Cheleuitte D, Mizuno S, Glowacki J
Department of Orthopedic Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
J Clin Endocrinol Metab. 1998 Jun;83(6):2043-51. doi: 10.1210/jcem.83.6.4848.
It has been proposed that cytokines mediate the acceleration of bone loss following menopause. Because of the intimate relationship between bone marrow stromal cells and bone tissue, it is possible that marrow cells and their products contribute to the bone microenvironment and influence the regulation of bone cell differentiation and activity. We examined the production of cytokines by bone marrow stromal cells from a total of 37 women and 15 men undergoing total hip replacement for noninflammatory joint disease. Low-density mononuclear cells were isolated from bone marrow and were cultured in phenol red-free alpha MEM medium supplemented with 10% FBS and antibiotics. Constitutive secretion of interleukin-6 (IL-6) was positively correlated with age in a series of 8 women and 5 men measured by bioassay (r = 0.98; P < 0.01) and in a series of 18 women and 10 men measured by immunoassay (r = 0.56; P < 0.01). The pattern of cytokine production by bone marrow stromal cells was examined in detail in 23 postmenopausal women, aged 49-88 yr. Basal secretion of immunoreactive IL-6 and IL-11, but not granulocyte-macrophage colony-stimulating factor, increased with time in culture. Exogenous IL-1 beta stimulated secretion of IL-6 and IL-11 in a saturable, dose-dependent manner. Secretion of soluble IL-6 receptor was not correlated with secretion of IL-6, either constitutively or in the presence of IL-1 beta. In 4 of 14 samples, IL-1 beta also stimulated secretion of granulocyte-macrophage colony-stimulating factor. IL-1 beta was undetectable in 7 of 9 cultures during the 2-week culture period. IL-6 did not stimulate secretion of IL-1 beta in the 7 cultures tested. Cells were dependent upon serum for viability and growth and were not sustained by a serum substitute (1% insulin-transferrin-selenium-BSA). Cells grown in medium with 10% FBS and supplemented with 1% insulin-transferrin-selenium-BSA secreted 10-fold more IL-6 than cells grown in serum alone. Marrow from 7 women receiving estrogen replacement therapy showed lower constitutive secretion of IL-6 (75%; P < 0.006) and IL-11 (43%; P < 0.05) than marrow from age-matched controls and had blunted stimulation of IL-6 and IL-11 secretion by exogenous IL-1 beta. These data indicate distinct patterns of cytokine production by human marrow stromal cultures dependent upon age and estrogen status.
有人提出，细胞因子介导绝经后骨质流失的加速。由于骨髓基质细胞与骨组织之间关系密切，骨髓细胞及其产物有可能对骨微环境产生影响，并影响骨细胞分化和活性的调节。我们检测了37名女性和15名男性因非炎性关节疾病接受全髋关节置换术患者的骨髓基质细胞产生细胞因子的情况。从骨髓中分离出低密度单核细胞，并在添加10%胎牛血清和抗生素的无酚红α-MEM培养基中培养。通过生物测定法对8名女性和5名男性进行检测，白细胞介素-6（IL-6）的组成性分泌与年龄呈正相关（r = 0.98；P < 0.01）；通过免疫测定法对18名女性和10名男性进行检测，也得到类似结果（r = 0.56；P < 0.01）。我们详细检测了23名年龄在49 - 88岁的绝经后女性骨髓基质细胞产生细胞因子的模式。免疫反应性IL-6和IL-11的基础分泌量，但不包括粒细胞-巨噬细胞集落刺激因子，随着培养时间增加。外源性IL-1β以饱和、剂量依赖的方式刺激IL-6和IL-11的分泌。可溶性IL-6受体的分泌与IL-6的分泌在组成性或存在IL-1β时均无相关性。在14个样本中的4个样本中，IL-1β也刺激了粒细胞-巨噬细胞集落刺激因子的分泌。在为期2周的培养期内，9个培养物中有7个未检测到IL-1β。在7个测试培养物中，IL-6未刺激IL-1β的分泌。细胞的存活和生长依赖血清，血清替代品（1%胰岛素-转铁蛋白-硒-牛血清白蛋白）无法维持其生长。在含有10%胎牛血清并添加1%胰岛素-转铁蛋白-硒-牛血清白蛋白的培养基中生长细胞分泌的IL-6比仅在血清中生长的细胞多10倍。7名接受雌激素替代疗法女性的骨髓中，IL-6（75%；P < 0.006）和IL-11（43%；P < 0.05）的组成性分泌低于年龄匹配的对照组骨髓，且外源性IL-1β对IL-6和IL-11分泌的刺激作用减弱。这些数据表明，人骨髓基质培养物产生细胞因子的模式因年龄和雌激素状态而异。
