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目的:研究左归丸、右归丸及其拆方对去卵巢大鼠骨髓间充质干细胞(bone marrow stem cells,BMSCs)成脂诱导的影响,并探讨其相关的作用机制。方法:将90只SD雌性大鼠,随机选取10只作为正常组,其余80只再随机选取10只经双侧腰背部切口切除卵巢周围少量脂肪组织作为假手术组,其余70只以相同切口切除双侧卵巢作为模型组,分别以正常组,假手术组,模型组,左归丸组(18.9 g·kg~(-1)),右归丸组(20.52 g·kg~(-1)),共同药组(15.12 g·kg~(-1)),滋阴药组(12.96 g·kg~(-1)),补阳药组(8.1 g·kg~(-1))和戊酸雌二醇片组(0.36 mg·kg~(-1))共9组,对BMSCs进行干预,流式细胞仪鉴定BMSCs,用油红O检测脂滴的形成;采用逆转录-聚合酶链式反应(RT-PCR)法检测过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptorγ,PPARγ),脂蛋白脂肪酶(lipoprotein,LPL),脂肪酸结合蛋白4(fatty acid binding protein 4,FABP4)mRNA的表达;采用蛋白质免疫印迹(Western blot)法检测CCAAT/增强子结合蛋白β(CCAAT/enhancer binding proteinβ,C/EBPβ)蛋白的表达。结果:与正常组比较,模型组BMSCs明显向脂肪细胞分化,PPARγ,LPL,FABP4 mRNA表达及C/EBPβ蛋白的表达明显升高(P<0.05);与模型组比较,左归丸、右归丸及其拆方均可抑制BMSCs向脂肪细胞分化,其中右归丸组可明显下调PPARγ,LPL,FABP4 mRNA,降低C/EBPβ的蛋白的表达(P<0.05)。结论:左归丸、右归丸及其拆方均能抑制BMSCs的成脂分化,且右归丸抑制成脂分化的作用优于左归丸,以及补阳药及二者的共同药组。
Objective: To study the effect of Zuogui Pill, Yougui Pill and its decomposed formulas on the adipogenic induction of bone marrow stem cells (BMSCs) in ovariectomized rats, and to explore its related mechanism. Methods: Totally 90 SD female rats were randomly selected as the normal group. The other 80 rabbits were randomly selected to receive a small amount of adipose tissue around the ovary for the removal of a small amount of adipose tissue. The other 70 rabbits were resected by the same incision The ovariectomized rats were divided into two groups: normal group, sham operation group, model group, Zuoguiwan group (18.9 g · kg -1), Youguiwan group (20.52 g · kg -1) , The common medicine group (15.12 g · kg -1), the yin yin medicine group (12.96 g · kg -1), the BuYangYaoGroup (8.1 g · kg -1) and the valerate BMSCs were treated with estradiol group (0.36 mg · kg -1) for 9 days. BMSCs were identified by flow cytometry. Lipid droplets were detected by oil red O staining. Reverse transcriptase-polymerase chain reaction The expressions of peroxisome proliferator-activated receptorγ (PPARγ), lipoprotein (LPL), fatty acid binding protein 4 FABP4) mRNA was detected by RT-PCR. The expression of CCAAT / enhancer binding proteinβ (C / EBPβ) protein was detected by Western blot. Results: Compared with the normal group, the BMSCs in the model group differentiated into adipocytes obviously, the expression of PPARγ, LPL and FABP4 mRNA and the expression of C / EBPβ protein were significantly increased (P <0.05). Compared with the model group, Pill and its disassembled prescriptions could inhibit the differentiation of BMSCs into adipocytes. Yougui pill group could obviously down-regulate the mRNA expression of PPARγ, LPL and FABP4 and decrease the protein expression of C / EBPβ (P <0.05). Conclusion: Zuoguiwan, Yougui Wan and its disassembled prescriptions can inhibit the adipogenic differentiation of BMSCs, and Yougui pill inhibit adipogenic differentiation better than Zuogui pill, as well as yang supplement pills and the two common drug group.