Effects Of Ca～(2+) Channel Blockers On Store-operated Ca～(2+) Channel Currents Of Kupffer Cell After Hepatic Ischemia/reperfusion Injury In Rats

AIM: To establish a method in isolating SD rat Kupffer cells which settles forpatch-clamp recording techniques. Using patch-clamp techniques to study the effectsof hepatic ischemia/reperfusion (I/R) injury on store-operated calcium channelcurrents (I_SOC) in freshly isolated Kupffer cells of rat; and the effects of Ca²⁺ channelblockers, 2-Aminoethoxydiphenyl borate (2-APB), SK&F96365, Econazole andMiconazole, on I_SOC of Kupffer cells.METHODS: The rat model of hepatic ischemia and repefusion injury was established.The effects of 2-APB (20μmol/L, 40μmol/L, 60μmol/L, 80μmol/L, 100μmol/L),SK&96365 (5μmol/L, 10μmol/L, 20μmol/L, 40μmol/L, 50μmol/L), econazole(0.1μmol/L, 0.3μmol/L, 1μmol/L, 3μmol/L, 10μmol/L), miconazole (0.1μmol/L,0.3μmol/L, 1μmol/L, 3μmol/L, 10μmol/L) on I_SOC of Kupffer cells were investigatedby whole-cell patch-clamp technique.RESULTS: Kupffer cells were obtained with the purity of 85% and an averageviability of 85%. Ischemia/reperfusion injury significantly increased I_SOC of Kupffercells to -159.47±34.49pA from sham-operated control of -80.44±25.22 pA, with thestatistically difference (P＜0.01, n=30). 2-APB (20, 40, 60, 80, 100μmol/L),SK&F96365 (5, 10, 20, 40, 50μmol/L), Econazole (0.1, 0.3, 1, 3, 10μmol/L) andMiconazole (0.1, 0.3, 1, 3, 10μmol/L) induced a concentration-dependent decrease ofI_SOC with IC₅₀ value of 37.41μmol/L (n=8), 5.89μmol/L (n=11), 0.21μmol/L (n=13),0.28μmol/L (n=10).CONCLUSION: This experiment established a method in isolating SD rat Kupffercells which settled for patch-clamp recording techniques successfully. We confirmedthat the SD rat Kupffer cells plasma membrane possesses SOC. We established a model of hepatic ischemia/reperfusion which settled for patch-clamp recordingtechniques successfully, also isolated the Kupffer cells after the hepaticischemia/reperfusion for studing the properties of I_SOC in this situation. Hepaticischemia/reperfusion could increase Isoc of Kupffer cells, leading to its activation,giving rise to hepatocytes injury. Calcium channel blockers, 2-APB, SK&F96365,Econazole and Miconazole, had obvious protective effects on ischemia/reperfusioninjury, probably through inhibiting Isoc of Kupffer cells, so that the activation ofKupffer cells can be prevented.