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培养基中的葡萄糖浓度会影响 TRPV1 和 CB1 受体的 mRNA 表达,并改变 PC12 细胞中辣椒素的毒性。

Glucose concentration in culture medium affects mRNA expression of TRPV1 and CB1 receptors and changes capsaicin toxicity in PC12 cells.

机构信息

Novel Drug Delivery Research Center, Faculty of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran.

Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.

出版信息

Iran J Basic Med Sci. 2014 Sep;17(9):673-378.

Abstract

OBJECTIVE S

Hyperglycemia is widely recognized as the underlying cause for some debilitating conditions in diabetic patients. The role of cannabinoid CB1 and vanilloid TRPV1 receptors and their endogenous agonists, endovanilloids, in diabetic neuropathy is shown in many studies. Here we have used PC12 cell line to investigate the possible influence of glucose concentration in culture medium on cytoprotective or toxic effects of a CB1 [WIN55 212-2 (WIN)], or TRPV1 [Capsaicin (CAS)] agonist.

MATERIALS AND METHODS

Cell viability was tested using the MTT assay. We have also measured TRPV1 and CB1 transcripts by real time reverse transcription-polymerase chain reaction while cells were grown in low (5.5 mM) and high (50 mM) glucose concentrations.

RESULTS

Real time PCR results indicated that high glucose medium increased (P<0.01) TRPV1 mRNA and decreased (P <0.001) that of CB1. Cell culture tests show that hyperglycemic cells are more vulnerable (Dose × Medium, F (3,63)=41.5, P<0.001) to the toxic effects of capsaicin compared to those grown in low glucose medium.

CONCLUSION

These findings propose that hyperglycemic conditions may result in neuronal cell death because of inducing a counterbalance between cytotoxic TRPV1 and cytoprotective CB1 receptors.

摘要

目的

高血糖被广泛认为是糖尿病患者一些虚弱状况的根本原因。大麻素 CB1 和香草素 TRPV1 受体及其内源性激动剂内源性大麻素,在糖尿病性神经病中的作用在许多研究中都有显示。在这里,我们使用 PC12 细胞系来研究培养基中葡萄糖浓度对 CB1 [WIN55212-2(WIN)]或 TRPV1 [辣椒素(CAS)]激动剂的细胞保护或毒性作用的可能影响。

材料和方法

使用 MTT 测定法测试细胞活力。当细胞在低(5.5mM)和高(50mM)葡萄糖浓度下生长时,我们还通过实时逆转录-聚合酶链反应测量 TRPV1 和 CB1 转录物。

结果

实时 PCR 结果表明,高葡萄糖培养基增加了 TRPV1 mRNA(P<0.01),并降低了 CB1 mRNA(P <0.001)。细胞培养试验表明,与在低糖培养基中生长的细胞相比,高血糖细胞对辣椒素的毒性作用更敏感(剂量×培养基,F(3,63)=41.5,P<0.001)。

结论

这些发现表明,高血糖条件可能导致神经元细胞死亡,因为它引起了细胞毒性 TRPV1 和细胞保护 CB1 受体之间的平衡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a302/4322151/485c2e69da7d/IJBMS-17-673_F1.jpg

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