原代培养家兔肾脏内髓集合管细胞单层H^+/K^+交换测定

Fluorescent measurements of H^+/K^+ exchange in primary cultured inner medullary collecting duct cell monolayers or rabbit kidney

目的建立 ＩＭＣＤ细胞单层 Ｈ＋ ／Ｋ＋交换的测定方法。方法采用ＢＣＥＣＦ／ＡＭ荧光探针法，测定不同浓度ＣＯ２培养的ＩＭＣＤ细胞Ｈ＋／Ｋ＋交换。结果呼酸组的酸负荷后［ｐＨ］；恢复率为０．０６２０±０．０１２，高于对照组（０．０５０４±０．００９，Ｐ＜０．０５）。呼碱组为０．０４７６±０．００７，与对照组相比差异无显著性（Ｐ＞０．０５）。结论荧光探针法测定原代培养家兔肾脏ＩＭＣＤ细胞Ｈ＋／Ｋ＋交换，具有操作简便、稳定可靠、经济安全等优点。

AIM To establish the fluorescent measurement technique for H+ /K+ exchange in primary cultured IMCD cell monolayers of rabbit kidney. METHODS To measure H+ /K+ exchange in the primary cultured IMCD cell monolayers under different CO2 concentrations by the BCECF/AM fluorescent probe. RESULTS [pH], recovery rate after acid loaded in the respiratory acidosis group was 0.0620 ± 0.012, and was significantly higher than that of the control group (0.0504 ± 0.009, P < 0.05); which in the respiratory alkalosis group was 0.0476 ± 0.007, there was no signifacant difference between ALG and CG (P >0.05). CONCLUSION The fluorescent measurements of H+ /K+ exchange in primary cultured IMCD cell monolayers of rabbit kidney has many benefits which are convenient manipulation, stable and credit result, economy and safety, this method should further be popularized.