Effects of docosahexenoic acid on large conductance Ca<sup>+</sup>-activated K<sup>+</sup> channels in retinal smooth muscle cells _Chinese Journal of Ocular Fundus Diseases

Authors：

Chen Xuan , Shao Jun , Xia Dayun , Wang Ruxing ,  Yao Yong

Wuxi People’s Hospital Affiliated to Nanjing Medical University, Wuxi 214023, China;

Corresponding author：

Yao Yong, Email: Pard1@126.com

Keywords：

Docosahexaenoic acids; Retinal artery; Myocytes, smooth muscle; Large-conductance calcium-activated potassium channels; Patch-clamp techniques

DOI：

10.3760/cma.j.issn.1005-1015.2017.03.017

Video：

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Objective To investigate the effects of docosahexenoic acid (DHA) on large conductance Ca²⁺-activated K⁺ (BK) channels in normal retinal artery smooth muscle cells (RASMCs). Methods Cultured human RASMCs (6 th-8 th generations) were used to patch clamp experiment. The open probabihties (NP₀) in BK channels with different concentrations (0.0, 1.0, 3.0, 5.0, 7.5, 10.0 μmol/L) of DHA were recorded by patch clamp technique in single channel configuration. RASMCs were intervened by different concentrations (0.0, 1.0, 5.0 μmol/L) of DHA as control group, low and high doses of DHA groups, respectively. The protein expressions of β subunit of BK channels in RASMCs from three groups were measured by Western blot. Results The NP₀ of BK channels were 0.044 4±0.001 2, 0.081 2±0.004 2, 0.209 0±0.006 1, 0.310 5±0.005 3, 0.465 0±0.007 8 and 0.497 7±0.014 5 with perfusate of 0.0, 1.0, 3.0, 5.0, 7.5, 10.0 μmol/L DHA. DHA activated BK channels in a dose-dependent manner (F=2.621,P＜0.05). There was no significant difference in the protein expression of control group, low and high doses of DHA groups (F=11.657,P＞0.05). Conclusion DHA can directly activate BK channels, no increasing in subunit expression of BK channels.

Citation： Chen Xuan, Shao Jun, Xia Dayun, Wang Ruxing, Yao Yong. Effects of docosahexenoic acid on large conductance Ca⁺-activated K⁺ channels in retinal smooth muscle cells . Chinese Journal of Ocular Fundus Diseases, 2017, 33(3): 295-297. doi: 10.3760/cma.j.issn.1005-1015.2017.03.017 Copy

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1. Galano JM, Lee JC, Gladine C, et al. Non-enzymatic cyclic oxygenated metabolites of adrenic, docosahexaenoic, eicosapentaenoic and alpha-linolenic acids; bioactivities and potential use as biomarkers[J]. Biochim Biophys Acta, 2015, 1851(4): 446-455. DOI: 10.1016/j.bbalip.2014.11.004.
2. Brahmbhatt V, Oliveira M, Briand M, et al. Protective effects of dietary EPA and DHA on ischemia-reperfusion-induced intestinal stress[J]. J Nutr Biochem, 2013, 24(1): 104-111. DOI: 10.1016/j.jnutbio.2012.02.014.
3. Kromhout D, Yasuda S, Geleijinse JM, et al. Fish oil and omega-3 fatty acids in cardiovascular disease: do they really work[J]. Eur Heart J, 2012, 33(4): 436-443. DOI: 10.1093/eurheartj/ehr362.
4. Nettleton JA, Katz R. n-3 long-chain polyunsaturated fatty acids in type 2 diabetes: a review[J]. J Am Diet Assoc, 2005, 105(3): 428-440. DOI: 10.1016/j.jada.2004.11.029.
5. Sterescu AE, Rousseau-Harsany E, farrell C, et al. The potential efficacy of omega-3 fatty acids as anti-angiogenic agents in benign vascular tumors of infancy[J]. Med Hypotheses, 2006, 66(6): 1121-1124. DOI: 10.1016/j.mehy.2005.12.040.
6. Serini S, Piccioni E, Calviello G. Dietary n-3 PUFA vascular targeting and the prevention of tumor growth and age-related macular degeneration[J]. Curr Med Chem, 2009, 16(34): 4511-4526.
7. Soderberg M, Edlund C, Kristensson K, et al. Lipid compositions of different regions of the human brain during aging[J]. J Neurochem, 1990, 54(2): 415-423.
8. Soderberg M, Edlund C, Kristensson K, et al. Fatty acid composition of brain phospholipids in aging and in Alzheimer’s disease[J]. Lipids, 1991, 26(6): 421-425.
9. McGahon MK, Zhang X, Scholfield CN, et al. Selective downregulation of the BKbeta1 subunit in diabetic arteriolar myocytes[J]. Channels (Austin), 2007, 1(3): 141-143.
10. Peng G, Xu J, Liu R, et al. Isolation, culture and identification of pulmonary arterial smooth muscle cells from rat distal pulmonary arteries[J/OL]. Cytotechnology, 2017, 2017: E1[201703-21]. . DOI: 10.1007/s10616-017-0081-8. [published online ahead of print].
11. Saleh SN, Albert AP, Peppiatt-Wildman CM, et al. Diverse properties of store-operated TRPC channels activated by protein kinase C in vascular myocytes[J]. J Physiol, 2008, 586(10): 2463-2476. DOI: 10.1113/jphysiol.2008.152157.
12. Ghatta S, Nimmagadda D, Xu X, et al. Large-conductance, calcium-activated potassium channels: structural and functional implications[J]. Pharmacol Ther, 2006, 110(1): 103-116. DOI: 10.1016/j.pharmthera.2005.10.007.
13. Hou S, Heinemann S, Hoshi T. Modulation of BKCa channel gating by endogenous signaling molecules[J]. Physiology (Bethesda), 2009, 24: 26-35. DOI: 10.1152/physiol.00032.2008.
14. Torres YP, Granados ST, Latorre R. Pharmacological consequences of the coexpression of BK channel alpha and auxiliary beta subunits[J]. Front Physiol, 2014, 5: 383. DOI: 10.3389/fphys.2014.00383.
15. Dopico AM, Bukiya AN. Lipid regulation of BK channel function[J]. Front Physiol, 2014, 5: 312. DOI: 10.3389/fphys.2014.00312.
16. Aires V, Hichami A, Filomenko R, et al. Docosahexaenoic acid induces increases in [Ca2+]i via inositol 1, 4, 5-triphosphate production and activates protein kinase C gamma and-delta via phosphatidylserine binding site: implication in apoptosis in U937 cells[J]. Mol Pharmacol, 2007, 72(6): 1545-1556. DOI: 10.1124/mol.107.039792.

Chinese Journal of Ocular Fundus Diseases

Effects of docosahexenoic acid on large conductance Ca⁺-activated K⁺ channels in retinal smooth muscle cells

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Chinese Journal of Ocular Fundus Diseases

Effects of docosahexenoic acid on large conductance Ca+-activated K+ channels in retinal smooth muscle cells

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Effects of docosahexenoic acid on large conductance Ca⁺-activated K⁺ channels in retinal smooth muscle cells