The Effect of Kampo Formulae on Bone Resorption in Vitro and in Vivo. II. Detailed Study of Berberine

• Published in: Biological & pharmaceutical bulletin 1999/04/15, Vol.22(4), pp.391-396
• Main Authors: LI, Huiying, MIYAHARA, Tatsuro, TEZUKA, Yasuhiro, NAMBA, Tsuneo, SUZUKI, Takayuki, DOWAKI, Risa, WATANABE, Makoto, NEMOTO, Nobuo, TONAMI, Syuichi, SETO, Hikaru, KADOTA, Shigetoshi
• Format: Article
• Language: English
• Subjects: Animals; Anti-Bacterial Agents - pharmacology; antiosteoporotic; Apoptosis - drug effects; berberine; Berberine - pharmacology; Biological and medical sciences; Body Weight - drug effects; Bone Density - drug effects; Bone Resorption; Bones, joints and connective tissue. Antiinflammatory agents; Calcitonin - pharmacology; Calcitriol - pharmacology; Drugs, Chinese Herbal - pharmacology; Female; General pharmacology; Interleukin-1 - pharmacology; Macrolides; Male; Medical sciences; Mice; Organ Size - drug effects; osteoclast; osteoclastogenesis; Osteoclasts - cytology; Osteoclasts - drug effects; Ovariectomy; ovariectomy (OVX); Parathyroid Hormone - pharmacology; Pharmacognosy. Homeopathy. Health food; Pharmacology. Drug treatments; pit formation; Rats; Rats, Wistar; Uterus - drug effects
• ISSN: 0918-6158; 1347-5215
• 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 1α, 25-dihydroxyvitamin D3 [1α, 25(OH)2D3], PTH and interleukin-1α (IL-1α). Berberine dosedependently inhibited the formation of tartrate-resistant acid phosphatase (TRAP)-positive OCLs induced by 1α, 25 (OH)2D3, PTH and IL-1α. We prepared OCLa 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 calls, 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. Calctinin (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 suggestes 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 ras prevented a decrease in bone mineral density (BMD) of the lumber 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.