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Objective:To investigate the effect of Yougui Yin on steroid-induced femoral head necrosis in rats and the regulation of Wnt/β-catenin signaling pathway, and to explore its mechanism.Methods:Fifty SD rats were randomly divided into normal group, model group, high-dose group (26.4 g/kg), medium-dose group (13.2 g/kg) and low-dose group (6.6 g/kg) of Yougui Yin, with 10 rats in each group. Except the normal group, the other groups were prepared with SANFH model by combining LPS and methylprednisolone injection. The treatment groups were intragastrically administered with Yougui Yin, once a day for 8 weeks. After the final administration, the serum calcium and phosphorus levels of rats in each group were determined by automatic biochemical analyzer. The femoral head specimens of rats in each group were detected by MRI, and the pathological changes of the femoral head were observed by HE staining. The expressions of caspase-3, β-catenin and Wnt3α mRNA in the femoral head of rats were detected by RT-PCR.Results:Compared with the model group, the levels of serum calcium and phosphorus in the medium and high dose groups were increased significantly ( P<0.05, P<0.01), and the level of serum phosphorus in the low dose group was increased significantly ( P<0.05); In the medium and high dose groups, the femoral empty bone lacuna rate was decreased ( P<0.05, P<0.01); The expression of caspase-3 mRNA (2.146±0.191, 1.688±0.247, 1.370±0.252 vs. 2.535±0.236) in the low, medium and high dose groups were decreased ( P<0.05, P<0.01), and the expression of β-catenin mRNA (0.433±0.102, 0.496±0.091, 0.698±0.089 vs. 0.259±0.106) were increased ( P<0.05, P<0.01); The expression of Wnt3α mRNA (0.509±0.061, 0.833±0.053 vs. 0.384±0.052) in the medium and high dose groups were increased ( P<0.05, P<0.01); In the medium and high dose groups, MRI showed higher T2 signals around the joint, and high T2 signals within the joint, which were clearly distinguished from the femoral head cartilage, and there was no obvious abnormal signal within the femoral head. HE staining in the Yougui Yin medium and high dose groups showed that trabecular bone was coarse and arranged regularly, most of the bone cells were normal, and empty bone lacunae were less. Conclusion:The protective effect of Yougui Yin on SANFH rats may be related to the inhibition of Wnt/β-catenin signaling pathway.
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BACKGROUND:Bone morphogenetic protein-7 can promote proliferation of bone marrow mesenchymal stem cels and has important significance in terms of inducing new bone formation. OBJECTIVE:To observe the protein and mRNA expression of bone morphogenetic protein-7 in bone marrow mesenchymal stem cels transfected with bone morphogenetic protein-7. METHODS:Rat bone marrow mesenchymal stem cels were isolated and cultured, and then identified by the morphology and cel surface markers. Lentiviral vectors carrying bone morphogenetic protein-7 were constructed to transfect bone marrow mesenchymal stem cels. After total RNA extraction, the protein and mRNA expression of bone morphogenetic protein-7 was detected by RT-PCR and western blot methods. RESULTS AND CONCLUSION:PLV-sfGFP(2A)-BMP7 recombinant plasmids were successfuly constructed and transferred into rat bone marrow mesenchymal stem cels. The mRNA and protein expression of bone morphogenetic protein-7 in the transfection group was significantly higher than that in the blank vector and blank groups (P < 0.05), indicating exogenous bone morphogenetic protein-7 gene is effectively integrated into the bone marrow mesenchymal stem cels.
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OBJECTIVE:To establish an HPLC method for determination of gentiopicroside in Gentian liver-purging pill. METHODS:The chromatographic separation was carried out on VP-ODS(150 mm?4.6 mm,5 ?m). The mobile phase consisted of acetonitrile-0.1% phosphoric acid solution(15∶85) at a flow rate of 1.0 mL?min-1.The UV detection wavele-ngth was set at 270 nm and column temperature was kept at 25 ℃.RESULTS:The liner range of gentiopicroside was 0.355 4～5.331 ?g(r=0.999 8) and the average recovery was 100.05%(RSD=2.2%,n=6). CONCLUSION:The method is simple,rapid and accurate,and it is applicable for the quality control of Gentian liver-purging pill.
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OBJECTIVE:To study the long-term toxicity of Reketing granules.METHODS:The experiment was performed in 4 groups:one control group(normal saline)and three Reketing subgroups(188 g?kg-1,94 g?kg-1,18 g?kg-1).After intragastrical administration for 3 consecutive months,the blood routine,urine routine and serum biochemical indicators were measured in rats,and the autopsy of the major organs was performed.After drug withdrawal for 2 weeks,the above indexes were measured again to find out the abnormality.RESULTS:After 3-month medication,the rats experienced a gradual increase in body weight and developed well.In Reketing high dose-treated group,glutamate-pyruvate transaminase(GPT)and blood urea nitrogen(BUN)levels increased slightly;and except for lesion of liver and kidney in small number of rats,the peripheral hemogram,urine routine and the blood biochemical indicators were all normal.The above indexes and the pathological findings of the organs after drug withdrawal for 2 weeks were all normal.CONCLUSION:Reketing granules can lead to mild lesion of liver and kidney of rats during medication,but the symptoms disappear after drug withdrawal.