Seropharmacological study on osteogenic effects of post-absorption ingredients of an osteoprotective herbal formula.
OBJECTIVE
To further investigate the {ptin vitro} effects of an osteoprotective herbal formula "ELP" (Herba Epimedii, Fructus Ligustri Lucidi and Fructus Psoraleae) using seropharmacological approach.


METHODS
Rats were fed with ELP or its individual component herbs for 2 days. The serum containing the postabsorbed ingredients of the herbal items were collected for cell culture using UMR106 cell, RAW264.7 cell and mesenchymal stem cell (MSC) isolated from the bone marrow of the rats. The effects of the herbal-containing serum on cell toxicity were detected by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay; bromodeoxyuridine assay was conducted to measure the cell proliferation of UMR106 cell and MSC; cell activity was measured using colorimetric method, and mRNA expression of runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and osteopontin (OPN) of UMR106 and MSC as well as matrix metalloproteinase 9 (MMP-9), tartrate-resistant acid phosphatase (TRAP) and cathepsin K of RAW264.7 were analyzed using real-time reverse-transcription polymerase chain reaction.


RESULTS
ELP and its component serum exhibited no cytotoxic effects on the cells. The ELP-containing serum increased the proliferation of UMR106 cell and MSC by 25.7% and 14.4 %, respectively and the alkaline phosphatase activity of MSC was increased by 42.6%. On the contrary, it inhibited the RAW264.7 cell differentiation by 29.2 %. ELP serum upregulated the Runx2 expression of UMR and MSC by 1.18 fold and 1.27 fold, respectively. It also upregulated ALP and OPN expression in MSC by 1.69- and 2.12-fold, respectively. On the other hand, ELP serum down-regulated MMP-9 and cathepsin K expression of RAW264.7 cell by 0.46- and 0.36-fold, respectively.


CONCLUSIONS
The serum of the animals fed with ELP contains active ingredients which are effective in promoting osteogenesis and inhibiting osteoclastogenesis.
Institute of Chinese Medicine, Chinese University of Hong Kong, Hong Kong, China.|State Key Laboratory of Phytochemistry and Plant Resources in West China, Chinese University of Hong Kong, Hong Kong, China.|Department of Orthopaedics and Traumatology, Chinese University of Hong Kong, Hong Kong, China.
Wing-Sum
WS
Siu
Institute of Chinese Medicine, Chinese University of Hong Kong, Hong Kong, China.|State Key Laboratory of Phytochemistry and Plant Resources in West China, Chinese University of Hong Kong, Hong Kong, China.
Chun-Hay
CH
Ko
Institute of Chinese Medicine, Chinese University of Hong Kong, Hong Kong, China.|State Key Laboratory of Phytochemistry and Plant Resources in West China, Chinese University of Hong Kong, Hong Kong, China.
Hing-Lok
HL
Wong
Institute of Chinese Medicine, Chinese University of Hong Kong, Hong Kong, China.|State Key Laboratory of Phytochemistry and Plant Resources in West China, Chinese University of Hong Kong, Hong Kong, China.
Si
S
Gao
Institute of Chinese Medicine, Chinese University of Hong Kong, Hong Kong, China.|State Key Laboratory of Phytochemistry and Plant Resources in West China, Chinese University of Hong Kong, Hong Kong, China.
Wai-Ting
WT
Shum
Institute of Chinese Medicine, Chinese University of Hong Kong, Hong Kong, China.|State Key Laboratory of Phytochemistry and Plant Resources in West China, Chinese University of Hong Kong, Hong Kong, China.
Clara Bik-San
CB
Lau
Department of Orthopaedics and Traumatology, Chinese University of Hong Kong, Hong Kong, China.
Lung-Kim
LK
Hung
Institute of Chinese Medicine, Chinese University of Hong Kong, Hong Kong, China. pingcleung@cuhk.edu.hk.|State Key Laboratory of Phytochemistry and Plant Resources in West China, Chinese University of Hong Kong, Hong Kong, China. pingcleung@cuhk.edu.hk.|Department of Orthopaedics and Traumatology, Chinese University of Hong Kong, Hong Kong, China. pingcleung@cuhk.edu.hk.
Ping-Chung
PC
Leung
D065567:Absorption, Physiological / Q000187:drug effects*; D000818:Animals; D001842:Bone and Bones / Q000187:drug effects* / Q000473:pathology; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D008297:Male; D051379:Mice; D010010:Osteoclasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D010012:Osteogenesis / Q000187:drug effects*; D020011:Protective Agents / Q000494:pharmacology*; D000067996:RAW 264.7 Cells; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D044967:Serum / Q000378:metabolism*
D004365:Drugs, Chinese Herbal; D020011:Protective Agents; D012333:RNA, Messenger
herbal formula; osteogenesis; osteoporosis; seropharmacology