Li H, Miyahara T, Tezuka Y, Namba T, Suzuki T, Dowaki R, Watanabe M, Nemoto N, Tonami S, Seto H, Kadota S
Institute of Natural Medicine, Toyama Medical and Pharmaceutical University, Japan.
Biol Pharm Bull. 1999 Apr;22(4):391-6. doi: 10.1248/bpb.22.391.
We previously isolated berberine from aqueous extracts of tsu-kan-gan, a Kampo formula used for the treatment of osteoporosis. Berberine caused an inhibitory effect on parathyroid hormone (PTH)-stimulated bone resorption in neonatal mouse bone. In this report we describe the inhibitory effect of berberine on the formation of osteoclast-like multinucleated cells (OCLs) in the co-culture of mouse osteoblastic cells and bone marrow cells in the presence of 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3], PTH and interleukin-1alpha (IL-1alpha). Berberine dose-dependently inhibited the formation of tartrate-resistant acid phosphatase (TRAP)-positive OCLs induced by 1alpha25(OH)2D3, PTH and IL-1alpha. We prepared OCLs in the co-culture of osteoblastic cells and bone marrow cells. The effect of berberine on pit formation by OCLs was examined using dentin slices. As OCLs are terminally differentiated multinucleated cells, the survival of OCLs affects the bone-resorbing activity of OCLs. This prompted us to count the number of TRAP-positive OCLs on the slices. Berberine dose-dependently inhibited pit formation and caused a decrease in the number of TRAP-positive OCLs. Calcitonin (CT) inhibited pit formation without affecting the number of OCLs. Berberine accelerated the cell death in OCLs cultivated on a culture plate, but CT did not affect the cell death of OCLs. This suggests that the decrease in the number of OCLs on dentin slices may be due to apoptotic cell death in OCLs. In fact, Hoechst 33258 staining revealed that the treatment of OCLs with berberine resulted in condensed nuclei and a decrease in cell size. Oral administration of the berberine (30 and 50 mg/kg/d) to ovariectomized rats prevented a decrease in bone mineral density (BMD) of the lumbar vertebra without affecting the weight of the uterus and plasma concentration of estradiol. These results suggested that berberine prevented a decrease in BMD in vivo by inhibiting osteoclastic bone resorption.
我们之前从用于治疗骨质疏松症的汉方药“津肝丸”的水提取物中分离出了黄连素。黄连素对新生小鼠骨骼中甲状旁腺激素（PTH）刺激的骨吸收具有抑制作用。在本报告中，我们描述了黄连素在1α,25 - 二羟基维生素D3 [1α,25(OH)2D3]、PTH和白细胞介素 - 1α（IL - 1α）存在的情况下，对小鼠成骨细胞和骨髓细胞共培养体系中破骨细胞样多核细胞（OCLs）形成的抑制作用。黄连素剂量依赖性地抑制了由1α,25(OH)2D3、PTH和IL - 1α诱导的抗酒石酸酸性磷酸酶（TRAP）阳性OCLs的形成。我们在成骨细胞和骨髓细胞的共培养体系中制备了OCLs。使用牙本质切片研究了黄连素对OCLs形成陷窝的影响。由于OCLs是终末分化的多核细胞，OCLs的存活会影响其骨吸收活性。这促使我们对切片上TRAP阳性OCLs的数量进行计数。黄连素剂量依赖性地抑制陷窝形成，并导致TRAP阳性OCLs数量减少。降钙素（CT）抑制陷窝形成，但不影响OCLs的数量。黄连素加速了培养板上培养的OCLs的细胞死亡，但CT不影响OCLs的细胞死亡。这表明牙本质切片上OCLs数量的减少可能是由于OCLs中的凋亡性细胞死亡。事实上，Hoechst 33258染色显示，用黄连素处理OCLs会导致细胞核浓缩和细胞大小减小。对去卵巢大鼠口服黄连素（30和50 mg/kg/d）可防止腰椎骨矿物质密度（BMD）降低，而不影响子宫重量和雌二醇的血浆浓度。这些结果表明，黄连素通过抑制破骨细胞骨吸收，在体内防止了BMD的降低。
