内毒素对体外培养仓鼠肾细胞膜酶PLA_2和ATPase活性影响及阳离子A拮抗效应研究

Studies on Endotoxin-induced Membrane Injury of Hamsters Rat Renal Cells in vitro and Protection of Cation A

为研究内毒素(Endotoxin,ET)对体外培养的仓鼠肾细胞膜活性影响及阳离子A(Cation A,CA)对其的保护效应。随机将体外培养的仓鼠肾细胞(BHK-21)分为:对照组(Ⅰ组)、ET处理组(Ⅱ组)和ET+CA组(Ⅲ组),采用微量滴定法和定磷法,第3、6、12、24小时测定细胞膜上磷脂酶A2(PLA2)和ATP酶(ATPase)的活性。结果表明,II组中PLA2活性在整个实验过程中均显著高于I组(P<0.01或P<0.05),Na+-K+-ATPase、Ca2+-Mg2+-ATPase活性呈现先增高后降低的趋势(3、6、12 h增高,24 h以后降低)。III组与II组相比,PLA2的活性显著降低(P<0.05),Na+-K+-ATPase、Ca2+-Mg2+-ATPase活性在整个实验过程中均显著增高(P<0.01或P<0.05)。细胞膜上的PLA2活性增高,ATPase的活性先增高后降低,提示细胞膜受到损伤;CA能明显改善内毒素所致的体外细胞膜损伤。

The experiment aimed at investigating the effect of endotoxin-induced on phospholipase A2（PLA2） and ATPase activities of Hamsters Rat Renal cells membrane injury in vitro.The cells were randomly divided into three groups： control group（groupⅠ）,endotoxin group（groupⅡ）,ET＋CA group（group Ⅲ）.Using trace titration and phosphate determination method,the activities of phospholipase A2（PLA2） and ATPase were detected in 3,6,12,24 h.Compared with the groupⅠ,the PLA2 in renal cell membrane of group Ⅱhad significantly higher activities at all time of the experiment（P0.01,P0.05）.The ATPase activities of cell membrane in group Ⅱincreased in 3,6,12 h and decreased significantly after 24 h.Compared with the group Ⅱ, the alteration of PLA2 activities of group Ⅲ decreased significantly（P0.01,P0.05）,and the ATPase activities of renal cell membrane of group Ⅲ increased or decreased significantly（P0.01,P0.05）.These results indicated that ET treatment increased PLA2 and ATPase activities in the cell membrane in the primary stage,while inhibited the ATPase activities after 24 h.Consequently,ET induced the cell membrane injury,whereas CA had prominent protective effects on renal cell membrane.