Benishin C G, Labedz T, Guo D D, Lewanczuk R Z, Pang P K
Department of Physiology, Faculty of Medicine, University of Alberta, Edmonton, Canada.
Am J Hypertens. 1993 Feb;6(2):134-40. doi: 10.1093/ajh/6.2.134.
Parathyroid hypertensive factor (PHF) is a newly described hypertensive factor isolated from the plasma of spontaneously hypertensive rats (SHR). Recent studies have suggested that the primary origin of PHF is the parathyroid gland (PG). In the present investigation, PG from spontaneously hypertensive rats (SHR), as well as from normotensive rats, were isolated and maintained in culture. The PG from SHR, but not normotensive rats, released PHF into the culture medium. Omission of calcium from the culture medium stimulated the release of PHF. For purification of PHF, parathyroid gland culture medium (PGCM) was first dialyzed at 1000 mwco, and then ultrafiltered at 5000 molecular weight cut-off (mwco). PHF activity was retained in the fraction that was greater than 1000 daltons and less than 5000 daltons. Dialyzed and filtered SHR PGCM was fractionated by molecular exclusion HPLC. Biologically active PHF was collected in a discrete region. The biologically active molecular exclusion fraction was subsequently fractionated by reverse-phase HPLC (C-8). PHF was collected in a single discrete peak, which did not occur in culture medium prepared from normotensive PG in a similar manner. This biologically active peak occurred in the same position on molecular exclusion and reverse-phase HPLC as PHF purified from SHR plasma using similar procedures. Incubation of PGCM with trypsin inactivates the biological activity of PHF. The UV spectrum of PGCM PHF is identical to that obtained from purified plasma PHF. These results are consistent with the presence of a peptide moiety in PHF, and support the parathyroid origin of plasma PHF.
甲状旁腺高血压因子（PHF）是一种新发现的从自发性高血压大鼠（SHR）血浆中分离出来的高血压因子。最近的研究表明，PHF的主要来源是甲状旁腺（PG）。在本研究中，从自发性高血压大鼠以及正常血压大鼠中分离出甲状旁腺并进行培养。来自SHR的甲状旁腺而非正常血压大鼠的甲状旁腺，将PHF释放到培养基中。培养基中去除钙会刺激PHF的释放。为了纯化PHF，首先将甲状旁腺培养基（PGCM）在截留分子量为1000的条件下进行透析，然后在截留分子量为5000的条件下进行超滤。PHF活性保留在分子量大于1000道尔顿且小于5000道尔顿的组分中。经透析和过滤的SHR PGCM通过分子排阻高效液相色谱法进行分离。具有生物活性的PHF收集在一个离散区域。随后，将具有生物活性的分子排阻组分通过反相高效液相色谱法（C - 8）进行分离。PHF收集在一个单一的离散峰中，而以类似方式从正常血压PG制备的培养基中未出现此峰。这个具有生物活性的峰在分子排阻和反相高效液相色谱上的位置与使用类似程序从SHR血浆中纯化的PHF相同。用胰蛋白酶孵育PGCM会使PHF的生物活性失活。PGCM PHF的紫外光谱与从纯化血浆PHF获得的光谱相同。这些结果与PHF中存在肽部分一致，并支持血浆PHF的甲状旁腺来源。
