[1]余光书△,林焱斌,王松清,等.补骨颗粒含药血清对软骨细胞凋亡及缺氧诱导因子-1α/血管内皮生长因子-A信号通路的影响[J].中国中医骨伤科杂志,2023,31(04):1-8.[doi:10.20085/j.cnki.issn1005-0205.230401 ]
　YU Guangshu,LIN Yanbin,WANG Songqing,et al.Effects of Bugu Granules in Drug-Containing Serum on Chondrocyte Apoptosis and HIF-1α/VEGF-A Signaling Pathway[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2023,31(04):1-8.[doi:10.20085/j.cnki.issn1005-0205.230401 ]
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补骨颗粒含药血清对软骨细胞凋亡及缺氧诱导因子-1α/血管内皮生长因子-A信号通路的影响()
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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04期
页码:
1-8
栏目:
实验研究
出版日期:
2023-04-15

文章信息/Info

Title:
Effects of Bugu Granules in Drug-Containing Serum on Chondrocyte Apoptosis and HIF-1α/VEGF-A Signaling Pathway
文章编号:
1005-0205(2023)04-0001-08
作者:
余光书1△; 林焱斌2; 王松清1; 吴春玲1
1福州市第二医院疼痛科(福州,350007) 2福州市第二医院骨科
Author(s):
YU Guangshu1△; LIN Yanbin2; WANG Songqing1; WU Chunling1
1Department of Pains,Fuzhou Second Hospital,Fuzhou 350007,China; 2Department of Orthopaedics,Fuzhou Second Hospital,Fuzhou 350007,China.
关键词:
含药血清;  白细胞介素-1β;  软骨细胞;  凋亡;  缺氧诱导因子-1α/血管内皮生长因子-A信号通路
Keywords:
drug-containing serum;  interleukin-1β(IL-1β);  chondrocytes;  apoptosis;  HIF-1α/VEGF-A signaling pathway
分类号:
R-33
DOI:
10.20085/j.cnki.issn1005-0205.230401
文献标志码:
A
摘要:
目的:观察补骨颗粒含药血清对IL-1β诱导的软骨细胞凋亡及HIF-1α/VEGF-A信号通路的影响,探讨补骨颗粒对软骨细胞凋亡的影响机制。方法:通过体外培养大鼠膝关节软骨细胞,应用Cell Counting Kit-8 方法检测不同浓度补骨颗粒含药血清对软骨细胞活性的影响。将实验分为正常培养基组及不同浓度补骨颗粒含药血清(浓度为0%,5%,10%,15%,20%,25%,30%)联合10 ng/mL IL-1β培养组。应用膜联蛋白 V-异硫氰酸荧光素/碘化丙啶染色后经流式细胞仪检测软骨细胞凋亡影响情况,用探针(2,7-二氯二氢荧光素二乙酸酯)测量软骨细胞内活性氧的相对量,应用酶联免疫吸附试验检测软骨细胞HIF-1α及VEGF-A含量,应用逆转录酶定量聚合酶链式反应检测HIF-1α及VEGF-A基因表达情况。结果:浓度为5%,10%,15%补骨颗粒含药血清间的光密度(OD)值差异无统计学意义(P>0.05),而15%含药血清干预后软骨细胞的OD值比20%含药血清干预组(P=0.007)、25%含药血清干预组(P=0.003)及30%含药血清干预组(P<0.001)高,差异有统计学意义。应用10 ng/mL IL-1β干预后软骨细胞凋亡率和活性氧含量明显高于正常培养基组,并且15%含药血清对IL-1β诱导的软骨细胞凋亡的抑制率和软骨细胞中活性氧的含量高于其他浓度含药血清组,差异有统计学意义(P<0.05)。酶联免疫吸附试验检测结果表明15%和20%含药血清组HIF-1α和VEGF-A的含量差异无统计学意义(P=0.635,P=0.057); 而15%和20%含药血清组的HIF-1α和VEGF-A含量显著高于正常培养基组,差异有统计学意义(P<0.05); 而显著低于0%,5%,10%,25%和30%含药血清组,差异有统计学意义(P<0.01)。逆转录酶定量聚合酶链式反应检测结果表明15%含药血清组HIF-1α和VEGF-A基因表达量显著低于0%,5%和10%含药血清组,差异有统计学意义(P<0.05); 而15%含药血清组与20%,25%和30%含药血清组之间差异无统计学意义(P>0.05)。结论:补骨颗粒含药血清可以通过激活HIF-1α/VEGF-A信号通路而抑制IL-1β诱导的软骨细胞凋亡,并且含药血清浓度对软骨细胞的活性存在影响。
Abstract:
Objective:To observe the effects of Bugu granules in drug-containing serum on IL-1β-induced chondrocyte apoptosis and the expression of HIF-1α/VEGF-A signaling pathway-associated proteins,so as to explore the mechanism of action of Bugu granules on chondrocyte apoptosis.Methods:The chondrocytes were cultured in vitro,and cell counting Kit-8 was used to detect the effects of different concentrations of Bugu granule-containing serum on the activity of chondrocytes.Cells were divided to eight groups: a normal medium group and groups with different concentrations of Bugu granule-containing serum(concentrations of 0%,5%,10%,15%,20%,25%,30%)combined with 10 ng/mL IL-1β culture.The apoptosis of chondrocytes was detected by flow cytometry after annexin V-fluorescein isothiocyanate/propidium iodide staining.The relative amount of reactive oxygen species in chondrocytes was measured with a probe(2,7-dichlorodihydrofluorescein diacetate).The contents of HIF-1α and VEGF-A in chondrocytes were detected by enzyme-linked immunosorbent assay.The expression of HIF-1α and VEGF-A genes was detected by reverse-transcription quantitative polymerase chain reaction.Results:There were no significant differences in the OD value among the 0%,5%,10% and 15% drug-containing serum groups(P>0.05),while the OD value of chondrocytes after 15% drug-containing serum intervention was higher than that in the 20%,25% and 30% drug-containing serum groups(P=0.007,P=0.003 and P<0.001,respectively).The apoptosis rate of chondrocytes and content of reactive oxygen species in chondrocytes after 10 ng/mL IL-1β intervention was significantly higher than that in the normal medium group(P<0.001),and the inhibition rate of IL-1β-induced chondrocyte apoptosis and the content of reactive oxygen species in chondrocytes with 15% drug-containing serum was higher than those with the other concentrations of drug-containing serum(P<0.05).It was found that there were no significant differences in the contents of HIF-1α and VEGF-A between the 15% and 20% drug-containing serum groups(P=0.635 and P=0.057,respectively).The contents of HIF-1α and VEGF-A in the 15% and the 20% drug-containing serum groups were significantly higher than those in the normal medium group(P<0.05),while they were significantly lower than those in the 0%,5%,10%,25% and 30% drug-containing serum groups(P<0.05).It was found that the HIF-1α and VEGF-A gene expression in 15% drug-containing serum group was significantly lower than those of the 0%,5% and 10% drug-containing serum groups(P<0.05),while there was no significant difference between the 15% drug-containing serum group and the 20%,25% and 30% drug-containing serum groups(P>0.05).Conclusion:Bugu granules inhibit IL-1β-induced chondrocyte apoptosis by activating the HIF-1α/VEGF-A signaling pathway,and the concentration of drug-containing serum has effect on the activity of chondrocytes.

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备注/Memo

备注/Memo:
基金项目:福建省自然科学基金资助项目(2020J011191)
△通信作者 E-mail:18065142418@163.com
更新日期/Last Update: 2023-04-10