目的 探讨兔骨髓间充质干细胞(MSCs)体外定向诱导分化为成骨细胞所需的地塞米松的最佳浓度.方法 抽取兔骨髓,采用全骨髓贴壁培养法体外培养,贴壁细胞进行传代.取第3代细胞,随机分为5组,在培养基中添加成骨分化诱导剂β-甘油磷酸钠、L-抗坏血酸,5组添加的地塞米松的浓度分别为1×10-6mol/L、1×10-7mol/L、1×10-8mol/L、1×10-9mol/L、1×10-10mol/L.培养10d后,在倒置显微镜观察细胞形态,计算细胞浓度,采用5-溴-4-氯-3-吲哚磷酸盐(BCIP)/氯化硝基四氮唑蓝(NBT)底物显色试剂检测碱性磷酸酶(ALP),茜素红染色检测钙结节.结果 较高浓度(1×10-8mol/L、1×10-7mol/L、1×10-6mol/L)的地塞米松相对于较低浓度(1×10-9mol/L、1×10-10mol/L)的地塞米松可显著增加ALP表达(P均<0.05),并可增加钙结节的形成;1×10-9mol/L及其以下浓度的地塞米松对细胞增殖无明显影响(P均>0.05),1×10-7mol/L以上浓度的地塞米松可明显抑制细胞的增殖(P<0.01).结论 地塞米松在体外定向诱导分化兔骨髓间充质干细胞为成骨细胞,最佳的浓度为1×10-8mol/L.%Objective It is to approach the establishment of the animal model of diabetes mellitus and observe the curative effect of traditional Chinese medicine Shutangbao. Methods All mice were divided into four groups: normal group, model group, treatment group 1 and treatment group 2. In last 3 groups, the diabetic mouse model was established with intraperitone-al injection of Alloxan according to 200 mg/kg body weight dose once, after 3 days apart, with intraperitoneal injection of STZ according to 40 mg/kg body weight for 4 days, once a day. When the FPG was increased the mice in treatment group 1 and treatment group 2 accepted intragastric administrations with Shutangbao and dianoguanil respectively. The blood glucose levels were detected before making model, after establishment of model, after treatment. The end of the experiment, blood was collected from the eye to detect the content of total cholesterol ( TC ) and triglyceride ( TG ). Results In normal group, model group, treatment group 1 and treatment group 2, the blood glucose levels was ( 6. 25 ± 1.21), ( 13.72 ±3. 87 ), ( 18. 54 ± 9. 83 ), ( 15. 86 ± 6. 17 ) mmol/L respectively after modeling for 2 days, there were significant differences between the last 3 groups and normal group (P<0. 05 or 0.01 ); and was ( 5. 74 ± 1. 19 ), ( 17.03 ±10.22), ( 19. 67 ± 4. 97 ), ( 17.09 ± 5. 51 ) mmol/L respectively after modeling for 5 days, there were very significant differences between them ( all P <0. 01 ); and was ( 6. 99 ± 0. 92 ), ( 17.14 ± 8. 69 ), ( 16. 79 ± 7. 00 ), ( 10. 99 ± 2. 24 ) mmol/L respectively after treatment for 24 days, the level in treatment group 2 was reduced obviously, and had a certain degree to reduce in treatment group 1 also. There was no significant difference in levels of TC and TG among all groups. Conclusion The curative effect of Shutangbao for diabetes mellitus is better than that of dianoguanil.
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