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Diazoxide ameliorates microcirculatory disturbances through PKC-dependent Pathway in I/R-injured rat cremaster muscles

  • Published:
Journal of Biomedical Science

Summary

Diazoxide is a selective mitochondria ATP-sensitive potassium (KATP) channel opener, which has been reported to preserve the microvascular integrity of ischemia-reperfusion (I/R)-injured tissues. Our study aimed to assess diazoxide’s effects on I/R-injured cremaster muscles and to further elucidate its underlying mechanisms. Male Sprague Dawley (SD) rats were randomized (n = 8 per group) into four groups: sham-operated control group, I/R group (4 h of pudic epigastic artery ischemia followed by 2 h of reperfusion), diazoxide + I/R group, and chelerythrine (PKC inhibitor)+diazoxide+I/R group. Microscopically, we observed that I/R markedly increased the number of rolling, adhering, and transmigrating leukocytes. I/R also markedly decreased the number of functional capillaries. Biochemically, we found that I/R significantly increased TNF-α, E-selectin,L-selectin and P-selectin expressions. However, I/R did not cause significant changes in ICAM-1 and PECAM-1 expressions. On the other hand, in I/R + diazoxide group, we found that diazoxide reduced the number of rolling, adhering, and transmigrating leukocytes. Furthermore, biochemical study revealed that diazoxide caused only a decrease in L-selectin expression but had no effect on TNF-α, E-selectin, P-selectin, ICAM-1, and PECAM-1 expressions. Finally, in chelerythrine + diazoxide + I/R group, we observed that diazoxide’s protective effects were blocked by the addition of chelerythrine. Diazoxide’s ability to protect against I/R injury was confirmed by the observation that it reduced the number of rolling, adhering, and transmigrating leukocytes, and increased the number of functional capillaries. Our results indicated that diazoxide operated via a PKC-dependent pathway to achieve protection against I/R injury.

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Abbreviations

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

ICAM-1:

intercellular adhesion molecule 1

IPC:

ischemic preconditioning

I/R:

ischemia reperfusion

KATP:

ATP-sensitive K+ channels

Mabs:

monoclonal antibodies

PECAM-1:

platelet-endothelial cell adhesion molecule-1

PKC:

protein kinase C

PMN:

polymorphonuclear

RT-PCR:

reverse transcription polymerase chain reaction

TNF-α:

tumor necrosis factor alpha

VCAM-1:

vascular cell adhesion molecule-1

References

  1. J.N. Crinnion S. Homer-Vanniasinkam M.J. Gough (1993) ArticleTitleSkeletal muscle reperfusion injury: pathophysiology and clinical considerations Cardiovasc. Surg. 1 317–324 Occurrence Handle8076053

    PubMed  Google Scholar 

  2. D.L. Carden D.N Granger (2000) ArticleTitlePathophysiology of ischaemia reperfusion injury J. Pathol. 190 255–266 Occurrence Handle10685060

    PubMed  Google Scholar 

  3. M. Suematsu F.A. DeLano D. Poole R.L. Engler M. Miyasaka B.W. Zweifach G.W. Schmid-Schonbein (1994) ArticleTitleSpatial and temporal correlation between leukocyte behavior and cell injury postischemic rat skeletal muscle microcirculation Lab. Invest. 70 684–695 Occurrence Handle7910874

    PubMed  Google Scholar 

  4. G.W. Schmid-Schönbein R. Skalak K. Sung et al. (1983) Human leukocytes in the active state U. Bagge G.V.R Born P. Gaehtgens (Eds) White Blood Cells, Morphology and Rheology as Related to Function Martinus Mijhoff The Hague 21–31

    Google Scholar 

  5. R. Rothlein (1997) ArticleTitleOverview of leukocyte adhesion Neurology 49 S3–S4 Occurrence Handle9371140

    PubMed  Google Scholar 

  6. T.M. Carlos J.M. Harlan (1994) ArticleTitleLeukocyte-endothelial adhesion molecules Blood 84 2068–2101 Occurrence Handle7522621

    PubMed  Google Scholar 

  7. W.S. Somers J. Tang G.D. Shaw R.T. Camphausen (2000) ArticleTitleInsights into the molecular basis of leukocyte tethering and rolling revealed by structures of P- and E-selectin bound to SLe(X) and PSGL-1 Cell 103 467–479 Occurrence Handle11081633

    PubMed  Google Scholar 

  8. R.P. McEver (1991) ArticleTitleSelectins: novel receptors that mediate leucocyte adhesion during inflammation Thromb. Haemost. 65 223–228 Occurrence Handle1710835

    PubMed  Google Scholar 

  9. I. Inoue H. Nagase K. Kishi T. Higuti (1991) ArticleTitleATP-sensitive K+ channels in the mitochondrial inner membrane Nature 352 244–247 Occurrence Handle1857420

    PubMed  Google Scholar 

  10. G.J. Gross J.A. Auchampach (1992) ArticleTitleBlockade of ATP-sensitive potassium channels prevents myocardial preconditioning in dogs Circ. Res. 70 223–233 Occurrence Handle1310443

    PubMed  Google Scholar 

  11. G.J. Gross R.M. Fryer (1999) ArticleTitleSarcolemmal versus mitochondrial ATP-sensitive K+ channels and myocardial preconditioning Circ. Res. 84 973–979 Occurrence Handle10325234

    PubMed  Google Scholar 

  12. T. Mizumura K. Nithipatikom G.J. Gross (1995) ArticleTitleBimakalim, an ATP-sensitive potassium channel opener, mimics the effect of ischemic preconditioning to reduce infarct size, adenosine release, and neutrophil function in the dogs Circulation 92 1236–1245 Occurrence Handle7648671

    PubMed  Google Scholar 

  13. K.D. Garlid P. Paucek V. Yarov-Yarovoy H.N. Murray R.B. Darbenzio A.J. D’Alonzo N.J. Lodge M.A. Smith G.J. Grover (1997) ArticleTitleCardioprotective effect of diazoxide and its interaction with mitochondrial ATP-sensitive K+ channels: possible mechanism of cardioprotection Circ. Res. 81 1072–1081 Occurrence Handle9400389

    PubMed  Google Scholar 

  14. H. Ito Y. Taniyama K. Iwakura N. Nishikawa T. Masuyama T. Kuzuya M. Hori Y. Higashino K. Fujii T. Minamino (1999) ArticleTitleIntravenous nicorandil can preserve microvascular integrity and myocardial viability in patients with reperfused anterior wall myocardial infarction J. Am. Coll. Cardiol. 33 654–660 Occurrence Handle10080465

    PubMed  Google Scholar 

  15. H. Liu H.Y. Zhang X. Zhu Z. Shao Z. Yao (2002) ArticleTitlePreconditioning blocks cardiocyte apoptosis: role of K (ATP) channels and PKC-epsilon Am. J. Physiol. Heart Circ. Physiol. 282 H1380–H1386 Occurrence Handle11893574

    PubMed  Google Scholar 

  16. Y. Wang E. Takashi M. Xu A. Ayub M. Ashraf (2001) ArticleTitleDownregulation of protein kinase C inhibits activation of mitochondrial K (ATP) channels by diazoxide Circulation 104 85–90 Occurrence Handle11435343

    PubMed  Google Scholar 

  17. S. Baez (1973) ArticleTitleAn open cremaster muscle preparation for the study of blood vessels by in vivo microscopy Microvasc. Res. 5 384–394 Occurrence Handle4709735

    PubMed  Google Scholar 

  18. A. Noma (1983) ArticleTitleATP-regulated K1 channels in cardiac muscle Nature 305 147–148 Occurrence Handle6310409

    PubMed  Google Scholar 

  19. P. Rorsman K. Bokvist C. Ammala P. Arkhammar P.O. Berggren O. Larsson K. Wahlander (1991) ArticleTitleActivation by adrenaline of a low-conductance G protein-dependent K+ channel in mouse pancreatic B cells Nature 349 77–79 Occurrence Handle1898674

    PubMed  Google Scholar 

  20. U. Russ F. Metzger E. Kickenweiz A. Hambrock P. Krippeit-Drews U. Quast (1997) ArticleTitleBinding and effects of KATP channel openers in the vascular smooth muscle cell line, A10 Br. J. Pharmacol. 122 1119–1126 Occurrence Handle9401776

    PubMed  Google Scholar 

  21. J.J. Nielsen M. Kristensen Y. Hellsten J. Bangsbo C. Juel (2003) ArticleTitleLocalization and function of ATP-sensitive potassium channels in human skeletal muscle Am. J. Physiol. Regul. Integr. Comp. Physiol. 284 R558–R563 Occurrence Handle12388475

    PubMed  Google Scholar 

  22. C. Jiang G.G. Haddad (1997) ArticleTitleModulation of K+ channels by intracellular ATP in human neocortical neurons J. Neurophysiol. 77 93–102 Occurrence Handle9120601

    PubMed  Google Scholar 

  23. G.J. Grover K.D. Garlid (2000) ArticleTitleATP-sensitive potassium channels: a review of their cardioprotective pharmacology J. Mol. Cell. Cardiol. 32 677–695 Occurrence Handle10756123

    PubMed  Google Scholar 

  24. F. Chamoun M. Burne M. O’Donnell H. Rabb (2000) ArticleTitlePathophysiologic role of selectins and their ligands in ischemia reperfusion injury Front. Biosci. 5 E103–E109 Occurrence Handle11056080

    PubMed  Google Scholar 

  25. C.D. Collard S. Gelman (2001) ArticleTitlePathophysiology, clinical manifestations, and prevention of ischemia-reperfusion injury Anesthesiology 94 1133–1138 Occurrence Handle11465607

    PubMed  Google Scholar 

  26. G.J. del Zoppo G.W. Schmid-Schonbein E. Mori B.R. Copeland C.M. Chang (1991) ArticleTitlePolymorphonuclear leukocytes occlude capillaries following middle cerebral artery occlusion and reperfusion in baboons Stroke 22 1276–1283 Occurrence Handle1926239

    PubMed  Google Scholar 

  27. J. Bjork P. Hedqvist K.E. Arfors (1982) ArticleTitleIncrease in vascular permeability induced by leukotriene B4 and the role of polymorphonuclear leukocytes Inflammation 6 189–200 Occurrence Handle6179872

    PubMed  Google Scholar 

  28. U.H. Von Andrian J.D. Chambers L.M. McEvoy R.F. Bargatze K.E. Arfors E.C. Butcher (1991) ArticleTitleTwo-step model of leukocyte-endothelial interaction in inflammation: distinct roles for LECAM-1 and the leukocyte β2 integrins in vivo Proc. Natl. Acad. Sci. USA 88 7538–7542 Occurrence Handle1715568

    PubMed  Google Scholar 

  29. W.J. Sanders E.J. Gordon O. Dwir P.J. Beck R. Alon L.L. Kiessling (1999) ArticleTitleInhibition of L-selectin-mediated rolling by synthetic glycoprotein mimics J. Biol. Chem. 274 5271–5278 Occurrence Handle10026133

    PubMed  Google Scholar 

  30. B. Haribabu D.A. Steeber H. Ali R.M. Richardson R. Snyderman T.F. Tedder (1997) ArticleTitleChemoattractant receptor-induced phosphorylation of L-selectin J. Biol. Chem. 272 13961–13965 Occurrence Handle9153259

    PubMed  Google Scholar 

  31. C. Feehan K. Darlak J. Kahn B. Walcheck A.F. Spatola T.K. Kishimoto (1996) ArticleTitleShedding of lymphocyte L-selectin adhesion molecule is inhibited by a hydroxamic acid-based protease inhibitor J. Biol. Chem. 271 7019–7024 Occurrence Handle8636132

    PubMed  Google Scholar 

  32. J. Kahn B. Walcheck G.I. Migaki M.A. Jutila T.K. Kishimoto (1998) ArticleTitleCalmodulin regulates L-selectin adhesion molecule expression and function through a protease-dependent mechanism Cell 92 809–818 Occurrence Handle9529256

    PubMed  Google Scholar 

  33. E. Matala S.R. Alexander T.K. Kishimoto B. Walcheck (2001) ArticleTitleThe cytoplasmatic domain of L-selectin participates in regulating L-selectin endoproteolysis J. Immunol. 167 1617–1623 Occurrence Handle11466384

    PubMed  Google Scholar 

  34. O. Spertini G.S. Kansas J.M. Munro J.D. Griffin T.F. Tedder (1991) ArticleTitleRegulation of leukocyte migration by activation of the leukocyte adhesion molecule-1 (LAM-1) selectin Nature 349 691–694 Occurrence Handle1705015

    PubMed  Google Scholar 

  35. J.R. Parratt K.A. Kane (1994) ArticleTitleKATP channels and ischaemic preconditioning Cardiovasc. Res. 28 783–787 Occurrence Handle7923280

    PubMed  Google Scholar 

  36. M.E. Speechly-Dick M.M. Mocanu D.M. Yellon (1994) ArticleTitleProtein kinase C: its role in ischemic preconditioning in the rat Circ. Res. 75 586–590 Occurrence Handle8062429

    PubMed  Google Scholar 

  37. M.E. Speechly-Dick G.J. Grover D.M. Yellon (1995) ArticleTitleDoes ischemic preconditioning in the human involve protein kinase C and the ATP-dependent K+ channel?: studies of contractile function after simulated ischemia in an atrial in vitro model Circ. Res. 77 1030–1035 Occurrence Handle7554138

    PubMed  Google Scholar 

  38. D.P. Jenkins M. Kerac D.M. Yellon (1995) ArticleTitlePreconditioning: protein kinase C and the KATP channel in isolated rat heart J. Mol. Cell. Cardiol. 27 A154

    Google Scholar 

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Authors and Affiliations

  1. Department of Life Science and Plastic & Reconstructive Surgery, College of Medicine, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan, Taiwan

    William Wei, Fu-Chan Wei & Li-Man Hung

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  1. William Wei
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  2. Fu-Chan Wei
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Correspondence to Li-Man Hung.

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Wei, W., Wei, FC. & Hung, LM. Diazoxide ameliorates microcirculatory disturbances through PKC-dependent Pathway in I/R-injured rat cremaster muscles. J Biomed Sci 12, 521–529 (2005). https://doi.org/10.1007/s11373-005-3730-4

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