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目的探讨血浆钙水平对肾组织斯钙素-1(STC1)基因表达与质膜Ca2+-ATP酶(PMCA)活性的影响。方法将72只成体中华大蟾蜍随机分为3组,分别暴露于添加0.1mol/L CaCl2的自来水、添加0.03mol/L乙二胺四乙酸(EDTA)的自来水及自来水中,于暴露前及暴露后12、24、48、72、96、120和144h取血浆和肾脏,利用比色法和半定量RT-PCR法分别检测血浆钙水平、PMCA活性和STC1 mRNA水平,利用免疫组织化学染色确定STC1免疫反应。结果 0.1mol/L CaCl2暴露致中华大蟾蜍血浆钙水平在12和24h时升高,48h时降至正常水平,48h后持续升高;12～48h内肾组织PMCA活性增强,STC1 mRNA水平上调,72h后回复至正常水平。而0.03mol/L EDTA暴露则引起血浆钙水平下降,但肾组织PMCA活性和STC1 mRNA水平与对照组相比未出现明显变化。免疫组织化学技术显示,肾脏远曲小管和集合管上皮细胞出现STC1免疫反应,且0.1mol/L CaCl2暴露可致STC1免疫反应增强。结论血浆钙水平升高致肾小管和集合管上皮细胞PMCA活性增强,STC1表达上调,而STC1表达上调又使血浆钙水平下降;但血浆钙水平持续升高则使PMCA活性下降,STC1基因表达下调。
Objective To investigate the effect of plasma calcium on the expression of stanamycin-1 (STC1) gene and the activity of plasma membrane Ca2 + -ATPase (PMCA) in kidney. Methods Seventy-two adult Bufo bufo gargarizans were randomly divided into three groups. The animals were exposed to tap water supplemented with 0.1 mol / L CaCl2, tap water and tap water supplemented with 0.03 mol / L ethylenediaminetetraacetic acid (EDTA) The plasma and kidney were taken at 12, 24, 48, 72, 96, 120 and 144 h after operation. The plasma calcium level, PMCA activity and STC1 mRNA level were detected by colorimetric assay and semi-quantitative RT-PCR. STC1 immune response. Results The plasma calcium level of Bufo bufo gargarizans exposed to 0.1 mol / L CaCl2 increased at 12 and 24 h, decreased to normal level at 48 h, and continued to increase at 48 h. The activity of PMCA increased and the level of STC1 mRNA increased in 12 ~ 48 h, After 72h returned to normal levels. However, 0.03mol / L EDTA caused a decrease in plasma calcium levels, but there was no significant change in PMCA activity and STC1 mRNA level in the kidney. Immunohistochemistry showed that STC1 immunoreactivity was found in the distal convoluted tubules and collecting duct epithelial cells of the kidney, and the exposure of 0.1 mol / L CaCl2 resulted in the enhancement of STC1 immune response. Conclusions The increase of plasma calcium level leads to the enhancement of PMCA activity in renal tubular and collecting duct epithelial cells, the upregulation of STC1 expression and the upregulation of STC1, which in turn leads to the decrease of plasma calcium level. However, the increase of plasma calcium level decreases PMCA activity and STC1 gene expression .