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

Document Type : Original Article

Authors

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

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

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.

Keywords


1. Kamenov ZA, Traykov LD. Diabetic somatic neuropathy. Adv Exp Med Biol 2012; 771:155-175.
2. Campos C. Chronic hyperglycemia and glucose toxicity: pathology and clinical sequelae. Postgrad Med  2012; 124:90-97.
3. Del Prato S. Glucose and LDL lowering: the need for intensive therapy. Curr Vasc Pharmacol 2012; 10:687-689.
4. Tomlinson DR, Gardiner NJ. Glucose neurotoxicity. Nat Rev Neurosci 2008; 9:36-45.
5. Zherebitskaya E, Akude E, Smith DR, Fernyhough P. Development of selective axonopathy in adult sensory neurons isolated from diabetic rats: role of glucose-induced oxidative stress. Diabetes 2009; 58:1356-1364.
6. Askwith T, Zeng W, Eggo MC, Stevens MJ. Oxidative stress and dysregulation of the taurine transporter in high-glucose-exposed human Schwann cells: implications for pathogenesis of diabetic neuropathy. Am J Physiol Endocrinol Metab 2009; 297:620-628.
7. Low PA, Nickander KK, Tritschler HJ. The roles of oxidative stress and antioxidant treatment in experimental diabetic neuropathy. Diabetes 1997; 46:38-42.
8. Vincent AM, Russell JW, Low P, Feldman EL. Oxidative stress in the pathogenesis of diabetic neuropathy. Endocr Rev 2004; 25:612-628.
9. Sugimoto K, Yasujima M, Yagihashi S. Role of advanced glycation end products in diabetic neuropathy. Curr Pharm Des 2008; 14:953-961.
10. Wada R, Yagihashi S. Role of advanced glycation end products and their receptors in development of diabetic neuropathy. Ann N Y Acad Sci 2005; 1043:598-604.
11. Oates PJ. Polyol pathway and diabetic peripheral neuropathy. Int Rev Neurobiol 2002; 50:325-392.
12. Mizuno K, Kato N, Makino M, Suzuki T, Shindo M. Continuous inhibition of excessive polyol pathway flux in peripheral nerves by aldose reductase inhibitor fidarestat leads to improvement of diabetic neuropathy. J Diabetes Complications 1999; 13:141-150.
13. Bae JS, Kim OK, Kim JM. Altered nerve excitability in subclinical/early diabetic neuropathy: evidence for early neurovascular process in diabetes mellitus? Diabetes Res Clin Pract 2011; 91:183-189.
14. Hafer-Macko CE, Ivey FM, Sorkin JD, Macko RF. Microvascular tissue plasminogen activator is reduced in diabetic neuropathy. Neurology 2007; 69:268-274.
15. Papanas N, Maltezos E. Cilostazol in diabetic neuropathy: premature farewell or new beginning? Angiology 2011; 62:605-608.
16. Obrosova IG. Diabetic painful and insensate neuropathy: pathogenesis and potential treatments. Neurotherapeutics 2009; 6:638-647.
17. Kostyuk E, Voitenko N, Kruglikov I, Shmigol A, Shishkin V, Efimov A, et al. Diabetes-induced changes in calcium homeostasis and the effects of calcium channel blockers in rat and mice nociceptive neurons. Diabetologia 2001; 44:1302-1309.
18. Huang TJ, Sayers NM, Fernyhough P, Verkhratsky A. Diabetes-induced alterations in calcium homeostasis in sensory neurones of streptozotocin-diabetic rats are restricted to lumbar ganglia and are prevented by neurotrophin-3. Diabetologia 2002; 45:560-570.
19. Shutov L, Kruglikov I, Gryshchenko O, Khomula E, Viatchenko-Karpinski V, Belan P, et al. The effect of nimodipine on calcium homeostasis and pain sensitivity in diabetic rats. Cell Mol Neurobiol 2006; 26:1541-1557.
20. Khomula EV, Viatchenko-Karpinski VY, Borisyuk AL, Duzhyy DE, Belan PV, Voitenko NV. Specific functioning of Cav3.2 T-type calcium and TRPV1 channels under different types of STZ-diabetic neuropathy. Biochim Biophys Acta 2013; 1832:636-649.
21. Gover TD, Moreira TH, Weinreich D. Role of calcium in regulating primary sensory neuronal excitability. Handb Exp Pharmacol 2009; 194:563-587.
22. Paschen W. Mechanisms of neuronal cell death: diverse roles of calcium in the various subcellular compartments. Cell Calcium 2003; 34:305-310.
23. Song J, Lee JH, Lee SH, Park KA, Lee WT, Lee JE. TRPV1 activation in primary cortical neurons induces calcium-dependent programmed cell death. Exp Neurobiol 2013; 22:51-57.
24. Ito N, Ruegg UT, Kudo A, Miyagoe-Suzuki Y, Takeda S. Capsaicin mimics mechanical load-induced intracellular signaling events: Involvement of TRPV1-mediated calcium signaling in induction of skeletal muscle hypertrophy. Channels  2013; 7:221-724.
25. Tsumura M, Sobhan U, Muramatsu T, Sato M, Ichikawa H, Sahara Y, et al. TRPV1-mediated calcium signal couples with cannabinoid receptors and sodium-calcium exchangers in rat odontoblasts. Cell Calcium 2012; 52:124-136.
26. Hu Y, Gu Q, Lin RL, Kryscio R, Lee LY. Calcium transient evoked by TRPV1 activators is enhanced by tumor necrosis factor-{alpha} in rat pulmonary sensory neurons. Am J Physiol Lung Cell Mol Physiol 2010; 299:483-492.
27. Samways DS, Khakh BS, Egan TM. Tunable calcium current through TRPV1 receptor channels. J Biol Chem 2008; 283:31274-31278.
28. Console-Bram L, Marcu J, Abood ME. Cannabinoid receptors: nomenclature and pharmacological principles. Prog Neuropsychopharmacol Biol Psychiatry 2012; 38:4-15.
29. McDowell TS, Wang ZY, Singh R, Bjorling D. CB1 Cannabinoid receptor agonist prevents NGF-induced sensitization of TRPV1 in sensory neurons. Neurosci Lett  2013; 551:34-38
30. Lisboa SF, Guimaraes FS. Differential role of CB1 and TRPV1 receptors on anandamide modulation of defensive responses induced by nitric oxide in the dorsolateral periaqueductal gray. Neuropharmacology 2012; 62:2455-2462.
31. Engel MA, Izydorczyk I, Mueller-Tribbensee SM, Becker C, Neurath MF, Reeh PW. Inhibitory CB1 and activating/desensitizing TRPV1-mediated cannabinoid actions on CGRP release in rodent skin. Neuropeptides  2011; 45:229-237.
32. Yang H, Wang Z, Capo-Aponte JE, Zhang F, Pan Z, Reinach PS. Epidermal growth factor receptor transactivation by the cannabinoid receptor (CB1) and transient receptor potential vanilloid 1 (TRPV1) induces differential responses in corneal epithelial cells. Exp Eye Res 2010; 91:462-471.
33. Kawamata T, Niiyama Y, Yamamoto J, Furuse S. Reduction of bone cancer pain by CB1 activation and TRPV1 inhibition. J Anesth 2010; 24:328-332.
34. Zhang F, Hong S, Stone V, Smith PJ. Expression of cannabinoid CB1 receptors in models of diabetic neuropathy. J Pharmacol Exp Ther 2007; 323:508-515.
35. Kamei J, Zushida K, Morita K, Sasaki M, Tanaka S. Role of vanilloid VR1 receptor in thermal allodynia and hyperalgesia in diabetic mice. Eur J Pharmacol 2001; 422 :83-86.
36. Someya A, Kunieda K, Akiyama N, Hirabayashi T, Horie S, Murayama T. Expression of vanilloid VR1 receptor in PC12 cells. Neurochem Int  2004; 4:1005-1010.
37. Mohammadi-Farani A, Sahebgharani M, Sepehrizadeh Z, Jaberi E, Ghazi-Khansari M. Diabetic thermal hyperalgesia: role of TRPV1 and CB1 receptors of periaqueductal gray. Brain Res 2010; 1328:49-56.
38. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001; 25:402-408.
 
39. Fowler CJ, Rojo ML, Rodriguez-Gaztelumendi A. Modulation of the endocannabinoid system: neuroprotection or neurotoxicity? Exp Neurol  2010; 224:37-47.
40. van der Stelt M, Di Marzo V. Cannabinoid receptors and their role in neuroprotection. Neuromolecular Med 2005; 7:37-50.
41. Veldhuis WB, van der Stelt M, Wadman MW, van Zadelhoff G, Maccarrone M, Fezza F, et al. Neuroprotection by the endogenous cannabinoid anandamide and arvanil against in vivo excitotoxicity in the rat: role of vanilloid receptors and lipoxygenases. J Neurosci 2003; 23:4127-4133.
42. Zhang F, Challapalli SC, Smith PJ. Cannabinoid CB(1) receptor activation stimulates neurite outgrowth and inhibits capsaicin-induced Ca(2+) influx in an in vitro model of diabetic neuropathy. Neuropharmacology  2009; 57:88-96.
43. Kruglikov I, Gryshchenko O, Shutov L, Kostyuk E, Kostyuk P, Voitenko N. Diabetes-induced abnormalities in ER calcium mobilization in primary and secondary nociceptive neurons. Pflugers Arch 2004; 448:395-401.
44. Ji RR, Samad TA, Jin SX, Schmoll R, Woolf CJ. p38 MAPK activation by NGF in primary sensory neurons after inflammation increases TRPV1 levels and maintains heat hyperalgesia. Neuron  2002; 36:57-68.
45. Amaya F, Oh-hashi K, Naruse Y, Iijima N, Ueda M, Shimosato G, et al. Local inflammation increases vanilloid receptor 1 expression within distinct subgroups of DRG neurons. Brain Res  2003; 963:190-196.
46. Vincent AM, Stevens MJ, Backus C, McLean LL, Feldman EL. Cell culture modeling to test therapies against hyperglycemia-mediated oxidative stress and injury. Antioxid Redox Signal 2005; 7:1494-1506.
47. Hong S, Wiley JW. Early painful diabetic neuropathy is associated with differential changes in the expression and function of vanilloid receptor 1. J Biol Chem 2005; 280:618-627.
48. Evans RM, Scott RH, Ross RA. Chronic exposure of sensory neurones to increased levels of nerve growth factor modulates CB1/TRPV1 receptor crosstalk. Br J Pharmacol  2007; 152:404-413.