健脾清肠补肾方药物血清对破骨细胞凋亡途径的影响  

Effects of Jianpi Qingchang Bushen Prescription on Apoptosis of Osteoclasts

作　　者：张亚利[1] 陈倩 戴彦成[1,2] 张紫薇 唐志鹏 ZHANG Yali;CHEN Qian;DAI Yancheng;ZHANG Ziwei;TANG Zhipeng(Longhua Hospital,Institute of Digestive Disease,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Shanghai Traditional Chinese Medicine-integrated Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200082,China)

机构地区：[1]上海中医药大学附属龙华医院,上海中医药大学脾胃病研究所,上海200032 [2]上海中医药大学附属上海市中西医结合医院,上海200082

出　　处：《中国中医药信息杂志》2021年第3期52-56,共5页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(81704009、81873253)。

摘　　要：目的观察健脾清肠补肾方药物血清对成熟破骨细胞凋亡的影响,探讨其可能作用机制。方法通过100 ng/mL核因子-κB受体活化因子配基(RANKL)和10 ng/mL巨噬细胞集落刺激因子(M-CSF)共同诱导小鼠单核细胞RAW264.7细胞株分化为破骨细胞,采用TRAP特异性染色进行鉴定。制备低、中、高剂量健脾清肠补肾方药物血清,终浓度分别为2.5%、5%、10%健脾清肠补肾方。流式细胞术检测破骨细胞凋亡率,Western blot检测破骨细胞Bcl-2、Bax蛋白的表达。结果采用RANKL和M-CSF可成功诱导RAW264.7细胞株分化为破骨细胞。与空白对照组比较,健脾清肠补肾方各剂量组细胞凋亡率均明显升高(P<0.05),破骨细胞Bcl-2蛋白表达明显下调、Bax蛋白表达明显上调,Bcl-2/Bax比值明显降低,其中健脾清肠补肾方高剂量组变化最明显。结论健脾清肠补肾方药物血清可促进破骨细胞凋亡,其作用机制可能是下调Bcl-2和上调Bax蛋白的表达。Objective To investigate the effects of Jianpi Qingchang Bushen Prescription on promoting the apoptosis of osteoclasts;To explore its possible mechanism of action.Methods RAW264.7 cells were induced to differentiate into osteoclasts by 100 ng/mL RNAKL and 10 ng/mL M-CSF.The induced cells were identified using TRAP staining.Medicated serum with Jianpi Qingchang Bushen Prescription was prepared.Three final concentrations of low,medium and high dosages(2.5%,5%,10%)of Jianpi Qingchang Bushen Prescription were formulated.Flow cytometry was used to detect the apoptosis rate of osteoclasts;Western blot was used to detect the expression of Bcl-2 and Bax proteins in osteoclasts.Results Mature osteoclasts were successfully induced by 100 ng/mL RNAKL and 10 ng/mL M-CSF.Compared with control group,cell apoptosis of the three Jianpi Qingchang Bushen Prescription groups increased(P<0.05),which significantly down-regulated Bcl-2 related to osteoclast apoptosis and up-regulate the expression of Bax protein,and the Bcl-2/Bax ratio was significantly reduced.Among them,Jianpi Qingchang Bushen Prescription high-dosages group changed the most.Conclusion Jianpi Qingchang Bushen Prescription can promote osteoclast apoptosis,and its mechanism may be achieved by down-regulating Bcl-2 and up-regulating the expression of Bax protein.

关 键 词：健脾清肠补肾方 细胞凋亡 破骨细胞 大鼠 

分 类 号：R285.5[医药卫生—中药学]