目的 探索三甲基氯化锡(trimethyltin chloride,TMT)对亚慢性染毒大鼠肾氢钾ATP酶活力、蛋白和基因表达水平的影响.方法 取6周龄雌性SD大鼠55只,随机分成对照、低剂量、中剂量、高剂量和超高剂量组,共5组,每组11只,分别按0、8.20、32.81、131.25、262.50μg·kg-1·d-1的TMT剂量经饮水亚慢性染毒,14周后测定血钾水平,取大鼠肾脏,用定磷法测量氢钾ATP活力,用免疫印迹法测定肾氢钾ATP酶蛋白表达水平,用实时定量-聚合酶链反应法测定肾氢钾ATP酶的mRNA表达水平.结果 与对照组[(6.9±0.3)mmol/L]比较,超高剂量组血钾水平[(5.6±0.4)mmol/L]明显降低,差异有统计学意义(P<.01).低剂量和超高剂量组氢钾ATP酶活力分别为(4.50±1.45)、(435±-0.72)μmolPi·mgprot-1·h-1,明显低于对照组[(6.55±0.77)μmolPi·mgprot-1·h-1],差异有统计学意义(P<0.05).与对照组比较,超高剂量组氢钾ATP酶蛋白和mRNA表达水平差异均无统计学意义(P>0.05).结论 TMT亚慢性接触可抑制氢钾ATP酶的活力,但不抑制该酶的蛋白和基冈表达.
Objective To study the activity、protein and gene expression of renal HK-ATPase (HKA) in rats subchronicly exposed to trimethyltin chloride (TMT). Methods In subchronic toxic test (14-week), 55 female SD rats (age, 6 weeks) were divided randomly into 5 groups: control, low, medium, high and super high dosage, respectively, which drank water with TMT of 0, 8.20, 32.81, 131.25 and 262.50 μg·kg-1 ·d-1 for 14 weeks.Then serum K+ levels were measured; the activities of H K-ATPase (HKA) in kidneys were detected by the method of determinenate phosphorus content;Western Blot assay and real-time PCR were used to exam the protein and mRNA expression levels of HKA in kidneys, respectively. Results The serum K + level in super-high dosage group was (5.6±0.4) mmol/L, which was significantly lower than that [(6.9±0.3) mmol/L] in control group (P<0.01).The HKA enzymatic activity of kidneys in low and super high dosage groups was 4.50±1.45 and 4.55±0.72 μmolPi·mg prot-1h-1, respectively, which were significantly lower than that (6.55±0.77 μmol Pi ·mg prot-1h-1) in control group (P<0.05). Conclusion When rats were exposed subchronicly to TMT, the renal HKA activity could reduce, but the expression levels of HKA protein and mRNA did not decrease.