左归丸和右归丸对去卵巢大鼠股骨骨髓PPARγ、C/EBPβ、C/EBPα蛋白表达的影响
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摘要
目的探讨左归丸和右归丸治疗绝经后骨质疏松症的可能作用机制。方法 60只大鼠随机分为空白组、假手术组、模型组、补佳乐组、左归丸组、右归丸组,每组10只。除空白组、假手术组外,其余各组经双侧卵巢切除手术建立绝经后骨质疏松症大鼠模型。3天后,左归丸组、右归丸组、补佳乐组给予相应药物灌胃,剂量分别为9.45 g/(kg·d)、10.26 g/(kg·d)、0.09 g/(kg·d),空白组、假手术组、模型组灌胃2ml蒸馏水,各组均灌胃12周。灌胃结束后,HE染色观察股骨近端骨组织形态学变化,western blot法检测股骨骨髓过氧化物酶增殖物激活受体γ(PPARγ)、CCAAT-增强子结合蛋白β(C/EBPβ)、CCAAT-增强子结合蛋白α(C/EBPα)蛋白表达量。结果与空白组比较,模型组大鼠股骨脂肪细胞数目以及骨髓PPARγ、C/EBPβ、C/EBPα蛋白表达明显升高(P<0.01);与模型组比较,补佳乐组、左归丸组、右归丸组大鼠股骨脂肪细胞数目以及骨髓PPARγ、C/EBPβ、C/EBPα蛋白表达量均有不同程度降低,并且补佳乐组、左归丸组优于右归丸组(P<0.05或P<0.01)。结论左归丸与右归丸均能改善去卵巢大鼠骨髓微环境,其机制可能与抑制股骨脂肪分化和骨髓PPARγ、C/EBPβ、C/EBPα蛋白表达有关。
Objective To explore the possible mechanism of Zuogui Pill( 左归丸) and Yougui Pill( 右归丸) in treating postmenopausal osteoporosis( PMOP).Methods Sixty rats were randomized into a blank group,a sham operation group,a model group,an estradiol valerate tablets group,a Zuogui Pill group and a Yougui Pill group,10 rats in each group.Except the blank group and the sham operation group,rats in other groups were built PMOP models by bilateral oophorectomy.After 3 days,the Zuogui Pill group,Yougui Pill group and estradiol valerate tablets group were given corresponding medicine by gavage,with the dose of 9.45 g /( kg·d),10.26 g /( kg·d),and 0.09 g /( kg·d),respectively.The blank group,the sham operation group and the model group were given 2 ml of distilled water by gavage.All groups were treated for 12 weeks.After intragastric administration was over,morphological changes of bone in proximal femur was examined by hematoxylin and eosin stain( HE stain).The expressions of femur bone marrow peroxisome proliferator activated receptorsγ( PPARγ), CCAAT-enhancer binding proteinβ( C /EBPβ) and CCAAT-enhancer binding proteinα( C /EBPα) were detected with Western blot method.Results Compared with the blank group,fat cells in the femur,and the expressions of bone marrow PPARγ,C / EBPβ and C / EBPα proteins in the model group increased( P < 0.01).Compared with the model group,fat cells in the femur,and the expressions of bone marrow PPARγ,C / EBPβ and C / EBPα proteins in the estradiol valerate tablets group,the Zuogui Pill group and the Yougui Pill group decreased to varying degrees,especially in the estradiol valerate tablets group and the Zuogui Pill group( P < 0.05 or P < 0.01).Conclusion Zuogui Pill and Yougui Pill might both improve the bone marrow microenvironment of ovariectomized rats.The mechanism might relate to inhibiting the femur adipose differentiation and the expressions of bone marrow PPARγ,C / EBPβ and C / EBPαproteins.
Objective To explore the possible mechanism of Zuogui Pill( 左归丸) and Yougui Pill( 右归丸) in treating postmenopausal osteoporosis( PMOP).Methods Sixty rats were randomized into a blank group,a sham operation group,a model group,an estradiol valerate tablets group,a Zuogui Pill group and a Yougui Pill group,10 rats in each group.Except the blank group and the sham operation group,rats in other groups were built PMOP models by bilateral oophorectomy.After 3 days,the Zuogui Pill group,Yougui Pill group and estradiol valerate tablets group were given corresponding medicine by gavage,with the dose of 9.45 g /( kg·d),10.26 g /( kg·d),and 0.09 g /( kg·d),respectively.The blank group,the sham operation group and the model group were given 2 ml of distilled water by gavage.All groups were treated for 12 weeks.After intragastric administration was over,morphological changes of bone in proximal femur was examined by hematoxylin and eosin stain( HE stain).The expressions of femur bone marrow peroxisome proliferator activated receptorsγ( PPARγ), CCAAT-enhancer binding proteinβ( C /EBPβ) and CCAAT-enhancer binding proteinα( C /EBPα) were detected with Western blot method.Results Compared with the blank group,fat cells in the femur,and the expressions of bone marrow PPARγ,C / EBPβ and C / EBPα proteins in the model group increased( P < 0.01).Compared with the model group,fat cells in the femur,and the expressions of bone marrow PPARγ,C / EBPβ and C / EBPα proteins in the estradiol valerate tablets group,the Zuogui Pill group and the Yougui Pill group decreased to varying degrees,especially in the estradiol valerate tablets group and the Zuogui Pill group( P < 0.05 or P < 0.01).Conclusion Zuogui Pill and Yougui Pill might both improve the bone marrow microenvironment of ovariectomized rats.The mechanism might relate to inhibiting the femur adipose differentiation and the expressions of bone marrow PPARγ,C / EBPβ and C / EBPαproteins.
引文
[1]DALLE CARBONARE L,VALENTI MT,ZANATTA M,et al.Circulating mesenchymal stem cells with abnormal osteogenic differentiation in patients with osteoporosis[J].Arthritis Rheum,2009,60(11):3356-3365.
[2]李慧武,戴尅戎.骨髓内成骨与成脂平衡的调控[J].国外医学骨科学分册,2005,26(5):288-289.
[3]KUSHWAHA P,KHEDGIKAR V,GAUTAM J,et al.A novel therapeutic approach with Caviunin-based isoflavonoid that en routes bone marrow cells to bone formation via BMP2/Wnt-β-catenin signaling[J].Cell Death Dis,2014,5:e1422.doi:10.1038/cddis.2014.350.
[4]NUTTALL ME,GIMBLE JM.Controlling the balance between osteoblastogenesis and adipogenesis and the consequent therapeutic implications[J].Curr Opin Pharmacol,2004,4(3):290-294.
[5]FAN JZ,WANG Y,MENG Y,et al.Panax notoginseng saponins mitigate ovariectomy-induced bone loss and inhibit marrow adiposity in rats[J].Menopause,2015,22(12):1343-1350.
[6]YEUNG DK,GRIFFITH JF,ANTONIO GE,et al.Osteoporosis is associated with increased marrow fat content and decreased marrow fat unsaturation:a proton MR spectroscopy study[J].J Magn Reson Imaging,2005,22(2):279-285.
[7]MYERS TJ,GRANERO-MOLTO F,LONGOBARDI L,et al.Mesenchymal stem cells at the intersection of cell and gene therapy[J].Expert Opin Biol Ther,2010,10(12):1663-1679.
[8]孙月娇.左、右归丸及其拆方调节去卵巢骨质疏松大鼠成骨与成脂平衡的研究[D].沈阳:辽宁中医药大学,2015.
[9]李书娟.左、右归丸及其拆方调节去卵巢大鼠脂质代谢的研究[D].沈阳:辽宁中医药大学,2016.
[10]刑丽,解汝娟.过氧化物酶增殖物激活受体与骨代谢[J].中国老年学杂志,2007,27(17):1742-1744.
[11]KNOUFF C,AUWERX J.Peroxisome proliferator-activated receptor-gamma calls for activation in moderation:lessons from genetics and pharmacology[J].Endocr Rev,2004,25(6):899-918.
[12]CONZE D,WEISS L,REGEN PS,et al.Autocrine production of interleukin 6 causes multidrug resistance in breast cancer cells[J].Cancer Res,2001,61(24):8851-8858.
[13]蒋旭鹏,潘晓琳,潘泽民.转录因子C/EBPβ的研究进展[J].中国公共卫生,2011,27(12):1628-1629.
[14]HU E,TONTONOZ P,SPIEGELMAN BM.Transdifferentiation of myoblasts by the adipogenic transcription factors PPAR gamma and C/EBP alpha[J].Proc Natl Acad Sci USA,1995,92(21):9856-9860.
[15]ROSEN ED,WALKEY CJ,PUIGSERVER P,et al.Transcriptional regulation of adipogenesis[J].Genes Dev,2000,14(11):1293-1307.
[16]TANG QQ,LANE MD.Activation and centromeric localization of CCAAT/enhancer-binding proteins during the mitotic clonal expansion of adipocyte differentiation[J].Genes Dev,1999,13(17):2231-2241.
[2]李慧武,戴尅戎.骨髓内成骨与成脂平衡的调控[J].国外医学骨科学分册,2005,26(5):288-289.
[3]KUSHWAHA P,KHEDGIKAR V,GAUTAM J,et al.A novel therapeutic approach with Caviunin-based isoflavonoid that en routes bone marrow cells to bone formation via BMP2/Wnt-β-catenin signaling[J].Cell Death Dis,2014,5:e1422.doi:10.1038/cddis.2014.350.
[4]NUTTALL ME,GIMBLE JM.Controlling the balance between osteoblastogenesis and adipogenesis and the consequent therapeutic implications[J].Curr Opin Pharmacol,2004,4(3):290-294.
[5]FAN JZ,WANG Y,MENG Y,et al.Panax notoginseng saponins mitigate ovariectomy-induced bone loss and inhibit marrow adiposity in rats[J].Menopause,2015,22(12):1343-1350.
[6]YEUNG DK,GRIFFITH JF,ANTONIO GE,et al.Osteoporosis is associated with increased marrow fat content and decreased marrow fat unsaturation:a proton MR spectroscopy study[J].J Magn Reson Imaging,2005,22(2):279-285.
[7]MYERS TJ,GRANERO-MOLTO F,LONGOBARDI L,et al.Mesenchymal stem cells at the intersection of cell and gene therapy[J].Expert Opin Biol Ther,2010,10(12):1663-1679.
[8]孙月娇.左、右归丸及其拆方调节去卵巢骨质疏松大鼠成骨与成脂平衡的研究[D].沈阳:辽宁中医药大学,2015.
[9]李书娟.左、右归丸及其拆方调节去卵巢大鼠脂质代谢的研究[D].沈阳:辽宁中医药大学,2016.
[10]刑丽,解汝娟.过氧化物酶增殖物激活受体与骨代谢[J].中国老年学杂志,2007,27(17):1742-1744.
[11]KNOUFF C,AUWERX J.Peroxisome proliferator-activated receptor-gamma calls for activation in moderation:lessons from genetics and pharmacology[J].Endocr Rev,2004,25(6):899-918.
[12]CONZE D,WEISS L,REGEN PS,et al.Autocrine production of interleukin 6 causes multidrug resistance in breast cancer cells[J].Cancer Res,2001,61(24):8851-8858.
[13]蒋旭鹏,潘晓琳,潘泽民.转录因子C/EBPβ的研究进展[J].中国公共卫生,2011,27(12):1628-1629.
[14]HU E,TONTONOZ P,SPIEGELMAN BM.Transdifferentiation of myoblasts by the adipogenic transcription factors PPAR gamma and C/EBP alpha[J].Proc Natl Acad Sci USA,1995,92(21):9856-9860.
[15]ROSEN ED,WALKEY CJ,PUIGSERVER P,et al.Transcriptional regulation of adipogenesis[J].Genes Dev,2000,14(11):1293-1307.
[16]TANG QQ,LANE MD.Activation and centromeric localization of CCAAT/enhancer-binding proteins during the mitotic clonal expansion of adipocyte differentiation[J].Genes Dev,1999,13(17):2231-2241.