| 二氢杨梅素通过ROS/p38-MAPK信号通路对H2O2诱导骨关节炎软骨细胞增生、凋亡的影响 |
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Received:September 18, 2023
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| 作者 | Author | 单位 | Unit | E-Mail |
| 程瑛 |
CHENG Ying |
武汉市中医医院骨伤科, 湖北 武汉 430000 |
Department of Orthopaedics and Traumatology, Wuhan Hospital of Traditional Chinese Medicine, Wuhan 430000, Hubei, China |
elzklm@163.com |
| 陈慧娟 |
CHEN Hui-juan |
武汉市中医医院骨伤科, 湖北 武汉 430000 |
Department of Orthopaedics and Traumatology, Wuhan Hospital of Traditional Chinese Medicine, Wuhan 430000, Hubei, China |
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| 杨婷 |
YANG Ting |
武汉市中医医院骨伤科, 湖北 武汉 430000 |
Department of Orthopaedics and Traumatology, Wuhan Hospital of Traditional Chinese Medicine, Wuhan 430000, Hubei, China |
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| 期刊信息：《中国骨伤》2025年38卷，第4期，第396-402页 |
| DOI：10.12200/j.issn.1003-0034.20230237 |
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目的: 分析二氢杨梅素通过活性氧(reactive oxygen,ROS)/p38丝裂原活化蛋白激酶(p38 mitogen-acti-vated protein kinase,p38-MAPK)通路对过氧化氢(hydrogen peroxide,H2O2)诱导的骨关节炎软骨细胞增生、凋亡的影响。
方法: 取5只C57BL/6J小鼠,麻醉后颈椎脱臼法处死,提取并培养软骨细胞,传代后培养细胞,将对数生长期软骨细胞分为对照组、H2O2组(0.8 μmol·L-1的H2O2)、二氢杨梅素低浓度组(0.8 μmol·L-1的H2O2+20 μmol·L-1二氢杨梅素)、二氢杨梅素高浓度组(0.8 μmol·L-1的H2O2+80 μmol·L-1二氢杨梅素)、ROS抑制剂N-乙酰半胱氨酸(N-acetyl-cysteine,NAC)组(0.8 μmol·L-1的H2O2+5 mmol·L-1 NAC),噻唑蓝(methyl thiazolyl tetrazolium,MTT)法测定软骨细胞活性;Hoechst 33342法测定软骨细胞凋亡率;2,7-二氯荧光素二乙酸酯(2,7-dichlorofluorescein diacetate,DCFH-DA)荧光探针测定软骨细胞内ROS水平;qRT-PCR法测定Ⅱ型胶原α1(typeⅡcollagen α1,Col2α1) mRNA水平;Western blot法测定Col2α1、p-p38-MAPK/p38-MAPK、B细胞淋巴瘤基因-2(B cell lymphoma gene-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)蛋白表达。
结果: 软骨细胞呈旋涡状纤维团,COL2α表达阳性;与对照组相比,H2O2组软骨细胞活力、凋亡率、ROS荧光强度、p-p38-MAPK/p38-MAPK、Bax蛋白表达升高,Col2α1 mRNA水平、Col2α1、Bcl-2蛋白表达降低,差异有统计学意义(P<0.05);与H2O2组相比,二氢杨梅素低浓度组、二氢杨梅素高浓度组、NAC组软骨细胞活力、凋亡率、ROS荧光强度、p-p38-MAPK/p38-MAPK、Bax蛋白表达降低,Col2α1 mRNA水平、Col2α1、Bcl-2蛋白表达升高,差异有统计学意义(P<0.05)。
结论: 二氢杨梅素可能通过抑制ROS/p38-MAPK通路抑制软骨细胞凋亡、炎性反应、氧化应激,二氢杨梅素可能是治疗骨关节炎的潜在药物。 |
| [关键词]:二氢杨梅素 骨关节炎 软骨细胞增生 凋亡 ROS/p38-MAPK信号通路 |
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| Influences of dihydromyricetin on proliferation and apoptosis of chondrocytes in osteoarthritis induced by H2O2 through ROS/p38-MAPK signal pathway |
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Abstract:
Objective To analyze the influences of dihydromyricetin on the proliferation and apoptosis of chondrocytes in osteoarthritis induced by hydrogen peroxide (H2O2) through reactive oxygen species (ROS)/p38 mitogen activated protein kinase (p38-MAPK) pathway.
Methods Five C57BL/6J mice were euthanized by cervical dislocation after anesthesia. Chondrocytes were extracted and cultured.After passage,the chondrocytes were divided into control group,H2O2 group (0.8 μmol·L-1 H2O2),dihydromyricetin low concentration group (0.8 μmol·L-1 H2O2+20 μmol·L-1 dihydromyricetin),dihydromyricetin high concentration group (0.8 μmol·L-1 H2O2+80 μmol·L-1 dihydromyricetin),and ROS inhibitor N-acetylcysteine (NAC) group (0.8 μmol·L-1 H2O2+5 mmol·L-1 NAC). The activity of chondrocytes was measured by methyl thiazolyl tetrazolium (MTT) assay. The apoptosis rate of chondrocytes was measured by Hoechst 33342 method. The level of ROS in chondrocytes was measured by 2,7-dichlorofluorescein diacetate (DCFH-DA) fluorescence probe.The level of Type II collagen α1 (Col2α1) mRNA was measured by qRT-PCR.And the expression of Col2α1,p-p38-MAPK/p38-MAPK,B cell lymphoma gene-2 (Bcl-2) and Bcl-2 associated X protein (Bax) proteins was detected by Western blot.
Results The chondrocytes showed swirling fibrous mass,and the expression of COL2α was positive. Compared with the control group,the chondrocyte viability,apoptosis rate,ROS fluorescence intensity,p-p38-MAPK/p38-MAPK,and the expression of Bax protein in H2O22 group increased,the level of Col2α1 mRNA,and the expression of Col2α1 and Bcl-2 proteins decreased (P<0.05). Compared with H2O2 group,the chondrocyte viability,apoptosis rate,ROS fluorescence intensity,p-p38-MAPK/p38-MAPK,and the expression of Bax pro-tein in dihydromyricetin low concentration group,dihydromyricetin high concentration group,and NAC group decreased,the level of Col2α1 mRNA,and the expression of Col2α1 and Bcl-2 proteins increased (P<0.05).
Conclusion Dihydromyricetin may inhibit chondrocyte apoptosis,inflammatory reaction and oxidative stress by inhibiting ROS/p38-MAPK pathway. Dihydromyricetin may be a potential drug for treating osteoarthritis. |
| KEYWORDS:Dihydromyricetin Osteoarthritis Chondrocyte proliferation Apoptosis ROS/p38 -MAPK signal pathway |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 程瑛,陈慧娟,杨婷.二氢杨梅素通过ROS/p38-MAPK信号通路对H2O2诱导骨关节炎软骨细胞增生、凋亡的影响[J].中国骨伤,2025,38(4):396~402 |
| 英文格式: | CHENG Ying,CHEN Hui-juan,YANG Ting.Influences of dihydromyricetin on proliferation and apoptosis of chondrocytes in osteoarthritis induced by H2O2 through ROS/p38-MAPK signal pathway[J].zhongguo gu shang / China J Orthop Trauma ,2025,38(4):396~402 |
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