| 马惠敏,韦 晓,温洪华,王 铮,唐 珊,刘 超,祝 群.低度增高的棕榈酸与脂多糖联合对胰岛β细胞活力的影响[J].,2019,19(9):1634-1637 |
| 低度增高的棕榈酸与脂多糖联合对胰岛β细胞活力的影响 |
| Effect of Low-grade Increase of Palmitate and Lipopolysaccharide on the Viability of Pancreatic β-cells |
| 投稿时间：2018-09-20 修订日期：2018-10-12 |
| DOI：10.13241/j.cnki.pmb.2019.09.007 |
| 中文关键词: 棕榈酸 脂多糖 中性神经酰胺酶 胰岛β细胞 |
| 英文关键词: Palmitate Lipopolysaccharide NCDase Pancreatic β-cells |
| 基金项目:江苏省自然科学基金项目(BK20151577） |
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| 中文摘要: |
| 摘要 目的：探讨低度增高的棕榈酸联合脂多糖对胰岛β细胞活力的影响及可能机制。方法：采用0.15 mmol/L棕榈酸和50 ng/mL脂多糖单独、联合刺激大鼠胰岛β细胞株INS-1细胞24h后，通过CCK8法检测细胞活力，Western blot方法检测细胞内神经鞘脂代谢的关键酶-中性神经酰胺酶(NCDase)的蛋白表达水平。进一步建立过表达NCDase基因重组质粒pEGFP-C3-NCDase并转染INS-1细胞后，用棕榈酸、脂多糖联合刺激24h，再通过CCK8法检测细胞活力。结果：与对照组相比，0.15 mmol/L棕榈酸或50 ng/mL脂多糖分别刺激INS-1细胞24h后对其细胞活力的影响无统计学意义(P>0.05)，但二者联合刺激可明显降低INS-1细胞的活力(P<0.05)。与对照组相比，单独棕榈酸或脂多糖刺激INS-1细胞后并不影响细胞内NCDase的蛋白表达，但联合刺激可显著下调NCDase的表达(P<0.05)；与pEGFP-C3+棕榈酸+脂多糖组相比，pEGFP-C3-NCDase明显削弱了棕榈酸联合脂多糖对INS-1细胞活力的抑制作用(P<0.05)。结论：低度增高的棕榈酸与脂多糖协同刺激可产生?茁细胞毒性作用，其机制可能与下调胰岛β细胞内NCDase表达有关。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the effect of low-grade increase of palmitate and lipopolysaccharide on the viability of pancreatic β-cells and its possible mechanism. Methods: The cell viability of rat pancreatic β-cell line INS-1 was detected by CCK8 assay after be- ing incubated with 0.15 mmol/L palmitate and 50 ng/mL lipopolysaccharide for 24 h respectively or together. The expression of NC- Dase protein in INS-1 cells was detected by Western blot assay. Stable clones of INS-1 cell line transfected with recombinant plasmids pEGFP-C3-NCDase and pEGFP-C3 vector were established. Then they were incubated with palmitate and lipopolysaccharide for 24 h. The cell viability was detected by CCK8 assay. Results: Compared with the control INS-1 cells, the viability of INS-1 cells was hardly af- fected by the stimulation of palmitate or lipopolysaccharide alone. However, the cell viability was significantly decreased after treatment with palmitate and lipopolysaccharide together(P<0.05). Compared with palmitate or lipopolysaccharide alone, the expression of NCDase protein in INS-1 cells was significantly decreased after incubated with palmitate and lipopolysaccharide(P<0.05). Compared with pEGFP-C3-palmitate plus lipopolysaccharide treatment group, NCDase overexpresion alleviated the cytotoxicity which was induced by palmitate and lipopolysaccharide in INS-1 cells (P<0.05). Conclusion: The synergistic effect of low-grade increase of palmitate and lipopolysaccharide on the cytotoxicity of β-cells is related to the down-regulation of NCDase protein in pancreatic β-cells. |
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