[1]颜国标,宋浩民,林南星△.薯蓣皂苷通过信号通路对大鼠骨髓间充质干细胞成骨分化的影响[J].中国中医骨伤科杂志,2023,31(07):1-5.[doi:10.20085/j.cnki.issn1005-0205.230701 ]
 YAN Guobiao,SONG Haomin,LIN Nanxing.Effect of Dioscin on Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells by Hedgehog Signaling Pathway[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2023,31(07):1-5.[doi:10.20085/j.cnki.issn1005-0205.230701 ]
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薯蓣皂苷通过信号通路对大鼠骨髓间充质干细胞成骨分化的影响()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第31卷
期数:
2023年07期
页码:
1-5
栏目:
实验研究
出版日期:
2023-07-15

文章信息/Info

Title:
Effect of Dioscin on Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells by Hedgehog Signaling Pathway
文章编号:
1005-0205(2023)07-0001-05
作者:
颜国标1宋浩民1林南星1△
1中国中医科学院望京医院药学部(北京,100102)
Author(s):
YAN Guobiao1SONG Haomin1LIN Nanxing1△
1Department of Pharmacy,Wangjing Hospital of China Academy of Chinese Medical Sciences,Beijing 100102,China.
关键词:
薯蓣皂苷 骨髓间充质干细胞 信号通路 成骨分化
Keywords:
dioscin bone marrow mesenchymal stem cells Hedgehog signaling pathway osteogenic differentiation
分类号:
R-33
DOI:
10.20085/j.cnki.issn1005-0205.230701
文献标志码:
A
摘要:
目的:探讨薯蓣皂苷对大鼠骨髓间充质干细胞(BMSCs)成骨分化及Hedgehog信号通路的影响。方法:分离培养大鼠骨髓间充质干细胞,分为对照组、薯蓣皂苷低(15%胎牛血清的DMEM培养基中加1 μmol/mL薯蓣皂苷)、中(15%胎牛血清的DMEM培养基中加3 μmol/mL薯蓣皂苷)、高(15%胎牛血清的DMEM培养基中加9 μmol/mL薯蓣皂苷)浓度组、阳性对照组(15%胎牛血清的DMEM培养基中加成骨诱导液),培养7 d后,茜素红染色观察骨髓间充质干细胞矿化结节; 细胞计数试剂盒-8法检测骨髓间充质干细胞增殖能力; 检测碱性磷酸酶(ALP)水平; 荧光定量PCR法检测骨髓间充质干细胞中SHH、IHH、Runx2信使RNA(mRNA)表达水平; 蛋白免疫印迹法检测骨髓间充质干细胞中Smo、Gli1、Ptch1蛋白表达水平。结果:对照组可见点状矿化结节,与对照组相比,薯蓣皂苷低、中、高浓度组的矿化结节数目依次增多,体积依次增大,骨髓间充质干细胞增殖率、碱性磷酸酶水平,SHH、IHH、Runx2 mRNA,Smo、Gli1、Ptch1蛋白表达水平依次升高,差异有统计学意义(P<0.05); 阳性对照组的矿化结节数目最多,连接成片,体积较大,且骨髓间充质干细胞增殖率、碱性磷酸酶水平,SHH、IHH、Runx2 mRNA,Smo、Gli1、Ptch1蛋白表达水平高于薯蓣皂苷高浓度组,差异有统计学意义(P<0.05)。结论:薯蓣皂苷可以促进大鼠骨髓间充质干细胞成骨分化,可能与Hedgehog信号通路的激活有关。
Abstract:
Objective:To explore the effect of dioscin on osteogenic differentiation and Hedgehog signaling pathway of rat bone marrow mesenchymal stem cells(BMSCs).Methods:Rat BMSCs were isolated and cultured,and divided into control group,dioscin low(15% fetal bovine serum DMEM medium supplemented with 1 μmol/mL dioscin),medium(15% fetal bovine serum DMEM medium supplemented with 3 μmol/mL dioscin),high(15% fetal bovine serum DMEM medium supplemented with 9 μmol/mL dioscin)concentration groups and positive control group(DMEM medium with 15% fetal bovine serum was supplemented with osteogenic induction solution).After 7 d of culture,the mineralized nodules of BMSCs were observed by Alizarin red staining,and Cell Count Kit-8 assay was used to detect the proliferation of BMSCs.Alkaline phosphatase(ALP)detection kit was used to detect ALP level.The mRNA expression levels of SHH,IHH,and Runx2 in BMSCs were detected by real-time PCR.The protein expression levels of Smo,Gli1,and Ptch1 in BMSCs were detected by Western Blot.Results:Punctate mineralized nodules were observed in the control group.Compared with the control group,the number and volume of mineralized nodules in the dioscin low,medium,high concentration groups were increased in sequence,and the proliferation rate,ALP level,SHH,IHH,Runx2 mRNA,Smo,Gli1,Ptch1 protein expression levels of BMSCs were increased in sequence(P<0.05).The positive control group had the most mineralized nodules,which connected into slices and had a large volume,and the proliferation rate,ALP level,SHH,IHH,Runx2 mRNA,Smo,Gli1,Ptch1 protein expression levels of BMSCs were higher than those in the dioscin high concentration group(P<0.05).Conclusion:Dioscin can promote the osteogenic differentiation of BMSCs in rats,which may be related to the activation of Hedgehog signaling pathway.

参考文献/References:

[1] PALACIOS S.Medical treatment of osteoporosis[J].Climacteric,2022,25(1):43-49.
[2] ZHANG Y,JIANG Y,LUO Y,et al.Interference of miR-212 and miR-384 promotes osteogenic differentiation via targeting RUNX2 in osteoporosis[J].Exp Mol Pathol,2020,113(1):104366-104367.
[3] AL-AZAB M,WALANA W,WEI J,et al.TL1A/TNFR2 axis enhances immunoregulatory effects of bone marrow derived mesenchymal stem cell by indian hedgehog signaling pathway[J].Int J Stem Cells,2021,14(1):58-73.
[4] AL-AZAB M,WANGB,ELKHIDER A,et al.Indian Hedgehog regulates senescence in bone marrow-derived mesenchymal stem cell through modulation of ROS/mTOR/4EBP1,p70S6K1/2 pathway[J].Aging(Albany NY),2020,12(7):5693-5715.
[5] BANDOPADHYAY S,ANAND U,GADEKAR V S,et al.Dioscin:a review on pharmacological properties and therapeutic values[J].Biofactors,2022,48(1):22-55.
[6] YIN W,LIU S,DONG M,et al.A new NLRP3 inflammasome inhibitor,dioscin,promotes osteogenesis[J].Small,2020,16(1):1905977-1905978.
[7] 陈学武,王弘.薯蓣皂苷对破骨细胞分化及骨吸收功能的调控作用及机制研究[J].中国临床药理学杂志,2022,38(6):523-527.
[8] 赖满香,廖利平,林基伟,等.梓醇调控Hedgehog信号通路促进BMSCs向成骨分化的实验研究[J].免疫学杂志,2019,35(11):921-926.
[9] REID I R.A broader strategy for osteoporosis interventions[J].Nat Rev Endocrinol,2020,16(6):333-339.
[10] QIU M,ZHAI S,FU Q,et al.Bone marrow mesenchymal stem cells-derived exosomal microrna-150-3p promotes osteoblast proliferation and differentiation in osteoporosis[J].Hum Gene Ther,2021,32(13):717-729.
[11] 周琦,孙慧娟,刘树民.穿山龙总皂苷调控巨噬细胞M1/M2极化治疗痛风性关节炎的作用机制[J].中国实验方剂学杂志,2021,27(24):92-99.
[12] ZHANG L,NI R,LI J,et al.Dioscin regulating bone marrow apoptosis in aplastic anemia[J].Drug Des Devel Ther,2022,16(1):3041-3053.
[13] 李菁,陈盈,滕伟,等.载基因脂多糖胺纳米囊泡诱导BMSCs定向成骨分化[J].中国修复重建外科杂志,2018,32(11):1469-1476.
[14] QI X,LI X.Mechanistic insights into the generation and transduction of hedgehog signaling[J].Trends Biochem Sci,2020,45(5):397-410.
[15] TAKEBE H,SHALEHIN N,HOSOYA A,et al.Sonic Hedgehog regulates bone fracture healing[J].Int J Mol Sci,2020,21(2):677-678.
[16] CHEN L,LIU G,LI W,et al.Synergistic effects of Indian Hedgehog and sonic Hedgehog on chondrogenesis during cartilage repair[J].J Mol Histol,2021,52(2):407-418.
[17] JIA Y,LU T,CHEN Q,et al.Exosomes secreted from sonic Hedgehog-modified bone mesenchymal stem cells facilitate the repair of rat spinal cord injuries[J].Acta Neurochir(Wien),2021,163(8):2297-2306.
[18] LIU D D,ZHANG C Y,LIU Y,et al.RUNX2 regulates osteoblast differentiation via the BMP4 signaling pathway[J].J Dent Res,2022,101(10):1227-1237.
[19] KOMORI T.Molecular mechanism of Runx2-dependent bone development[J].Mol Cells,2020,43(2):168-175.
[20] 程英雄,罗毅文,王斌,等.补肾活血汤调控Runx2及Osterix促进BMSCs成骨分化的作用研究[J].中国中医基础医学杂志,2018,24(8):1085-1088.
[21] RADHAKRISHNAN A,ROHATGI R,SIEBOLD C.Cholesterol access in cellular membranes controls Hedgehog signaling[J].Nat Chem Biol,2020,16(12):1303-1313.
[22] SHENG M,LIN Y,XU D,et al.CD47-mediated Hedgehog/SMO/GLI1 signaling promotes mesenchymal stem cell immunomodulation in mouse liver inflammation[J].Hepatology,2021,74(3):1560-1577.
[23] ZHONG L N,ZHANG Y Z,LI H,et al.Overexpressed miR-196a accelerates osteogenic differentiation in osteoporotic mice via GNAS-dependent Hedgehog signaling pathway[J].J Cell Biochem,2019,120(12):19422-19431.

备注/Memo

备注/Memo:
基金项目:北京市科学技术委员会科研计划项目(D201300006317016)
通信作者 E-mail:Nanson_lin@126.com
更新日期/Last Update: 2023-07-10