够明显下调缺氧的雪旺细胞中AQP1的变化从而减轻水肿。
AIM: To investigate the mechanism of hypoxia-induced Schwann cell membrane AQP1 expression and the effect of extract of Acacia extract on hypoxia-injured cells. Methods: The cells were cultured with different concentrations of CoCl_2 (2,4,6,8,10μl / ml) for 24 hours. Cell morphology was observed under a microscope and AQP1 was detected by Western Blot. The optimum concentration of CoCl_2 was determined and followed test. Adherent cells were cultured with different concentrations of resveratrol extract (0.5, 1.0, 2.5, 5.0 and 10.0μl / ml), and CCK-8 assay was performed after 24h. The optimal concentration of CoCl_2 and resveratrol extract were selected for follow-up test. The cells were divided into blank control group, hypoxia model group and resuscitation extract recovery group. The changes of AQP1 and HIF-1α were detected by Western Blot for 7 days. Results: The optimal concentration of CoCl 2 was 6μl / ml. The morphological changes of cells were obvious but no obvious inhibitory effect was found on the cell growth. The highest expression of AQP1 was found. Select 2.5μl / ml resveratrol extract as the optimal concentration, can significantly promote cell proliferation. HIF-1α induced an increase of AQP1 expression in Schwann cells, and the extract of Rejuvenated herbs could down-regulate the expression of AQP1 and HIF-1α. CONCLUSIONS: Peripheral neumatoid edema is significantly associated with the expression of AQP1 in Schwann cells. HIF-1α-induced AQP1 expression is increased after hypoxia. Rehabilitate extract can significantly reduce the changes of AQP1 in hypoxic Schwann cells to reduce edema.