[1]赵强,肖波,陈天逸,等.积雪草苷对脂多糖诱导的成骨细胞增殖和分化的影响[J].中国中医骨伤科杂志,2024,32(07):24-28+33.[doi:10.20085/j.cnki.issn1005-0205.240704 ]
　ZHAO Qiang,XIAO Bo,CHEN Tianyi,et al.Eficacy of Asiaticoside on Proliferation and Differentiation of Osteoblasts Induced by Lipopolysaccharide[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2024,32(07):24-28+33.[doi:10.20085/j.cnki.issn1005-0205.240704 ]
点击复制

积雪草苷对脂多糖诱导的成骨细胞增殖和分化的影响()
分享到：

《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第32卷
期数:
2024年07期
页码:
24-28+33
栏目:
实验研究
出版日期:
2024-07-15

文章信息/Info

Title:
Eficacy of Asiaticoside on Proliferation and Differentiation of Osteoblasts Induced by Lipopolysaccharide
文章编号:
1005-0205(2024)07-0024-05
作者:
赵强1; 肖波1; 陈天逸1; 赵明1; 杨国奇1; 徐银之1
1成都市郫都区人民医院(成都,611730)
Author(s):
ZHAO Qiang1; XIAO Bo1; CHEN Tianyi1; ZHAO Ming1; YANG Guoqi1; XU Yinzhi1
1Pidu District People's Hospital in Chengdu,Chengdu 611730,Cihna.
关键词:
积雪草苷;  脂多糖;  成骨细胞;  增殖;  分化
Keywords:
asiaticoside;  lipopolysaccharides;  osteoblasts;  proliferation;  differentiation
分类号:
R-33
DOI:
10.20085/j.cnki.issn1005-0205.240704
文献标志码:
A
摘要:
目的:探讨积雪草苷(AS)对脂多糖(LPS)诱导的MC3T3-E1成骨细胞增殖、分化的影响。方法:先用2 mg/L脂多糖处理MC3T3-E1成骨细胞,再用5,10,20,40,80 μmoL/L积雪草苷处理脂多糖诱导的MC3T3-E1成骨细胞,检测细胞活性,筛选最佳药物浓度; 将MC3T3-E1成骨细胞分为对照组(Control组)、脂多糖组(LPS组)、积雪草苷低、中、高浓度组(AS-L、AS-M、AS-H组)、积雪草苷高浓度+CXCR4抑制剂AMD3100组(AS-H+AMD3100组); Hoechst 33258染色观察细胞核形态; 碱性磷酸酶(ALP)染色及碱性磷酸酶活性检测细胞成骨能力; 茜素红染色法检测钙结节形成情况; 免疫印迹检测分化相关标志物Ⅰ型胶原(COL-Ⅰ)、骨桥蛋白(OPN)、骨钙素(OCN)及SDF-1、CXCR4表达情况。结果:筛选出10,20,40 μmoL/L积雪草苷为AS-L、AS-M、AS-H组药物浓度进行后续实验; 与对照组比较,脂多糖组MC3T3-E1成骨细胞细胞核固缩深染,呈明显细胞凋亡形态,碱性磷酸酶活性、钙结节形成、COL-Ⅰ、OPN、OCN、SDF-1、CXCR4蛋白表达显著下降(P<0.05); 与脂多糖组比较,AS-L、AS-M、AS-H组MC3T3-E1成骨细胞细胞核固缩深染逐渐减轻,凋亡形态细胞逐渐减少,碱性磷酸酶活性、钙结节形成、COL-Ⅰ、OPN、OCN、SDF-1、CXCR4蛋白表达依次显著上升(P<0.05); 与AS-H组比较,AS-H+AMD3100组MC3T3-E1成骨细胞细胞核固缩深染加重,凋亡形态细胞明显增多,碱性磷酸酶活性、钙结节形成、COL-Ⅰ、OPN、OCN、SDF-1、CXCR4蛋白表达显著下降(P<0.05)。结论:积雪草苷可通过激活SDF-1/CXCR4信号通路促进MC3T3-E1成骨细胞的增殖与分化。
Abstract:
Objective:To investigate the efficacy of asiaticoside(AS)on proliferation and differentiation of MC3T3-E1 osteoblasts induced by lipopolysaccharide(LPS).Methods:MC3T3-E1 osteoblasts were first treated with 2 mg/L LPS,and then LPS induced MC3T3-E1 osteoblasts were treated with 5,10,20,40,and 80 μmol/L asiaticoside.Cell activity was measured and the optimal drug concentration was selected.MC3T3-E1 osteoblasts were separated into control group,lipopolysaccharide group(LPS group),low,medium,and high concentration asiaticoside groups(AS-L,AS-M,AS-H group),and high concentration asiaticoside+CXCR4 inhibitor AMD3100 group(AS-H+AMD3100 group).Hoechst 33258 staining was applied to observe the morphology of the nucleus; alkaline phosphatase(ALP)staining and ALP activity were applied to detect the osteogenic ability of cells; alizarin red staining method was applied to detect the formation of calcium nodules; immunoblotting was applied to detect the expression of differentiation related biomarkers such as collagen type Ⅰ(COL-Ⅰ),osteopontin(OPN),osteocalcin(OCN),and SDF-1 and CXCR4.Results:10,20,and 40 μmol/L AS were selected as drug concentrations for AS-L,AS-M,and AS-H groups in subsequent experiments; compared with the control group,the nucleus of MC3T3-E1 osteoblasts in the LPS group showed pyknosis and deep staining,showing a obvious apoptotic morphology,the ALP activity,calcium nodule formation,the expression of COL-Ⅰ,OPN,OCN,SDF-1,CXCR4 proteins were greatly decreased(P<0.05).The nuclear pyknosis and deep staining of MC3T3-E1 osteoblasts in AS-L,AS-M,and AS-H groups gradually decreased compared with the LPS group,the number of apoptotic cells gradually decreased,the ALP activity,calcium nodule formation,the expression of COL-Ⅰ,OPN,OCN,SDF-1,CXCR4 proteins were obviously increased in sequence(P<0.05).The nuclei of MC3T3-E1 osteoblasts in the AS-H+AMD3100 group showed increased pyknosis and deep staining compared with the AS-H group,the apoptotic morphological cells obviously increased,the ALP activity,calcium nodule formation,the expression of COL-I,OPN,OCN,SDF-1,and CXCR4 proteins were greatly reduced(P<0.05).Conclusion:Asiaticoside can promote the proliferation and differentiation of MC3T3-E1 osteoblasts by activating SDF-1/CXCR4 signaling pathway.

参考文献/References:

[1] 谢梦生,龙燕鸣,李晓捷.慢病毒下调酪蛋白激酶2相互作用蛋白1表达的骨髓间充质干细胞修复骨质疏松大鼠牙槽骨缺损[J].中国组织工程研究,2023,27(10):1528-1533.
[2] BALDWIN P,LI D J,AUSTON D A,et al.Autograft,allograft,and bone graft substitutes:clinical evidence and indications for use in the setting of orthopaedic trauma surgery[J].J Orthop Trauma,2019,33(4):203-213.
[3] 曹振宇,沈晓钟,马建武,等.积雪草苷通过TGF-β/Smads通路对BMSCs成骨分化的影响[J].中国骨质疏松杂志,2022,28(5):695-700.
[4] 白书林,王奕佩,金珂,等.SDF-1α/CXCR4信号轴介导柚皮素对骨髓间充质干细胞成骨分化的影响[J].上海口腔医学,2021,30(6):579-584.
[5] HUANG Y,JIA Z,XU Y,et al.Selenium protects against LPS-induced MC3T3-E1 cells apoptosis through modulation of microRNA-155 and PI3K/Akt signaling pathways[J].Genet Mol Biol,2020,43(3):e20190153.
[6] 王辉,徐亮亮,孙大明,等.极度低氧条件下基质细胞衍生因子-1α过表达对间充质干细胞迁移的影响[J].中国医药导报,2022,19(16):5-10.
[7] TANG G,LIU Z,LIU Y,et al.Recent trends in the development of bone regenerative biomaterials[J].Front Cell Dev Biol,2021,9(1):665813-665830.
[8] LV J,SHARMA A,ZHANG T,et al.Pharmacological review on asiatic acid and its derivatives:a potential compound[J].SLAS Technol,2018,23(2):111-127.
[9] 丁平,张岱阳,李贺伟,等.积雪草苷对膝骨关节炎大鼠软骨损伤的影响及机制研究[J].中国中医骨伤科杂志,2022,30(9):7-13.
[10] 付涟桥,郭焱雄,李坚,等.积雪草苷调控miR-342-5p/AQP3轴保护IL-1β诱导的软骨细胞损伤[J].实验动物科学,2020,37(3):10-18.
[11] HE L,HONG G,ZHOU L,et al.Asiaticoside,a component of Centella asiatica attenuates RANKL-induced osteoclastogenesis via NFATc1 and NF-κB signaling pathways[J].J Cell Physiol,2019,234(4):4267-4276.
[12] 赵有顺,林平,涂迎春,等.RUNX2基因过表达载体修饰BMSC来源外泌体联合碳酸钙支架系统在骨缺损中的应用[J].中国骨伤,2022,35(4):379-386.
[13] 张好,王翠竹,皇甫慧敏,等.20(S)-原人参二醇对大鼠骨髓间充质干细胞成骨分化的影响及其机制[J].吉林大学学报(医学版),2023,49(4):867-874.
[14] 钱虹静,刘钰莲,张云鹏,等.淫羊藿苷对MC3T3-E1增殖及成骨分化的影响研究[J].口腔颌面外科杂志,2023,33(4):219-223.
[15] AI Y,SHE W,WU S,et al.AM1241-loaded poly(ethylene glycol)-dithiothreitol hydrogel repairs cranial bone defects by promoting vascular endothelial growth factor and COL-1 expression[J].Front Cell Dev Biol,2022,10(1):888598-888606.
[16] 张昕,李长顺,刘浩,等.人参皂苷Rg1促进人牙龈成纤维细胞增殖、迁移及成骨分化的研究[J].安徽医科大学学报,2023,58(5):812-819.
[17] 罗劲涛,王仕迎,韦标方.三七总皂苷对MC3T3-E1细胞增殖和成骨分化的影响[J].中成药,2023,45(7):2353-2358.
[18] 黄钟炼,罗绍伟,林海明,等.SDF-1/CXCR4介导体外冲击波促进大鼠骨髓间充质干细胞的黏附、迁移作用[J].颈腰痛杂志,2019,40(6):721-726.

更新日期/Last Update: 2024-07-05