培养人眼小梁细胞水通道蛋白1与Na^+-K^+-ATP酶、Ca^(2+)-ATP酶的相关研究

Investigation of the Relationship between Aquaporin-1 with Na^+-K^+-ATPase and Ca^(2+)-ATPase in Cultured Human Trabecular Meshwork Cells

目的探讨人眼小梁细胞上水通道蛋白1(aquaporin-1,AQP1)与Na+-K+-ATP酶、Ca2+-ATP酶的相关关系。方法培养的人眼小梁细胞分别给予地塞米松(dexamethasone,DEX)0.4μg/mL(B组)、DEX0.4μg/mL+AQP1反义寡核苷酸(antisense oligonucleotides,AS-ODN)0.625μg/mL(C组)、DEX0.4μg/mL+AS-ODN 1.25μg/mL(D组)、DEX0.4μg/mL+AS-ODN 2.5μg/mL(E组)、DEX0.4μg/mL+AS-ODN 5μg/mL(F组)处理,以不加药组为对照组(A组)。5d后测定各组Na+-K+-ATP酶、Ca2+-ATP酶活性。结果与对照组的相比,DEX0.4μg/mL组Na+-K+-ATP酶活性、Ca2+-ATP酶活性明显下降,差异有统计学意义(P<0.05)。在各DEX+AS-ODN组,随着AS-ODN浓度加大,小梁细胞Na+-K+-ATP酶的活性逐渐提升。在AS-ODN浓度加到5μg/mL时,Na+-K+-ATP酶的活性恢复到最大。B、C、D、E组Na+-K+-ATP酶的活性与对照组及F组差异均有统计学意义(P<0.05),F组与对照组差异无统计学意义(P>0.05)。B、C、D、E、F组Ca2+-ATP酶活性与对照组差异均有统计学意义(P<0.05),B、C、D、E、F组之间Ca2+-ATP酶活性差异无统计学意义(P>0.05)。结论人眼小梁细胞AQP1与Na+-K+-ATP酶关系密切,与Ca2+-ATP酶关系不显著,AQP1的正常表达是Na+-K+-ATP酶活性维持的必要条件之一。

Objective To investigation of the relationship between aquaporin-1 （ AQP1 ） with Na ＋-K ＋-ATPase and Ca 2＋-ATPase in cultured human trabecular meshwork（ HTM） cells. Methods Cultured HTM cells in vitro were respectively trea-ted with 0. 4μg/mLdexamethasone（ DEX） ,0. 4μg/mLDEX and 0. 625μg/mLAQP1 antisense oligonucleotides（ AS-ODN） ,0. 4μg/mLDEX and 1. 25μg/mLAS-ODN, 0. 4μg/mLDEX and 2. 5μg/mLAS-ODN、0. 4μg/mLDEX and 5μg/mLAS-ODN. No drug group was set as the control. After 5d,use detecting pi to measure the activity of Na ＋-K ＋-ATPase and Ca 2＋-ATPase. Results In 0. 4μg/mLDEX group,the activity of Na ＋-K ＋-ATPase and Ca2＋-ATPase were less than control group（P〈0. 05）. In DEX and AS-ODN groups,the activity of Na ＋-K ＋-ATPase was reversible gradually. At 5μg/mLAS-ODN,the reverse of Na ＋-K ＋-ATPase activity reached maximum. There are a statistical difference between Na ＋-K ＋-ATPase activity of B、C、D、E groups with control group and F group. There are not a statistical difference between Na ＋-K ＋-ATPase activity of control group and F group. There are a statistical difference between Ca 2＋-ATPase activity of control group and B、C、D、E 、F groups. There are not a statistical difference among Ca 2＋-ATPase of B、C、D、E 、F groups. Conclusion The relationship AQP1 and Na ＋-K ＋-ATPase in cultured HTM cells is closed. The relationship AQP1 and Ca 2＋-ATPase in cultured HTM cells is not obvious. The normal expression of AQP1 is necessary to maintain the activity of Na ＋-K ＋-ATPase.