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新疆维吾尔自治区人民医院骨科中心关节运动病区,新疆 乌鲁木齐 830001
王志刚,E-mail: 13899809527@163.com
收稿日期:2024-06-18,
纸质出版日期:2025-09-25
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孟晓源,安梦宇,王志刚等.α-倒捻子素改善大鼠软骨细胞的凋亡和炎症反应的作用机制[J].局解手术学杂志,2025,34(09):759-764.
MENG Xiao-yuan,AN Meng-yu,WANG Zhi-gang,et al.Mechanism of alpha-mangostin improving apoptosis and inflammatory response in rat chondrocytes[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(09):759-764.
孟晓源,安梦宇,王志刚等.α-倒捻子素改善大鼠软骨细胞的凋亡和炎症反应的作用机制[J].局解手术学杂志,2025,34(09):759-764. DOI: 10.11659/jjssx.06E024072.
MENG Xiao-yuan,AN Meng-yu,WANG Zhi-gang,et al.Mechanism of alpha-mangostin improving apoptosis and inflammatory response in rat chondrocytes[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(09):759-764. DOI: 10.11659/jjssx.06E024072.
目的
2
探索α-倒捻子素(α-MG)对脂多糖(LPS)诱导的体外大鼠软骨细胞骨关节炎(OA)和巨噬细胞M1型极化的影响。
方法
2
用LPS诱导大鼠软骨细胞体外OA模型。将大鼠软骨细胞和巨噬细胞分为对照组(正常培养)、LPS组(使用1 μg/mL LPS刺激24 h)、LPS+α-MG组(使用40 μmol/L α-MG联合1 μg/mL LPS刺激细胞24 h)、LPS+α-MG+si-NC组(使用70 μg/mL si-NC转染并联合40 μmol/L α-MG及1 μg/mL LPS共同刺激细胞24 h)、LPS+α-MG+si-BDNF组(使用70 μg/mL si-BDNF转染并联合40 μmol/L α-MG及1 μg/mL LPS共同刺激细胞24 h)。CCK-8法检测各组软骨细胞和巨噬细胞增殖活性,流式细胞术检测软骨细胞的凋亡和巨噬细胞极化。ELISA法检测各组软骨细胞中白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)-α水平。Western blot检测软骨细胞和巨噬细胞中BDNF、PI3K、Akt表达。
结果
2
在软骨细胞中,与对照组比较,LPS组的细胞增殖率降低,细胞凋亡率增加,BDNF、PI3K、Akt蛋白表达下调,而IL-1β、IL-6、TNF-α水平上调(
P
<
0.05);与LPS组比较,LPS+α-MG组细胞增殖率增加,细胞凋亡率降低,BDNF、PI3K、Akt蛋白表达上调,而IL-1β、IL-6、TNF-α水平下调(
P
<
0.05);与LPS+α-MG+si-NC组比较,LPS+α-MG+si-BDNF组细胞增殖率降低,细胞凋亡率增加,BDNF、PI3K、Akt蛋白表达下调,而IL-1β、IL-6、TNF-α水平均升高(
P
<
0.05)。在巨噬细胞中,与对照组比较,LPS组细胞增殖率降低,M1型细胞比例增加,而BDNF、PI3K、Akt蛋白表达下调(
P
<
0.05);与LPS组比较,LPS+α-MG组细胞增殖率增加,M1型细胞比例降低,而BDNF、PI3K、Akt蛋白表达上调(
P
<
0.05);与LPS+α-MG+si-NC组比较,LPS+α-MG+si-BDNF组细胞增殖率降低,M1型细胞比例增加,而BDNF、PI3K、Akt蛋白表达水平下调(
P
<
0.05)。
结论
2
α-MG通过激活BDNF/PI3K/Akt信号通路抑制巨噬细胞M1型极化,改善大鼠软骨细胞的凋亡和炎症。
Objective
2
To explore the effects of alpha-mangostin (α-MG) on the in vitro osteoarthritis (OA) in rat chondrocytes induced by lipopolysaccharide (LPS) and the M1 polarization of macrophages.
Methods
2
An in vitro OA model of rat chondrocytes was induced by LPS. Both rat chondrocytes and macrophages were divided into the control group (normal culture),LPS group (stimulated with 1 μg/mL LPS for 24 hours),LPS+α-MG group (stimulated with 40 μmol/L α-MG combined with 1 μg/mL LPS for 24 hours),LPS+α-MG si-NC group (transfected with 70 μg/mL si-NC and stimulated with 40 μmol/L α-MG and 1 μg/mL LPS for 24 hours),and LPS+α-MG+si-BDNF group (transfected with 70 μg/mL si-BDNF and stimulated with 40 μmol/L α-MG and 1 μg/mL LPS for 24 hours). CCK-8 assay was used to detect the proliferative viability of chondrocytes and macrophages in each group,and flow cytometry was used to detect the apoptosis of chondrocytes and the polarization of macrophages. ELISA was used to detect the levels of interleukin(IL)-1β,IL-6,and tumor necrosis factor (TNF)-α of chondrocytes in each group. Western blot was used to detect the expression of BDNF,PI3K,and Akt in chondrocytes and macrophages.
Results
2
In chondrocytes,compared with the control group,the LPS group had decreased cell proliferation rate,increased cell apoptosis rate,down-regulated expression of BDNF,PI3K and Akt proteins,and up-regulated levels of IL-1β,IL-6 and TNF-α (
P
<
0.05); compared with the LPS group,the LPS+α-MG group showed increased cell proliferation rate,decreased cell apoptosis rate,up-regulated expression of BDNF,PI3K and Akt proteins,and down-regulated levels of IL-1β,IL-6 and TNF-α (
P
<
0.05); compared with the LPS+α-MG+si-NC group,the LPS+α-MG si-BDNF group demonstrated decreased cell proliferation rate,increased cell apoptosis rate,down-regulated expression of BDNF,PI3K and Akt proteins,and increased levels of IL-1β,IL-6 and TNF-α (
P
<
0.05). In macrophages,compared with the control group,the LPS group had decreased cell proliferation rate,increased proportion of M1 cells,and down-regulated expression of BDNF,PI3K and Akt proteins (
P
<
0.05); compared with the LPS group,the LPS+α-MG group showed increased cell proliferation rate,decreased proportion of M1 c
ells,and up-regulated expression of BDNF,PI3K and Akt proteins (
P
<
0.05); compared with the LPS+α-MG+si-NC group,the LPS+α-MG+si-BDNF group showed decreased cell proliferation rate,increased proportion of M1 cells,and down-regulated expression of BDNF,PI3K and Akt proteins (
P
<
0.05).
Conclusion
2
α-MG inhibits the M1 polarization of macrophages to improve the apoptosis and inflammation of rat chondrocytes by activating the BDNF/PI3K/Akt signaling pathway.
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