Objective: To observe the effects of Huangqi San formula (HQS) drug containing serum on osteoblasts (Ob) functions and related genes of diabetic osteoporosis under the condition of high glucose

and discuss the therapeutic mechanism of HQS on diabetic osteoporosis. Method: Ten 5-week-old healthy clean grade Wistar rats were randomly divided into the normal diet group and medicine group. The drug containing serum of HQS and control serum were prepared by serum pharmacology means

four neonatal healthy clean grade SD rats were used for primary culture of Obs. The cells were isolated by enzyme digestion and induced from skull bone of these SD neonatal rats. Cell counting kit (CCK-8) assay was used to detect the effect of drug containing serum (four different concentrations: 5%

10%

20% and 30%) on cell proliferation under high glucose condition (25.5 mmol·L-1). The serum concentration which was optimal for cells proliferation was then selected. Real-time PCR and Western blot assay were used to detect the effects of HQS drug containing serum on the expression levels of osteogenic genes osteopontin (OPN)

osteocalcin(OC)

alkaline phosphatase (ALP)

pancrease-bone related gene osteoprotegrin (OPG)

and Fork transcription factor 1 (FoxO1). Result: In cell proliferation experiments

10% and 30% serum concentration groups had significant higher results as compared with their blank serum groups (P<0.05)

and 20% serum concentration group also had significant higher results (P<0.01). In the Real-time PCR results

5% serum concentration group could significantly down-regulate OPN and OPG mRNA expression levels as compared with the blank control group (P<0.05). 10% serum concentration groups could significantly up-regulate OC and ALP mRNA expression levels (P<0.05)

and also significantly increase OPN mRNA expression (P<0.01)

but significantly down-regulate the OPG and FoxO1 mRNA expression levels as compared with the blank serum groups (P<0.01). Under 20% serum concentration

the differences were identical with those under 10% serum concentration

especially the differences in OPN

OPG and FoxO1 mRNA expression levels were more significant under 20% serum concentration. 20% serum concentration group could significantly up-regulate OC and OPN protein expression levels as compared with the blank control serum group (P<0.01)

while OPG protein expression was significantly down-regulated (P<0.01). Conclusion: The therapeutic mechanism of HQS may be related to its proliferation effects on osteoblasts under high glucose condition and its regulatory effects on the expression levels of OPG and FoxO1.