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目的 研究肾脏内髓集合管(IMCD)细胞Cl~-/HCO_3~_交换在呼吸性酸碱失衡时的代偿调节作用。方法 分别以3％CO_2和10％CO_2模拟慢性呼吸性碱中毒(呼碱,ALG组)和慢性呼吸性酸中毒(呼酸,ACG组)建立家兔肾脏IMCD细胞培养模型,对照组(CG组)为5％CO_2+空气平衡,采用荧光探针法测定IMCD细胞Cl~-/HCO_3~-交换,用原位杂交和逆转录-聚合酶链反应(RT-PCR)检测IMCD细胞肾脏阴离子交换蛋白2(KAE2)mRNA表达。结果 ACG组Cl~-/HCO_3~-交换[(0.132±0.015)pHi/min]显著高于CG组[(0.117±0.016)pHi/min,P<0.05]CG组与ALG组[(0.111±0.015)pHi/min]无显著差别(P>0.05)。ACG组KAE2 mRNA原位杂交积分光密度(325.48±90.84)显著高于CG组(68.54±20.28,P<0.01);CG组与ALG组(50.56±16.08)无显著差别(P>0.05)。ACG组KAE2 mRNA的RT-PCR积分光密度比值(2.24±0.36)显著高于CG组(0.43±0.05,P<0.01);CG组与ALG组(0.37±0.05)无显著差别(P>0.05)。结论 慢性呼酸显著增加IMCD细胞Cl~-/HCO_3~-交换和KAE2 mRNA表达,慢性呼碱略降低IMCD细胞Cl~-/HCO_3~-交换和KAE2 mRNA表达。
Objective To study the compensatory regulation of Cl ~ - / HCO_3 ~ _ exchange in renal medullary collecting duct (IMCD) cells in respiratory acid-base imbalance. Methods Rabbit kidney IMCD cell culture model was established by simulating chronic respiratory alkalosis (aldosterone, ALG group) and chronic respiratory acidosis (aconite, ACG group) with 3% CO 2 and 10% CO 2 respectively. The control group (CG group ) Was 5% CO 2 + air balance. The Cl ~ - / HCO 3 ~ - exchange of IMCD cells was detected by fluorescent probe method. The in situ hybridization and reverse transcription - polymerase chain reaction (RT - PCR) (KAE2) mRNA expression. Results The Cl ~ - / HCO_3 ~ exchange [(0.132 ± 0.015) pHi / min] in ACG group was significantly higher than that in CG group [(0.117 ± 0.016) pHi / min, P < ) pHi / min] no significant difference (P> 0.05). The integrated optical density of in situ hybridization of KAE2 mRNA in ACG group (325.48 ± 90.84) was significantly higher than CG group (68.54 ± 20.28, P <0.01). There was no significant difference between ACG group and ALG group (50.56 ± 16.08) (P> 0.05). The ratio of RT-PCR integrated optical density of KAE2 mRNA in ACG group (2.24 ± 0.36) was significantly higher than that in CG group (0.43 ± 0.05, P <0.01). There was no significant difference between CG group and ALG group (0.37 ± 0.05) . Conclusion Chronic acidosis can significantly increase Cl ~ - / HCO_3 ~ - exchange and KAE2 mRNA expression in IMCD cells. Chronic exhalation reduces Cl ~ - / HCO_3 ~ - exchange and KAE2 mRNA expression in IMCD cells slightly.