烫伤血清对大鼠库普弗细胞胞浆游离钙浓度的影响

The effect of postburn serum on the free intracellular calcium concentration of the in vitro cultured Kupffer cells

目的：观察烫伤血清对大鼠分离培养的库普弗细胞（ＫＣ）胞浆内游离钙（［Ｃａ２＋］ｉ）浓度的影响。方法：胶原酶灌注消化、梯度离心分离ＫＣ，在ＫＣ悬液中加入烫伤血清，Ｆｕｒａ－２／ＡＭ荧光染色法测定［Ｃａ２＋］ｉ。结果：分离培养的ＫＣ静息状态下［Ｃａ２＋］ｉ为（１０９．２９±４．１７）ｎｍｏｌ／Ｌ，烫伤血清可以使［Ｃａ２＋］ｉ显著升高，以烫伤后６ｈ的血清作用最明显；［Ｃａ２＋］ｉ的浓度随血清浓度的升高而升高；烫伤血清作用１ｍｉｎＫＣ［Ｃａ２＋］ｉ已明显升高，２ｍｉｎ达高峰，２０ｍｉｎ时仍明显高于假烫组。结论：烫伤血清能明显升高ＫＣ［Ｃａ２＋］ｉ，作用快速而持久。

Objective: To investigate the effect of stimulation of postburn serum on the free intracellular calcium concentration([ Ca^(2+)]i) of the in vitro cultured Kupffer cells(KC). Methods: KC was isolated by collagenase I using portal perfusion and centrifugation; [ Ca^(2+)]i was measured using the fluorescent dye Fura2/AM. Results: The resting [ Ca^(2+)]i was (109.29±4.17) nmol/L. Postburn serum could significantly increase [Ca^(2+)]i. The role of 6 h postburn serum was greatest. The higher concentration of postburn serum, the more increase in [ Ca^(2+)]i. One minute′ action of postburn serum on KC increased [ Ca^(2+)]i. Significant increase could still be seen after twenty minutes′ action. Conclusion: Postburn serum can significantly increase [ Ca^(2+)]i of in vitro KC. Its effect is quick,durable and concentration dependent.