β-蜕皮甾酮对高糖诱导的大鼠成骨细胞增殖、分化和凋亡的影响  

Effects of β-ecdysterone on the proliferation, differentiation and apoptosis of rat osteoblasts induced by high glucose

作　　者：赵强 王字兴[3] 何江涛[2] ZHAO Qiang;WANG Zi-xing;HE Jiang-tao(Department of Clinical Medicine,North Sichuan Medical College,Nanchong 637000,Sichuan Province,China;Department of Orthopedics,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,Sichuan Province,China;Department of Orthopedics,Second Affiliated Hospital of North Sichuan Medical College,Nanchong 637100,Sichuan Province,China)

机构地区：[1]川北医学院临床医学系,四川南充637000 [2]川北医学院附属医院骨科,四川南充637000 [3]川北医学院第二附属医院骨科,四川南充637100

出　　处：《中国临床药理学杂志》2021年第4期443-446,共4页The Chinese Journal of Clinical Pharmacology

基　　金：四川省基层卫生事业发展研究中心科研基金资助项目(SWFZ19-Y-30)。

摘　　要：目的探讨β-蜕皮甾酮对高糖诱导的大鼠成骨细胞增殖、分化和凋亡的影响及分子机制。方法将大鼠成骨细胞随机分为空白组(5.5 mmol·L-1葡萄糖)、模型组(25 mmol·L-1葡萄糖)、低、中、高剂量实验组(1,5,25μmol·L-1β-蜕皮甾酮+25 mmol·L-1葡萄糖)、p38 MAPK激活组(10μmol·L-1 anisomycin+25μmol·L-1β-蜕皮甾酮+25 mmol·L-1葡萄糖)、阴性对照组(与anisomycin等量的PBS+25μmol·L-1β-蜕皮甾酮+25 mmol·L-1葡萄糖)。细胞计数试剂盒8(CCK-8)检测细胞活性;流式细胞术检测细胞凋亡情况;用蛋白质印迹法检测Run相关基因2(RunX2)、碱性磷酸酶(ALP)、骨钙素(OCN)和磷酸化p38丝裂原活化蛋白激酶(p-p38 MAPK)蛋白表达。结果空白组、模型组、低、中、高剂量实验组、阴性对照组、p38 MAPK激活组大鼠成骨细胞活性分别为(102.41±5.34)%,(68.57±5.25)%,(75.36±7.06)%,(79.13±8.24)%,(83.20±8.89)%,(83.17±8.93)%,(71.11±6.16)%;细胞凋亡率分别为(4.23±0.56)%,(18.57±1.96)%,(13.08±1.34)%,(10.30±0.95)%,(8.25±1.08)%,(8.36±1.12)%,(16.48±1.56)%;RunX2蛋白表达水平分别为0.81±0.07,0.31±0.04,0.45±0.06,0.55±0.04,0.61±0.08,0.63±0.07,0.41±0.05;ALP蛋白表达水平分别为0.75±0.08,0.38±0.05,0.49±0.07,0.59±0.05,0.65±0.08,0.68±0.07,0.44±0.06;OCN蛋白表达水平分别为0.69±0.05,0.28±0.03,0.34±0.05,0.47±0.04,0.45±0.05,0.46±0.05,0.31±0.04;p-p38 MAPK蛋白表达水平分别为0.22±0.04,0.84±0.09,0.59±0.06,0.41±0.04,0.32±0.04,0.32±0.07,0.74±0.07;模型组与空白组相比,低、中、高剂量实验组与模型组相比,p38 MAPK激活组与阴性对照组相比,差异均有统计学意义(均P<0.05)。结论β-蜕皮甾酮可能通过抑制p38 MAPK信号通路促进大鼠成骨细胞增殖、分化,抑制细胞凋亡。Objective To investigate the effect and molecular mechanism of β-ecdysterone on the proliferation, differentiation and apoptosis of rat osteoblasts induced by high glucose.Methods Rat osteoblasts were randomly divided into blank group(5.5 mmol·L-1 glucose), model group(25 mmol·L-1 glucose), test-L/-M/-H group(1, 5, 25 μmol·L-1 β-ecdysterone+25 mmol·L-1 glucose), p38 MAPK activation group(10 μmol·L-1 anisomycin+25 μmol·L-1 β-ecdysterone+25 mmol·L-1 glucose), negative control group (equivalent to the amount of anisomycin PBS + 25 μmol·L-1β-ecdysterone + 25 mmol·L-1 glucose).Cell counting kit 8(CCK-8) was used to detect cell activity;flow cytometry was used to detect cell apoptosis;Western blot was used to analysis Run-related gene 2(Run X2),alkaline phosphatase(ALP),osteocalcin(OCN) and phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK) protein expression.Results The activity of rat osteoblasts in the blank group,model group,test-L/-M/-H group,negative control group,p38 MAPK activation group were(102.41 ± 5.34) %,(68.57 ± 5.25) %,(75.36 ± 7.06) %,(79.13 ± 8.24) %,(83.20 ± 8.89) %,(83.17 ± 8.93) %,(71.11 ± 6.16) %, respectively;the apoptosis rates were(4.23 ± 0.56) %,(18.57 ± 1.96) %,(13.08 ± 1.34) %,(10.30 ± 0.95) %,(8.25 ± 1.08) %,(8.36 ± 1.12) %,(16.48 ± 1.56) %,respectively;Run X2 protein expression were 0.81 ± 0.07,0.31 ± 0.04,0.45 ± 0.06,0.55 ± 0.04,0.61 ± 0.08,0.63 ± 0.07,0.41 ± 0.05,respectively;ALP protein expressions were0.75 ± 0.08,0.38 ± 0.05,0.49 ± 0.07,0.59 ± 0.05,0.65 ± 0.08,0.68 ± 0.07,0.44 ± 0.06,respectively;OCN protein expressions were 0.69 ± 0.05,0.28 ± 0.03,0.34 ± 0.05,0.47 ± 0.04,0.45 ± 0.05,0.46 ± 0.05,0.31 ± 0.04,respectively;p-p38 MAPK protein expressions were 0.22 ± 0.04,0.84 ± 0.09,0.59 ± 0.06,0.41 ± 0.04,0.32 ± 0.04,0.32 ± 0.07,0.74 ± 0.07,respectively;Comparison between model group and blank group,comparison between test-L/-M/-H group and model group,comparison between p38 MAPK activation group and negative control g

关 键 词：β-蜕皮甾酮 p38丝裂原活化蛋白激酶信号通路 成骨细胞 增殖 分化 

分 类 号：R28[医药卫生—中药学]