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首页> 外文期刊>Cellular Physiology and Biochemistry >Regulation of intracellular pH by bovine intervertebral disc cells
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Regulation of intracellular pH by bovine intervertebral disc cells

机译：牛椎间盘细胞对细胞内pH的调节

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Extracellular acidity is an important determinant of intervertebral disc matrix turnover, possibly exerting effects through changes of intracellular pH (pH(i)). There is, however, little information concerning the ways in which these cells regulate their pH(i), Fluorimetric techniques have been used in the present study to measure pH(i) in isolated intervertebral disc cells, and to characterise the membrane transport pathways by which it is regulated. Nucleus pulposus cells were obtained from bovine intervertebral discs by standard enzymatic digestion techniques, and loaded with the pH-sensitive fluoroprobe BCECF. Resting pH(i) was approximately 6.7 for cells suspended in either HEPES-buffered (HBS) or CO2/HCO3--buffered (BBS) media. Intrinsic buffering capacity was approximately 19 mM pH unit(-1) in HBS and was increased when cells were suspended in BBS. A combination of ion substitution and inhibitor studies for cells at steady-state pH. or acidified by exposure to NH4Cl revealed that in HBS Na+ x H+ exchange and an H+-ATPase extrude acid from these cells. Only one of these two systems, the Na+ x H+ exchanger, exhibited a sensitivity to pH(i), identifying it as the regulator of pH(i) under these conditions. In BBS, an additional pathway which was dependent on extracellular Na+, extracellular HCO3- and intracellular Cl- was detected. These properties are consistent with the four ion HCO3--dependent transporter, although the cation-rich, anion-poor extracellular matrix of the intervertebral disc means that such a pathway has only a marginal role in disc cell pH(i) regulation. Copyright (C) 2000 S. Karger AG, Basel. [References: 27]
机译：细胞外酸度是椎间盘基质周转的重要决定因素，可能通过改变细胞内pH（pH（i））发挥作用。但是，关于这些细胞调节其pH（i）的方式的信息很少。在本研究中，荧光技术已用于测量分离的椎间盘细胞中的pH（i），并通过以下方法表征膜转运途径它受到监管。通过标准的酶消化技术从牛椎间盘获得髓核细胞，并装载pH敏感的氟探针BCECF。对于悬浮在HEPES缓冲（HBS）或CO2 / HCO3缓冲（BBS）培养基中的细胞，静止pH（i）约为6.7。 HBS中的固有缓冲能力约为19 mM pH单位（-1），当细胞悬浮于BBS中时，固有缓冲能力会增加。在稳态pH下对细胞进行离子取代和抑制剂研究相结合。或通过暴露于NH4Cl进行酸化显示，在HBS中，Na + x H +交换和H + -ATPase从这些细胞中挤出酸。这两个系统中只有一个，即Na + x H +交换剂，对pH（i）表现出敏感性，从而将其确定为这些条件下pH（i）的调节剂。在BBS中，检测到另一条依赖于细胞外Na +，细胞外HCO3-和细胞内Cl-的途径。这些特性与四离子HCO3依赖性转运蛋白一致，尽管椎间盘的富含阳离子，贫于阴离子的细胞外基质意味着这种途径在椎间盘细胞pH（i）调节中仅具有边际作用。版权所有（C）2000 S.Karger AG，巴塞尔。 [参考：27]

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