Yamaguchi M, Kishi S
Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, Japan.
Peptides. 1995;16(8):1483-8. doi: 10.1016/0196-9781(95)02030-6.
The effect of transforming growth factor-beta (TGF-beta) on osteoclast-like cell formation in mouse marrow culture in vitro was investigated. The bone marrow cells were cultured for 7 days in alpha-minimal essential medium containing a well-known bone resorbing agent. Osteoclast-like cell formation was estimated with staining for tartrate-resistant acid phosphatase (TRACP), a marker enzyme of osteoclasts. The presence of TGF-beta (10(-13)-10(-11) M) caused a significant increase in the number of osteoclast-like multinucleated cells (MNCs); the maximum effect was seen with 10(-12) MTGF-beta. With a higher concentration (10(-10) M) of TGF-beta, the growth factor dramatically inhibited the 1,25-dihydroxyvitamin D5 [1,25(OH)2D3; 10(-8) M]-induced formation of osteoclast-like MNCs. This inhibitory effect was also seen in the formation of osteoclast-like MNCs stimulated by parathyroid hormone (10(-8) M), prostaglandine E2 (10(-6) M), and interleukin-1 alpha (50 U/ml). The stimulatory effect of TGF-beta (10(-12) M) on osteoclast-like MNCs formation was inhibited by zinc sulfate (10(-6) M) or zinc-chelating dipeptide [beta-alanyl-L-histidinato zinc (AHZ), 10(-6) M]. The stimulating effect of TGF-beta was markedly weakened by the presence of EGTA (0.5 mM), a chelator of Ca2+. The inhibitory effect of zinc compounds was not seen in the presence of EGTA. Moreover, the inhibitory effect of TGF-beta (10(-10) M), zinc sulfate (10(-6) M), or AHZ (10(-6) M) on osteoclast-like MNCs formation was not demonstrated in mature osteoclastic cells, although calcitonin (3 x 10(-8) M) significantly inhibited the osteoclastic formation. The present study demonstrates that TGF-beta has a stimulating and an inhibiting effect on osteoclast-like cell formation in mouse marrow culture, and that zinc can inhibit the stimulatory effect of TGF-beta.
研究了转化生长因子-β（TGF-β）对体外培养的小鼠骨髓中破骨细胞样细胞形成的影响。将骨髓细胞在含有一种知名骨吸收剂的α-最低必需培养基中培养7天。用抗酒石酸酸性磷酸酶（TRACP）染色来评估破骨细胞样细胞的形成，TRACP是破骨细胞的一种标记酶。TGF-β（10^(-13)-10^(-11) M）的存在导致破骨细胞样多核细胞（MNCs）数量显著增加；在10^(-12) M TGF-β时观察到最大效应。当TGF-β浓度较高（10^(-10) M）时，该生长因子显著抑制1,25-二羟基维生素D5 [1,25(OH)2D3；10^(-8) M]诱导的破骨细胞样MNCs形成。在甲状旁腺激素（10^(-8) M）、前列腺素E2（10^(-6) M）和白细胞介素-1α（50 U/ml）刺激形成破骨细胞样MNCs的过程中也观察到了这种抑制作用。TGF-β（10^(-12) M）对破骨细胞样MNCs形成的刺激作用被硫酸锌（10^(-6) M）或锌螯合二肽[β-丙氨酰-L-组氨酸锌（AHZ），10^(-6) M]抑制。EGTA（0.5 mM），一种Ca2+螯合剂的存在显著削弱了TGF-β的刺激作用。在EGTA存在的情况下未观察到锌化合物的抑制作用。此外，尽管降钙素（3×10^(-8) M）显著抑制破骨细胞形成，但TGF-β（10^(-10) M）、硫酸锌（10^(-6) M）或AHZ（10^(-6) M）对成熟破骨细胞中破骨细胞样MNCs形成的抑制作用未得到证实。本研究表明，TGF-β对小鼠骨髓培养中破骨细胞样细胞的形成具有刺激和抑制作用，并且锌可以抑制TGF-β的刺激作用。
