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目的:研究体外左、右归丸对去卵巢大鼠骨髓间充质干细胞经地塞米松诱导后凋亡的影响。方法:将60只SPF级2月龄SD雌性未生育大鼠随机分为6组,分别是空白对照组(CON)、模型组(OVX)、假手术组(SHAM)、左归丸组(ZGP)、右归丸组(YGP)和补佳乐组(BJL)组。采用双侧去卵巢法建立绝经后骨质疏松症模型,术后3 d灌胃饲养,按每千克体重大鼠的给药量为人的6.3倍计算,每日灌胃1次,给药12周后,利用全骨髓贴壁法获取去卵巢大鼠BMSCs,进行体外培养。对去卵巢大鼠BMSCs进行干预,采用流式细胞仪通过表面抗原CD29、CD90、CD11b/c鉴定BMSCs,模型组分别加入10~(-6)mol/L、10~(-5)mol/L、5×10~(-5)mol/L地塞米松,分别作用24、48、72 h后通过流式细胞仪检测凋亡率。并用流式细胞仪检测比较以上6组BMSCs以10~(-6)mol/L地塞米松诱导48 h后的凋亡率。结果:流式鉴定CD29、CD90均显示为阳性,CD11b/c为阴性;10~(-6)mol/L地塞米松作用48 h时与其他组比较,去卵巢大鼠BMSCs的凋亡率明显升高(P<0.05),并以此时间、浓度作为诱导条件。与OVX组相比,SHAM组、ZGP组、YGP组和BJL组凋亡率降低,有显著性差异(P<0.05);与ZGW组比较,YGW组凋亡率较高(P<0.05)。结论:左、右归丸均能抑制去卵巢大鼠BMSCs的凋亡,且左归丸的效果优于右归丸。
OBJECTIVE: To study the effect of Zuogui Yougui on the apoptosis of bone marrow mesenchymal stem cells induced by dexamethasone in ovariectomized rats. Methods: Sixty SPF female SD rats of 2 months old were randomly divided into 6 groups: control group (CON), model group (OVX), sham operation group (SHAM), Zuoguiwan group ), Yougui pill group (YGP) and nociception group (BJL) group. Bilateral ovariectomized osteoporosis model was established by postmenopausal ovariectomized rats. The rats were fed intragastrically 3 days after operation. The dosage was 6.3 times as much as that of human. The rats were given gavage once daily for 12 weeks BMSCs from ovariectomized rats were obtained by whole bone marrow adherent method and cultured in vitro. BMSCs were isolated from ovariectomized rats. BMSCs were identified by surface antigen CD29, CD90 and CD11b / c using flow cytometry. The model group was treated with 10 -6 mol / L 10 -5 mol / L , 5 × 10 ~ (-5) mol / L dexamethasone. After 24, 48 and 72 h, the apoptotic rate was detected by flow cytometry. Flow cytometry was used to detect the apoptosis rate of the above 6 groups of BMSCs induced by 10 -6 mol / L dexamethasone for 48 h. Results: Flow cytometry showed CD29 and CD90 were positive and CD11b / c was negative. Compared with other groups, apoptosis rate of ovariectomized BMSCs was significantly higher at 10 ~ (-6) mol / L dexamethasone for 48 h (P <0.05), and the time and concentration as the induction conditions. Compared with OVX group, the apoptotic rates in SHAM group, ZGP group, YGP group and BJL group were significantly decreased (P <0.05). Compared with ZGW group, the apoptosis rate in YGW group was higher (P <0.05). Conclusion: Both left and right Gui Wan can inhibit the apoptosis of BMSCs in ovariectomized rats, and Zuo Gui Wan is better than You Gui Wan.