Article(id=1291704918201623107, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1291704786705993936, articleNumber=null, orderNo=null, doi=10.13699/j.cnki.1001-6821.2025.20.009, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1752508800000, receivedDateStr=2025-07-15, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1785896051215, onlineDateStr=2026-08-05, pubDate=1761580800000, pubDateStr=2025-10-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1785896051215, onlineIssueDateStr=2026-08-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1785896051215, creator=13701087609, updateTime=1785896051215, updator=13701087609, issue=Issue{id=1291704786705993936, tenantId=1146029695717560320, journalId=1246415772164075586, year='2025', volume='41', issue='20', pageStart='2851', pageEnd='3000', issueExtLink='null', onlineDate='null', pubDate='1761580800000', pubDateStr='2025-10-28', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1785896019864, creator='13701087609', updateTime=1785896741492, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1291707813508702571, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1291704786705993936, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1291707813508702572, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1291704786705993936, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=2899, endPage=2905, ext={EN=ArticleExt(id=1291704918444892740, articleId=1291704918201623107, tenantId=1146029695717560320, journalId=1246415772164075586, language=EN, title=Research on the effect of β-sitosterol on osteogenic differentiation of bone marrow mesenchymal stem cells by regulating the TGF-β1/Smad3 pathway, columnId=1246531407326105792, journalTitle=Chinese Journal of Clinical Pharmacology, columnName=Clinical and Basic Bridging Research, runingTitle=null, highlight=null, articleAbstract=
Objective

To investigate the effect of β-sitosterol on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by regulating the transforming growth factor β1 (TGF-β1)/mothers against decapentaplegic drosophila homolog 3 (Smad3) pathway.

Methods

BMSCs were randomly assigned into control group, low-dose β-sitosterol (β-sitosterol-L) group, medium-dose β-sitosterol group (β-sitosterol-M), and high-dose β-sitosterol group (β-sitosterol-H). Control group was cultured normally and β-sitosterol-L, -M and -H group was intervened by 1.5、3.0、6.0 μg·mL-1 β-sitosterol. BMSCs were randomly divided into control group (normal culture), β-sitosterol group (6 μg·mL-1 β-sitosterol), SIS3 group (5 μmol·L-1 SIS3), and β-sitosterol+SIS3 group (6 μg·mL-1 β-sitosterol+5 μmol·L-1 SIS3). Enzyme-linked immunosorbent assay (ELISA) was used to detect the alkaline phosphatase (ALP) activity was measured in each group. Alizarin red staining was used to determine the calcium deposition rate of each group, and then the osteogenic differentiation of BMSCs in each group was detected. Western blot was used to detect the osteogenic differentiation of BMSCs and the expression of TGF-β1/Smad3 pathway related proteins in each group.

Results

The ALP activities of the β-sitosterol-L, β-sitosterol-M, β-sitosterol-H and the control groups were (1.24±0.10), (1.53±0.08), (1.84±0.11) and (0.95±0.09) U·mg prot-1, respectively; the calcium deposition rates were (196.21±13.67)%, (301.73±16.90)%, (423.60±20.25)% and (100.00±0)%, respectively; the relative expression levels of bone morphogenetic protein 2 (BMP-2) protein were 0.49±0.05, 0.89±0.07, 1.30±0.09 and 0.13±0.02, respectively; the relative protein expression of TGF-β 1 were 0.47±0.04, 0.85±0.07, 1.26±0.09 and 0.10±0.01, respectively; the p-Smad3/Smad3 values were 0.32±0.03, 0.59±0.06, 0.94±0.05 and 0.08±0.02, respectively. Compared with the control group, the above indicators of the β-sitosterol-L, β-sitosterol-M and β-sitosterol-H groups showed an increase in a dose-dependent manner, and the differences were statistically significant (all P<0.05). The ALP activities of the β-sitosterol group, SIS3 group, β-sitosterol+SIS3 group and the control group were (1.78±0.14), (0.43±0.08), (0.97±0.13) and (0.91±0.10) U·mg prot-1, respectively; the calcium deposition rates were (410.35±19.72)%, (45.62±13.91)%, (109.14±16.53)% and (100.00±0)%, respectively; the relative expression levels of BMP-2 protein were 1.79±0.16, 0.27±0.07, 0.87±0.10 and 0.85±0.11, respectively; the relative protein expression levels of TGF-β1 were 1.47±0.10, 0.11±0.03, 0.56±0.09 and 0.55±0.06, respectively; The p-Smad3/Smad3 values were 0.94±0.06, 0.09±0.02, 0.45±0.05 and 0.43±0.04, respectively. Compared with the control group, the above indicators were all increased in the β-sitosterol group, while they were all decreased in the SIS3 group, and the differences were statistically significant (all P<0.05). Compared with the β-sitostero group, the above indicators were all decreased in the β-sitosterol+SIS3 group, and the differences were statistically significant (all P<0.05).

Conclusion

β-sitosterol can promote osteogenic differentiation of BMSCs by activating the TGF-β1/Smad3 pathway.

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目的

探讨β-谷甾醇调控转化生长因子β1(TGF-β1)/果蝇母体抗脑脊髓瘫痪同源物3(Smad3)通路对骨髓间充质干细胞(BMSCs)成骨分化的影响。

方法

将BMSCs随机分为对照组、β-谷甾醇低中剂量组、高剂量实验组,对照组正常培养,低、中、高剂量组分别以1.50、3.0、6.0 μg·mL-1的β-谷甾醇干预。将BMSCs随机分为对照组、β-谷甾醇组(6 μg·mL-1 β-谷甾醇)、Smad3抑制剂(SIS3)组(5 μmol·L-1 SIS3)、β-谷甾醇+SIS3组(6 μg·mL-1 β-谷甾醇+5μmol·L-1 SIS3)。以酶联免疫吸附试验(ELISA)法测定各组碱性磷酸酶(ALP)活性,以茜素红染色测定各组钙沉积率;以蛋白质印迹(Western blot)法检测各组BMSCs成骨分化与TGF-β1/Smad3通路相关蛋白表达。

结果

β-谷甾醇低、中、高剂量组和对照组的ALP活性分别为(1.24±0.10)、(1.53±0.08)、(1.84±0.11)和(0.95±0.09)U·mg prot-1;钙沉积率分别为(196.21±13.67)%、(301.73±16.90)%、(423.60±20.25)%和(100.00±0)%;骨形态发生蛋白2(BMP-2)蛋白相对表达水平分别为0.49±0.05、0.89±0.07、1.30±0.09和0.13±0.02;TGF-β1蛋白相对表达水平分别为0.47±0.04、0.85±0.07、1.26±0.09和0.10±0.01;p-Smad3/Smad3蛋白相对表达水平分别为0.32±0.03、0.59±0.06、0.94±0.05和0.08±0.02。β-谷甾醇低、中、高剂量组上述指标与对照组比较均显著升高,并呈剂量依赖性,在统计学上差异均有统计学意义(均P<0.05)。β-谷甾醇组、SIS3组、β-谷甾醇+SIS3组和对照组的ALP活性分别为(1.78±0.14)、(0.43±0.08)、(0.97±0.13)和(0.91±0.10)U·mg prot-1;钙沉积率分别为(410.35±19.72)%、(45.62±13.91)%、(109.14±16.53)%和(100.00±0)%;BMP-2蛋白相对表达水平分别为1.79±0.16、0.27±0.07、0.87±0.10和0.85±0.11;TGF-β1蛋白相对表达水平分别为1.47±0.10、0.11±0.03、0.56±0.09和0.55±0.06;p-Smad3/Smad3蛋白相对表达水平分别为0.94±0.06、0.09±0.02、0.45±0.05和0.43±0.04。β-谷甾醇组上述指标与对照组比较均显著升高,而SIS3组均显著降低,在统计学上差异有统计学意义(均P<0.05)。β-谷甾醇+SIS3组上述指标与β-谷甾醇组比较均显著降低,在统计学上差异均有统计学意义(P<0.05)。

结论

β-谷甾醇可通过激活TGF-β1/Smad3通路促进BMSCs成骨分化。

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田甜(1982-),女,主管药师,主要从事药学基础研究工作

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李伟华,主任药师 MP: 13127269969 E-mail:
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田甜(1982-),女,主管药师,主要从事药学基础研究工作

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β-谷甾醇调控TGF-β1/Smad3通路对骨髓间充质干细胞成骨分化影响的研究
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田甜 a , 李刚 b , 王磊 c , 李伟华 a
中国临床药理学杂志 | 临床与基础桥接研究 2025,41(20): 2899-2905
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中国临床药理学杂志 |临床与基础桥接研究 2025 , 41 (20) : 2899 -2905
β-谷甾醇调控TGF-β1/Smad3通路对骨髓间充质干细胞成骨分化影响的研究
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田甜a, 李刚b, 王磊c, 李伟华a
作者信息
  • a.济南市人民医院 药剂科 山东 济南 271100
  • b.济南市人民医院 科教科 山东 济南 271100
  • c.济南市人民医院 医务科 山东 济南 271100
通讯作者:
李伟华,主任药师 MP: 13127269969 E-mail:
作者简介:

田甜(1982-),女,主管药师,主要从事药学基础研究工作

Research on the effect of β-sitosterol on osteogenic differentiation of bone marrow mesenchymal stem cells by regulating the TGF-β1/Smad3 pathway
Tian TIANa, Gang LIb, Lei WANGc, Wei-hua LIa
Affiliations
  • a.Department of Pharmacy, Jinan People’s Hospital, Jinan 271100, Shandong Province, China
  • b.Department of Science and Education, Jinan People’s Hospital, Jinan 271100, Shandong Province, China
  • c.Department of Medical Affairs, Jinan People’s Hospital, Jinan 271100, Shandong Province, China
出版时间: 2025-10-28 doi: 10.13699/j.cnki.1001-6821.2025.20.009
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目的

探讨β-谷甾醇调控转化生长因子β1(TGF-β1)/果蝇母体抗脑脊髓瘫痪同源物3(Smad3)通路对骨髓间充质干细胞(BMSCs)成骨分化的影响。

方法

将BMSCs随机分为对照组、β-谷甾醇低中剂量组、高剂量实验组,对照组正常培养,低、中、高剂量组分别以1.50、3.0、6.0 μg·mL-1的β-谷甾醇干预。将BMSCs随机分为对照组、β-谷甾醇组(6 μg·mL-1 β-谷甾醇)、Smad3抑制剂(SIS3)组(5 μmol·L-1 SIS3)、β-谷甾醇+SIS3组(6 μg·mL-1 β-谷甾醇+5μmol·L-1 SIS3)。以酶联免疫吸附试验(ELISA)法测定各组碱性磷酸酶(ALP)活性,以茜素红染色测定各组钙沉积率;以蛋白质印迹(Western blot)法检测各组BMSCs成骨分化与TGF-β1/Smad3通路相关蛋白表达。

结果

β-谷甾醇低、中、高剂量组和对照组的ALP活性分别为(1.24±0.10)、(1.53±0.08)、(1.84±0.11)和(0.95±0.09)U·mg prot-1;钙沉积率分别为(196.21±13.67)%、(301.73±16.90)%、(423.60±20.25)%和(100.00±0)%;骨形态发生蛋白2(BMP-2)蛋白相对表达水平分别为0.49±0.05、0.89±0.07、1.30±0.09和0.13±0.02;TGF-β1蛋白相对表达水平分别为0.47±0.04、0.85±0.07、1.26±0.09和0.10±0.01;p-Smad3/Smad3蛋白相对表达水平分别为0.32±0.03、0.59±0.06、0.94±0.05和0.08±0.02。β-谷甾醇低、中、高剂量组上述指标与对照组比较均显著升高,并呈剂量依赖性,在统计学上差异均有统计学意义(均P<0.05)。β-谷甾醇组、SIS3组、β-谷甾醇+SIS3组和对照组的ALP活性分别为(1.78±0.14)、(0.43±0.08)、(0.97±0.13)和(0.91±0.10)U·mg prot-1;钙沉积率分别为(410.35±19.72)%、(45.62±13.91)%、(109.14±16.53)%和(100.00±0)%;BMP-2蛋白相对表达水平分别为1.79±0.16、0.27±0.07、0.87±0.10和0.85±0.11;TGF-β1蛋白相对表达水平分别为1.47±0.10、0.11±0.03、0.56±0.09和0.55±0.06;p-Smad3/Smad3蛋白相对表达水平分别为0.94±0.06、0.09±0.02、0.45±0.05和0.43±0.04。β-谷甾醇组上述指标与对照组比较均显著升高,而SIS3组均显著降低,在统计学上差异有统计学意义(均P<0.05)。β-谷甾醇+SIS3组上述指标与β-谷甾醇组比较均显著降低,在统计学上差异均有统计学意义(P<0.05)。

结论

β-谷甾醇可通过激活TGF-β1/Smad3通路促进BMSCs成骨分化。

β-谷甾醇  /  TGF-β1/Smad3  /  骨髓间充质干细胞  /  成骨分化
Objective

To investigate the effect of β-sitosterol on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by regulating the transforming growth factor β1 (TGF-β1)/mothers against decapentaplegic drosophila homolog 3 (Smad3) pathway.

Methods

BMSCs were randomly assigned into control group, low-dose β-sitosterol (β-sitosterol-L) group, medium-dose β-sitosterol group (β-sitosterol-M), and high-dose β-sitosterol group (β-sitosterol-H). Control group was cultured normally and β-sitosterol-L, -M and -H group was intervened by 1.5、3.0、6.0 μg·mL-1 β-sitosterol. BMSCs were randomly divided into control group (normal culture), β-sitosterol group (6 μg·mL-1 β-sitosterol), SIS3 group (5 μmol·L-1 SIS3), and β-sitosterol+SIS3 group (6 μg·mL-1 β-sitosterol+5 μmol·L-1 SIS3). Enzyme-linked immunosorbent assay (ELISA) was used to detect the alkaline phosphatase (ALP) activity was measured in each group. Alizarin red staining was used to determine the calcium deposition rate of each group, and then the osteogenic differentiation of BMSCs in each group was detected. Western blot was used to detect the osteogenic differentiation of BMSCs and the expression of TGF-β1/Smad3 pathway related proteins in each group.

Results

The ALP activities of the β-sitosterol-L, β-sitosterol-M, β-sitosterol-H and the control groups were (1.24±0.10), (1.53±0.08), (1.84±0.11) and (0.95±0.09) U·mg prot-1, respectively; the calcium deposition rates were (196.21±13.67)%, (301.73±16.90)%, (423.60±20.25)% and (100.00±0)%, respectively; the relative expression levels of bone morphogenetic protein 2 (BMP-2) protein were 0.49±0.05, 0.89±0.07, 1.30±0.09 and 0.13±0.02, respectively; the relative protein expression of TGF-β 1 were 0.47±0.04, 0.85±0.07, 1.26±0.09 and 0.10±0.01, respectively; the p-Smad3/Smad3 values were 0.32±0.03, 0.59±0.06, 0.94±0.05 and 0.08±0.02, respectively. Compared with the control group, the above indicators of the β-sitosterol-L, β-sitosterol-M and β-sitosterol-H groups showed an increase in a dose-dependent manner, and the differences were statistically significant (all P<0.05). The ALP activities of the β-sitosterol group, SIS3 group, β-sitosterol+SIS3 group and the control group were (1.78±0.14), (0.43±0.08), (0.97±0.13) and (0.91±0.10) U·mg prot-1, respectively; the calcium deposition rates were (410.35±19.72)%, (45.62±13.91)%, (109.14±16.53)% and (100.00±0)%, respectively; the relative expression levels of BMP-2 protein were 1.79±0.16, 0.27±0.07, 0.87±0.10 and 0.85±0.11, respectively; the relative protein expression levels of TGF-β1 were 1.47±0.10, 0.11±0.03, 0.56±0.09 and 0.55±0.06, respectively; The p-Smad3/Smad3 values were 0.94±0.06, 0.09±0.02, 0.45±0.05 and 0.43±0.04, respectively. Compared with the control group, the above indicators were all increased in the β-sitosterol group, while they were all decreased in the SIS3 group, and the differences were statistically significant (all P<0.05). Compared with the β-sitostero group, the above indicators were all decreased in the β-sitosterol+SIS3 group, and the differences were statistically significant (all P<0.05).

Conclusion

β-sitosterol can promote osteogenic differentiation of BMSCs by activating the TGF-β1/Smad3 pathway.

β-sitosterol  /  transforming growth factor β1/mothers against decapentaplegic drosophila homolog 3  /  bone marrow mesenchymal stem cells  /  osteogenic differentiation
田甜, 李刚, 王磊, 李伟华. β-谷甾醇调控TGF-β1/Smad3通路对骨髓间充质干细胞成骨分化影响的研究. 中国临床药理学杂志, 2025 , 41 (20) : 2899 -2905 . DOI: 10.13699/j.cnki.1001-6821.2025.20.009
Tian TIAN, Gang LI, Lei WANG, Wei-hua LI. Research on the effect of β-sitosterol on osteogenic differentiation of bone marrow mesenchymal stem cells by regulating the TGF-β1/Smad3 pathway[J]. Chinese Journal of Clinical Pharmacology, 2025 , 41 (20) : 2899 -2905 . DOI: 10.13699/j.cnki.1001-6821.2025.20.009
  • 山东省医药卫生科技发展计划基金资助项目(202002040822)
  • 济南市科技创新发展计划基金资助项目(202134050)
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doi: 10.13699/j.cnki.1001-6821.2025.20.009
  • 接收时间:2025-07-15
  • 首发时间:2026-08-05
  • 出版时间:2025-10-28
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  • 收稿日期:2025-07-15
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山东省医药卫生科技发展计划基金资助项目(202002040822)
济南市科技创新发展计划基金资助项目(202134050)
作者信息
    a.济南市人民医院 药剂科 山东 济南 271100
    b.济南市人民医院 科教科 山东 济南 271100
    c.济南市人民医院 医务科 山东 济南 271100

通讯作者:

李伟华,主任药师 MP: 13127269969 E-mail:
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
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栓菌属 Trametes 5 2.39
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