저칼륨혈증 흰쥐 신장에서 Carbonic anhydrase II 발현 증가
Enhanced Expression of Carbonic anhydrase II in Hypokalemic Rat Kidney
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초록
산-염기 평형장애나 전해질 불균형은 신요세관에서 HCO_3^- 재흡수가 증가되거나 감소되는 것과 연관이있다고 알려져 있다. 칼륨제한시 대사성 알칼리증을 유발시키고 이와 관련된 Na/H exchanger와 Na/HCO_3 cotransporter에대한 연구는 잘 알려져 있지만 저칼륨혈증시 carbonic anhydrase에 관한 연구는 없다. 본 연구는 흰쥐신장에서 칼륨제한 식이 시기에 따라 carbonic anhydrase II의 신장내 발현 및 분포의 변화를 Western 분석과 면역조직화학적 방법으로 관찰하였다. Western 분석 소견에서 carbonic anhydrase II 단백은 30 kDa 정도이고 정상식이 군에서 상당량 발현되었으며칼륨제한 식이 군에서는 정상식이 군에 비해 약간 증가하는 경향이었다. 면역조직화학 소견에서 carbonic anhydrase II 단백은 겉질집합관, 바깥속질집합관 및 속속질집합관의 상 1/3에서만 발현되었다. 면역반응성은 속속질집합관으로 갈수록 감소하였다. 집합관에서의 면역반응성은 개재세포가 강했으며 주세포는 미약하였다. 칼륨제한 식이 군에서 면역반응 부위는 정상식이 군과 차이가 없었다. 면역반응성은 겉질집합관 특히 일부 개재세포와 주세포에서 감소하였고 바깥속질 바깥줄무늬층 집합관에서는 변화가 없었다. 이와는 대조적으로 바깥속질속줄무늬층 집합관과 속속질집합관 상 1/3에서는 현저히 증가하였다. 이상의 소견은 저칼륨혈증시 신장 집합관에서의 carbonic anhydrase II 단백은 부위에 따라 발현양상이 다름을암시하였다.
Abstract
A number of acid-base or electrolyte disorders are associated with decreased or increased HCO_3^-reabsorption in the renal tubules. The present study was to examine the alterations of expression and distribution of Carbonic anhydrase II in the kidneys of normal and potassium-depleted rats using Western blot analysis and immuno-histochemistry. Western blot analysis demonstrated that CA II protein, ~30 kDa at molecular mass, was abundantly expressed in normal group. All potassium-depleted groups showed slightly increased CA II protein compared to normal group. In control group, immunoreactivity of CA II protein was detected in the entire collecting duct. Signal intensity was prominent in the intercalated cells and weak in the principal cells of the cortical collecting ducts. In potassiumdepleted groups, the pattern of cellular labeling of CA II protein was identical to that of normal group, but the signal intensity was decreased in cortical collecting duct, markedly increased in the inner stripe of outer medullary and inner medullary collecting ducts, and unchanged in the outer stripe of outer medullary collecting duct. These results suggest that chronic hypokalemia impact the expression pattern of CA II protein depending the portion of the collecting duct.
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