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目的 探讨呼吸性酸碱失衡对家兔内髓集合管 (IMCD)细胞碳酸酐酶 (CA)活性及其mRNA表达的影响。方法 采用咪唑 Tris技术测定呼吸性酸碱失衡IMCD细胞的CA活性 ;分别用原位杂交和RT PCR检测这些细胞CAⅣmRNA的表达变化。结果 呼酸组的CA的T1 2 明显缩短 ,而呼碱组显著延长。原位杂交和RT PCR结果均显示呼酸显著上调CAⅣmRNA表达 ;呼碱下调CAⅣmRNA表达 ,但与对照组无显著差别。结论 呼酸显著影响家兔肾脏IMCD细胞CA活性及其mRNA表达 ;呼碱显著降低CA活性 ,而CAⅣ的mRNA表达无明显下调 ,其原因尚等进一步研究。
Objective To investigate the effects of respiratory acid-base imbalance on carbonic anhydrase (CA) activity and mRNA expression in rabbit medullary capillaries (IMCD). Methods The activity of CA in respiratory and acid-base imbalance IMCD cells was measured by imidazole Tris technique. The expression of CA Ⅳ mRNA in these cells was detected by in situ hybridization and RT-PCR, respectively. Results The acitivic group CA1 T1 2 significantly shortened, while the calline group significantly prolonged. In situ hybridization and RT-PCR results showed that Huachang significantly up-regulated the expression of CA Ⅳ mRNA, while down-regulated the expression of CA Ⅳ mRNA by Huacha, but no significant difference with the control group. Conclusion Huachen significantly affected CA activity and mRNA expression in IMCD cells of rabbits, and the alkaloids significantly reduced the activity of CA, while the mRNA expression of CA Ⅳ was not significantly down-regulated. The reason is still under further investigation.