Objective To explore effect of platelet-rich plasma (PRP) on rabbit BMSCs differentiation into SC in vitro and to detect secretory function of the differentiated cells. Methods BMSCs isolated from 5 mL bone marrow of 2-montholdNew Zealand white rabbit were cultured using density gradient centrifugation and adherence screening methods. A total of 5 mL femoral vein blood was obtained from rabbits to prepare PRP using modified Appel method. The BMSCs at passage 3 were divided into three groups: the combined induction group, in which the cells were cultured with complete medium containing PRP after β-mercaptoethanol and retinoic acid inductions; the simple induction group, in which the cells were cultured with L-DMEM complete medium without PRP afterβ-mercaptoethanol and retinoic acid induction; the control group, in which the cells were cultured with L-DMEM complete medium. Growth condition of the cells in each group was observed using inverted microscope. cell identification was conducted at 4, 7, 9, and 11 days after culture using immunofluorescence staining method, and NGF content was detected by ELISA method. NGF mRNA expression was assayed by RT-PCR 11 days after culture. Results Most cells in the combined induction and the simple induction group were out of BMSCs typical cell morphology 4 days after culture; cells in the combined induction group were out of BMSCs typical cell morphology and changed into cells resembl ing SC in terms of morphology and contour 9 days after culture. The cells in the control group showed no obvious morphological changes. S-100 protein expression in the cells was evident in the combined induction and the simple induction group at each time point after induced culture; the positive expression rate of cell in each group was increased over time, and significant differences were evident between the combined induction group and the simple induction group 7, 9, and 11 days after culture (P lt; 0.05). Control groupwas negative for the expression. There were significant differences when comparing the control group with the combined induction group or the simple induction group in terms of NGF content at each time point (P lt; 0.01). Significant difference was evident between the combined induction group and the simple induction group 7, 9, and 11 days after culture (P lt; 0.05), and no significant difference was noted 4 days after culture (P gt; 0.05). Relative intensity of NGF mRNA expression in the combined induction group was greater than that of the simple induction group 11 days after culture (P lt; 0.05). Conclusion Rabbit BMSCs can differentiate into SC excreting NGF under certain induction condition in vitro. PRP can remarkably promote BMSCs differentiation into SC.
目的 观察罗格列酮钠对血糖控制未达标的2型糖尿病(T2DM)合并与不合并非酒精性脂肪肝(NAFL)患者的降糖疗效和安全性。 方法 2009年1月-2011年1月60例仅用磺脲类和二甲双胍治疗血糖控制未达标的T2DM患者,按合并和不合并NAFL分为观察组和对照组各30例,两组均在原口服降糖药基础上联合加用国产罗格列酮钠4 mg 1次/d,治疗共3个月,观察治疗前后的血糖、胰岛素、糖化血红蛋白(HbA1c)、体质量指数(BMI)、血脂、肝功、血压水平以及药物不良反应,并比较治疗后的血糖达标率。 结果 两组患者治疗后的空腹血糖(FPG)、餐后2 h血糖(2hPG)、HbA1c、空腹胰岛素、甘油三酯和极低密度脂蛋白胆固醇均较治疗前下降,高密度脂蛋白胆固醇较治疗前升高(P<0.05),而丙氨酸转氨酶、总胆固醇、低密度脂蛋白胆固醇及血压无明显变化(P>0.05),但观察组治疗后的FPG和2hPG均较对照组下降更明显(P<0.01),且血糖达标率为73.3%,显著高于对照组的46.7%(P<0.05),同时观察组餐后2 h胰岛素(2hINS)水平在治疗前后均明显高于对照组而且治疗后有显著下降(P<0.01),但对照组治疗后2hINS虽然也有下降但无统计学意义(P>0.05)。观察组治疗前后BMI无明显变化,但对照组治疗后BMI有明显的升高(P<0.05)。结论 国产罗格列酮钠片对血糖控制未达标的T2DM合并和不合并NAFL患者均有进一步降低血糖、HbA1c以及改善血脂的作用,但对T2DM合并NAFL的患者的降糖疗效更显著,未见加重肝功能损坏,不良反应小,可作为此类患者联合用药的一种选择。