摘要
目的 探究黄芪多糖对MC-3T3-E1成骨细胞增殖及CYP17A1、CYP19A1基因与蛋白表达情况的影响,初步探讨其治疗糖皮质激素骨质疏松症(GIOP)的作用机制。方法 以MC-3T3-E1成骨细胞为研究对象分为6组,分别为正常组,模型组,维生素D_(3)组(1×10^(-5) mmol·L^(-1)),黄芪多糖高、中、低剂量组(1、0.1、0.01mg·L^(-1))。MTT法检测黄芪多糖对地塞米松诱导MC-3T3-E1成骨细胞增殖抑制率的影响;比色法检测黄芪多糖对地塞米松诱导MC-3T3-E1成骨细胞上清液中钙含量的影响;ELISA法检测黄芪多糖对地塞米松诱导MC-3T3-E1成骨细胞上清液25(OH)D_(3)含量的影响;RT-PCR和Western blot法分别检测黄芪多糖对地塞米松诱导MC-3T3-E1成骨细胞CYP17A1、CYP19A1 mRNA和蛋白相对表达水平的影响。结果 MTT检测结果显示,与正常组比较,模型组MC-3T3-E1成骨细胞在24 h、36 h、48 h时增殖抑制率显著升高(P<0.01);与模型组比较,黄芪多糖高、中、低剂量组及维生素D3组MC-3T3-E1成骨细胞在24 h、36 h、48 h增殖抑制率均显著降低(P<0.01)。比色法检测结果显示,与正常组比较,模型组MC-3T3-E1成骨细胞钙含量显著下降(P<0.01);与模型组比较,黄芪多糖高剂量组MC-3T3-E1成骨细胞钙含量显著升高(P<0.01)。ELISA检测结果显示,与正常组比较,模型组MC-3T3-E1成骨细胞25(OH)D_(3)含量显著下降(P<0.01);与模型组比较,黄芪多糖高、中剂量组MC-3T3-E1成骨细胞25(OH)D_(3)含量显著升高(P<0.01)。RT-PCR,Western-Blot检测结果显示,与正常组比较,模型组MC-3T3-E1成骨细胞中CYP17A1、CYP19A1 mRNA和蛋白相对表达量显著下降(P<0.01);与模型组比较,黄芪多糖高剂量组MC-3T3-E1成骨细胞中CYP17A1、CYP19A1 mRNA和蛋白相对表达量显著升高(P<0.01),并与药物浓度呈正相关。结论 黄芪多糖可改善糖皮质激素骨质疏松症相关指标,其可能机制与提高MC-3T3-E1成骨细胞活性和CYP17A1、CYP19A1 mRNA和蛋白表达水平有关。
Objective To investigate the effects of Astragalus polysaccharide on the proliferation of MC-3T3-E1 osteoblasts and the expression of CYP17A1 and CYP19A1 genes and proteins,and to explore the mechanism of Astragalus polysaccharide in the treatment of glucocorticoid osteoporosis(GIOP).Methods MC-3T3-E1 osteoblasts were studied and divided into 6 groups:normal group,model group,vitamin D_(3) group(1×10-5 mmol·L^(-1)),and Astragalus polysaccharide high-dose,medi⁃um-dose and low-dose groups(1,0.1,0.01mg·L^(-1)).The effect of Astragalus polysaccharide on the proliferation inhibition rate of MC-3T3-E1 osteoblasts induced by dexamethasone was determined by MTT method.The effect of Astragalus polysac⁃charide on calcium content in dexamethasone-induced supernatant of MC-3T3-E1 osteoblasts was determined by colorimetry.The effect of Astragalus polysaccharide on the content of 25(OH)D_(3) in the supernatant of MC-3T3-E1 osteoblasts induced by dexamethasone was determined by ELISA.The effects of Astragalus polysaccharide on the relative expression levels of CYP17A1 and CYP19A1 mRNA and protein in dexamethasone-induced MC-3T3-E1 osteoblasts were detected by RT-PCR and West⁃ern blot.Results Compared with the normal group,the proliferation inhibition rate of MC-3T3-E1 osteoblasts in the model group was significantly increased at 24,36,48 h(P<0.01).Compared with model group,the proliferation inhibition rate of MC-3T3-E1 osteoblasts in Astragalus polysaccharide high,medium and low dose groups and vitamin D_(3) groups was significant⁃ly decreased at 24,36,48 h(P<0.01).Colorimetric test results showed that calcium content of MC-3T3-E1 osteoblasts in model group was significantly decreased compared with normal group(P<0.01).Compared with model group,calcium content of MC-3T3-E1 osteoblasts in APS high-dose group was significantly increased(P<0.01).ELISA results showed that com⁃pared with normal group,the content of MC-3T3-E1 osteoblast 25(OH)D_(3) in model group was significantly decreased(P<0.01).Compared with model gr
作者
冯玘
何亚琼
谢小雪
龚华乾
李倩
李文
吴姣月
陈云志
柴艺汇
吴大梅
FENG Qi;HE Ya-qiong;XIE Xiao-xue;GONG Hua-qian;LI Qian;LI Wen;WU Jiao-yue;CHEN Yun-zhi;CHAI Yi-hui;WU Da-mei(North Anhui Health Vocational College,Suzhou,Anhui,234000,China;Guizhou University of Tradi-tional Chinese Medicine,Guiyang,Guizhou,550025,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2024年第1期97-101,共5页
Lishizhen Medicine and Materia Medica Research
基金
安徽高校自然科学研究重点项目(2022AH053142)
2021年国家级大学生创新创业训练计划项目(202110662012)。
