[Effect of Hydroxy Safflower Yellow A on glucocorticoid-induced bone marrow mesenchymal stem cells osteogenic differentiation].

OBJECTIVE: To observe the effect of Hydroxy Safflower Yellow A (HSYA) on the expression of osteogenic markers, such as alkaline phosphatase, Cbf(alpha)l and type I collagen, and explore the mechanism of HSYA in the prevention and treatment of glucocorticoid-induced ischemic necrosis of femoral head. METHODS: Fifteen healthy and adult New Zealand white rabbits were collected and weighted 0.9 to 1.3 kg. The rabbits were injected abdominally with anesthetic drugs, then received marrow cavity puncture of tibia and anterior superior iliac spine to get bone marrow blood. Rabbits bone marrow mesenchymal stem cells (BMSCs) were separated from the bone marrow blood, cultured in vitro and passaged. The 3rd generation of BMSCs which had good growth condition were randomly divided into blank group, model group and HSYA groups with different doses. The BMSCs in model group were treated with high dose of dexamethasone to induce adipogenic differentiation of cells cultured in vitro, and inhibit osteogenic differentiation. The BMSCs in HSYA groups received high dose of dexamethasone and different concentrations of HSYA simultaneously. The blank group received not any special handling. After a week,the expressions of alkaline phosphatase, Cbf(alpha)l and type I collagen mRNA were detected. RESULTS: The alkaline phosphatase activity was significantly decreased in BMSCs of the model group as compared with the blank group (P < 0.01), and the expression of Cbf(alpha)l and type I collagen mRNA were also decreased significantly (P<0.01). The alkaline phosphatase activity was significantly increased in BMSCs of each HSYA group as compared with the model group (P < 0.05 or P < 0.01), and the expression of Cbf(alpha)l and type I collagen mRNA were also increased significantly (P < 0.05 or P < 0.01). CONCLUSION: The mechanism of HSYA may be related to the effect of antagonism to the reduced osteogenic differentiation induced by glucocorticoid.

[Influence of ground beetle on steroid-induced BMSCs osteogenic differentiation].

OBJECTIVE: To investigate the pathogenesis of steroid-induced avascular necrosis of femoral head and the control mechanisms of Ground Beetle. METHODS: The bone marrow-derived mesenchymal stem cells were cultured in vitro. The BMSCs in the 3rd generation were randomly divided into blank group, model group and Chinese medicine low, medium and high dose group. BMSCs of model group using high-dose steroid-induced in vitro were adipogenic differentiation to inhibit osteogenesis. Chinese medicine low, medium and high-dose groups at the same time were given the Ground Beetle intervention serum containing insects. The expression of osteocalcin, Alkaline phosphatase and Type I Collagen mRNA were detected and interfered 6 days later. RESULTS: Serum containing Ground Beetle could reverse content of steroid-induced alkaline phosphatase of BMSCs, the expression of ostercalcin and Type I Collagen mRNA decreased. CONCLUSION: The control mechanism of Ground Beetle on steroid-induced avascular necrosis is not only to improve the microcirculation, but also to inhibit steroid-induced osteogenic differentiation of BMSCs reduced.

[Effect on transforming growth factor-beta1 of glucocorticoid-induced avascular necrosis of femoral head in rats by treatment of activating blood circulation of Chinese herbal medicine].

OBJECTIVE: To establish rat models of Steroid-avascular necrosis of femoral head, observe the effects of activating blood circulation of chinese herbal medicine on genetic expression of transforming growth factor-beta1 (TGF-beta1). To interpret the mechanism of the effect on Steroid-avascular necrosis of femoral head by activating blood circulation,and offer a effective method to clinical. METHODS: Cleaner-40 SD rats,half males and half females, weight (200 +/- 20) g, were randomly divided into 2 groups: 4 rats were in common group and 36 rats were in medel group. The rats in medel group were administered with 24.5 mg/kg hydroxyprednisone twice a week peritoneal injection for 6 weeks induced to femur head necrosis. The rats in common group were through gluteus injection as control. There were 4 rats were killed in each group after 6 weeks, to be assure that the model were succed. All surplus rats were divided into treatment group and control group:the treatment group were administrated with activating blood circulation of Chinese herbal medicine 12.3 ml/kg per day,the control group were administrated with sodium chloride 12.3 ml/kg per day. Then, after 6 and 8 weeks, killed the animal and detected all indexes. RESULTS: (1) The expression of TGF-beta by immunohistochemistry and image analysis: the expression of femoral head TGF-beta increased significantly in treatment group than in control group and two group had significant differences. (2) Serum levels of TGF-beta1: in the treatment group serum expression of TGF-beta1 increased,compared with the control group had significant difference (P < 0.01). (3) The femoral head local TGF-beta1 mRNA transcription: treatment group in the first six weeks, expressed that the increase in the first eight weeks, expressed also reduced,and the control group in the first six weeks, expressed a decrease in the first eight weeks,was not detected to TGF-beta1 mRNA expression of difference between the two groups was significant (P < 0.01). TGF-beta1 mRNA in serum and the femoral head with local expression of TGF-beta1, in consistency. CONCLUSION: Rat abdominal cavity prednisolone acetate injection plus intermittent standing avascular necrosis modeling stability, good repeatability. Taohongsiwu through the promotion of hormone-ischemic necrosis of the femoral head rat model of TGF-beta1 mRNA transcription, and promote expression of TGF-beta1.

Effect of Chinese drugs for promoting blood circulation and eliminating blood stasis on vascular endothelial growth factor expression in rabbits with glucocorticoid-induced ischemic necrosis of femoral head.

OBJECTIVE: To probe into the mechanism of Chinese drugs for promoting blood circulation and eliminating blood stasis in the prevention and treatment of glucocorticoid-induced ischemic necrosis of femoral head. METHODS: Thirty New Zealand adult white rabbits were randomly divided into a normal control group (n=5) and a model group (n=25). Hydroxyprednisone acetate was intramuscularly administered to the rabbits in the model group in a dosage of 7.5 mg/kg, twice per week for 6 weeks, to induce ischemic necrosis of femoral head and normal saline of the equal volume was intramuscularly administered to the rabbits in the normal control group, twice per week for 6 weeks. Then, the 5 rabbits from the normal control group and 5 rabbits selected randomly from the model group were sacrificed and the changes in histopathology and the expression of Vascular Endothelial Growth Factor (VEGF) were observed. The other 20 rabbits in the model group were randomly divided into the treatment group 1 and the treatment group 2, and the control group 1 and the control group 2, 5 rabbits in every group. Taohong Siwu Tang ([Chinese characters: see text] Decoction of Four Drugs with Addition of Peach Kernel and Safflower) was orally administered to the rabbits in the treatment group 1 and the treatment group 2 in a dosage of 7 ml/kg, once daily and normal saline of the equal volume was orally administered to the rabbits in the control group1 and the control group, 2 once daily. After 10 weeks the rabbits in the treatment group 1 and the control group1 were sacrificed and after 13 weeks the rabbits in the treatment group 2 and the control group 2 were sacrificed, and the expression of VEGF was detected in these rabbits. RESULTS: The expression of VEGF was significantly enhanced in rabbits of the model group as compared with the normal control group (P < 0.01), and gradually reduced with the lapse of time. The expression of VEGF in the control groups was significantly reduced as compared with the treatment groups (P < 0.001). CONCLUSION: Chinese drugs for promoting blood circulation and eliminating blood stasis can improve the microcirculation of femoral head in rabbits with glucocorticoid-induced ischemic necrosis of femoral head by promoting the expression of VEGF.