高糖环境下Lipin1对神经元代谢组学的影响

doi:10.6040/j.issn.1671-7554.0.2022.1442

摘要/Abstract

摘要： 目的探讨高糖环境下磷脂酸磷酸酶(Lipin1)对PC12细胞代谢组学的影响。方法构建感染慢病毒的PC12细胞。用25 mmol/L正常糖浓度孵育感染空壳病毒的PC12细胞(WT组)和感染低表达Lipin1病毒的PC12细胞(D-WT组),用100 mmol/L高糖浓度孵育感染空壳病毒的PC12细胞(HG组)和感染过表达Lipin1病毒的PC12细胞(G-HG组),48 h后,使用蛋白免疫印迹法对Lipin1的表达进行验证(n≥6)。WT组、HG组和G-HG组处理48h,收集细胞。细胞进行液相色谱串联质谱,比较代谢组学差异(n=6)。结果与WT组相比,D-WT组和HG组的Lipin1表达明显降低(P<0.05);G-HG组与HG组相比,Lipin1表达明显升高(P<0.01)。以VIP>1,P<0.05,差异倍数(FC)>1.5或<0.65为差异代谢物筛选标准,与WT组相比,HG组谷氨酸明显下降;与HG组相比,G-HG组甘油二酯(DAG)、2-花生酰基甘油(2-AG)等明显上升。高糖孵育和调控Lipin1后的认知相关通路涉及到逆行内源性大麻素信号传导(ECS),谷氨酸能突触、长时程增强(LTP)和长时程抑制(LTD)等。结论高糖环境下谷氨酸降低可能是糖尿病脑病的发病机制,过表达Lipin1后可改善神经相关代谢物DAG、2-AG和相关通路ECS从而缓解认知障碍,Lipin1可能是糖尿病脑病关键治疗靶点之一。

关键词: Lipin1, 糖尿病脑病, 认知功能, 代谢组学

Abstract: Objective To investigate the effects of phosphatidic acid phosphatase(Lipin1)on the metabolomics of PC12 cells in high glucose environment. Methods PC12 cells infected with lentivirus were constructed. PC12 cells infected with empty capsid lentivirus(WT group)and underexpression Lipin1 lentivirus(D-WT group)were incubated with 25 mmol/L glucose. PC12 cells infected with empty capsid lentivirus(HG group)and overexpression Lipin1 lentivirus(G-HG group)were incubated with 100 mmol/L glucose. The expression of Lipin1 was determined with Western blotting after 48 h(n≥6). WT group, HG group and G-HG group were treated for 48h and the cells were collected and subjected to LC-MS/MS to compare the metabolomic differences(n=6). Results The Lipin1 expression was significantly higher in the WT group than in D-WT and HG groups(P<0.05), significantly higher in G-HG group than in HG group(P<0.01). When the screening criteria for differential metabolites were VIP>1, P<0.05, fold change(FC)>1.5 or <0.65, glutamic acid significantly decreased in HG group than in WT group; diacylglycerol(DAG)and 2-arachidacylglycerol(2-AG)significantly increased in G-HG group than in HG group. Cognitive-related pathway after high glucose incubation and regulation of Lipin1 involved the retrograde endocannabinoid signaling(ECS), glutamatergic synapse, long term enhancement(LTP)and long term inhibition(LTD). Conclusion Decreased glutamic acid in high glucose environment may explain the pathogenesis of diabetic encephalopathy. Overexpression of Lipin1 can improve neuro-related metabolites DAG and 2-AG and related pathway ECS to alleviate cognitive impairment. Lipin1 may be one of the key therapeutic targets for diabetic encephalopathy.

Key words: Lipin1, Diabetic encephalopathy, Cognitive function, Metabolomics

中图分类号:

R587.1

黄珊,娄能俊,韩晓琳,梁中昊,华梦羽,庄向华,陈诗鸿. 高糖环境下Lipin1对神经元代谢组学的影响[J]. 山东大学学报 (医学版), 2023, 61(2): 1-8.

HUANG Shan, LOU Nengjun, HAN Xiaolin, LIANG Zhonghao, HUA Mengyu, ZHUANG Xianghua, CHEN Shihong. Effects of Lipin1 on neuronal metabolomics in high glucose environment[J]. Journal of Shandong University (Health Sciences), 2023, 61(2): 1-8.

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