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].现代生物医学进展英文版,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 |
Received:September 20, 2018 Revised:October 12, 2018 |
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) |
Author Name | Affiliation | E-mail | MA Hui-min | Department of Endocrinology, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210011, China | 291645580@qq.com | WEI Xiao | Endocrine and Diabetes Center, Jiangsu Province Hospital on Integration of Chinese and Western Medicine, Nanjing, Jiangsu, 210028, China | | WEN Hong-hua | Department of Endocrinology, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210011, China | | WANG Zheng | Department of Endocrinology, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210011, China | | TANG Shan | Department of Endocrinology, Huai'an First People's Hospital, Huai'an, Jiangsu, 223001, China | | LIU Chao | Department of Endocrinology, Jiangsu Province Hospital on Integration of Chinese and Western Medicine, Nanjing, Jiangsu, 210028, China | | ZHU Qun | Department of Endocrinology, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210011, China | |
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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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