| 卿海辉,张小舟,胡敏.蒲公英提取物调控程序化死亡基因 5对骨关节炎软骨细胞增殖、凋亡的影响及其机制研究[J].安徽医药,2021,25(9):1717-1722. |
| 蒲公英提取物调控程序化死亡基因 5对骨关节炎软骨细胞增殖、凋亡的影响及其机制研究 |
| Effect of dandelion extract on proliferation and apoptosis of chondrocyte in osteoarthritis by regulation of PDCD5 and its mechanism |
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| DOI：10.3969/j.issn.1009-6469.2021.09.005 |
| 中文关键词: 蒲公英 程序化死亡基因 5 骨关节炎 软骨细胞 增殖 凋亡 |
| 英文关键词: Taraxacum mongolicum Programmed death gene 5 Osteoarthritis Chondrocytes Proliferation Apoptosis |
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| 中文摘要: |
| 目的探究蒲公英提取物通过调控程序化死亡基因 5(PDCD5)的表达对骨关节炎软骨细胞增殖、凋亡的影响及其作用机制。方法选取 2018年 1月至 2019年 6月武汉科技大学附属孝感医院获得接受关节置换的 5例骨关节炎病人骨组织，从中分离、培养骨关节炎软骨细胞，蒲公英提取物高、中、低剂量组处理软骨细胞， MTT法检测细胞增殖，流式细胞仪检测细胞凋亡，蛋白质印迹法检测细胞中细胞周期蛋白 D1(cyclin D1)、细胞周期蛋白依赖性激酶抑制剂 1A(P21)、 B细胞淋巴瘤 -2(Bcl2)、 Bcl-2相关 X(Bax)和 PDCD5蛋白表达水平，实时荧光定量 PCR检测细胞 PDCD5 mRNA表达水平；过表达或者沉默 PDCD5基因，检测其对骨关节炎软骨细胞增殖、凋亡的影响。结果与对照组相比，蒲公英提取物 2.5 g/L、5.0 g/L、10.0 g/L组软骨细胞活性升高［48 h:(0.68±0.05)、(0.83±0.06)、(1.06±0.06)比( 0.56±0.03)； 72 h:(0.97±0.07)、(1.42±0.07)、(1.89±0.07)比( 0.79±0.05)］细胞凋亡率降低［(19.40±1.10)%、(12.31±0.59)%、(7.68±0.54)%比( 22.10±1.20)%］； P21、Bax表达水平降低， cyclin D1、 Bcl-2水平升高， PDCD5 mRNA和蛋白表达水平降低( P<0.05)。与 si-NC组比较， si-PDCD5组软骨细胞活性升高［48 h:表达，(0.86±0.05)比( 0.56±0.03)； 72 h:(1.33±0.07)比( 0.79±0.05)］细胞凋亡率降低［( 14.23±1.02)%比( 21.98±1.12)%］； P21、Bax表达水平降低， cyclin D1、Bcl-2表达水平升高( P<0.05)。与蒲公英，提取物 5 g/L+pcDNA3.1组比较，蒲公英提取物 5 g/L+pcDNAPDCD5组软骨细胞活性降低［48 h:(0.70±0.05)比( 0.86±0.06)； 72 h:(1.15±0.07)比( 1.51±0.07)］，细胞凋亡率升高［( 16.88±1.02)%比( 9.56±0.63)%］； P21、Bax表达水平升高， cyclin D1、Bcl-2表达水平降低( P<0.05)。结论蒲公英提取物可促进软骨细胞的增殖，抑制其凋亡，其可能通过调控 PDCD5基因的表达发挥对骨关节炎软骨细胞的增殖促进、凋亡抑制作用。 |
| 英文摘要: |
| Objective To investigate the effects of Taraxacum mongolicum extract on osteoarthritis chondrocyte proliferation andapoptosis by regulating the expression of programmed cell death 5 (PDCD5) and its mechanism.Methods The bone tissues of 5 patients with osteoarthritis, who underwent joint replacements in Xiaogan Hospital Affiliated to Wuhan University of Science and Technology from January 2018 to June 2019, were selected, and osteoarthritis chondrocytes were isolated, cultured, and treated with Taraxacummongolicum at high, medium and low doses. Methyl Thiazolyl Tetrazolium (MTT) method was used to detect cell proliferation, flow cytometry was used to detect cell apoptosis, Western blotting was used to detect cyclin D1, Cyclin-dependent kinase inhibitor 1A (P21), B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2 related X (Bax) and PDCD5 protein expression levels, and real-time fluorescent quantitative PCR was used to detect PDCD5 mRNA expression level. Detection was conducted of the effect of over-expression or silencing of PDCD5 gene on the proliferation and apoptosis of osteoarthritis chondrocytes.Results Compared with the control group, in Taraxacummongolicum extract 2.5 g/L, 5.0 g/L, 10.0 g/L group chondrocyte activity was increased [48 h: (0.68±0.05), (0.83±0.06), (1.06±0.06) vs. (0.56±0.03); 72 h: (0.97±0.07), (1.42±0.07), (1.89±0.07) vs. (0.79±0.05)], the cell apoptosis rate was decreased [(19.40±1.10)% , (12.31± 0.59)%, (7.68±0.54)% vs. (22.10±1.20)%]; P21, Bax expression levels were decreased, cyclin D1, Bcl-2 expression levels were increased, and PDCD5 mRNA and protein expression levels were decreased (P<0.05). Compared with the si-NC group, in the siPDCD5 group the activity of chondrocytes was increased [48 h: (0.86±0.05) vs. (0.56±0.03); 72 h: (1.33±0.07) vs. (0.79±0.05)], the cell apoptosis rate was decreased [(14.23±1.02)% vs. (21.98±1.12)%], the expression levels of P21 and Bax were decreased, and the expression levels of cyclin D1 and Bcl-2 were increased (P<0.05). Compared with the Taraxacum mongolicum extract 5 g/L+pcDNA3.1 group, in Taraxacum mongolicum extract 5 g/L+pcDNA-PDCD5 group chondrocyte activity was decreased [48 h: (0.70±0.05) vs. (0.86±0.06); 72 h: (1.15±0.07) vs. (1.51±0.07)], cell apoptosis rate was increased [(16.88±1.02)% vs. (9.56±0.63)%], P21 and Bax expression levels were increased, and cyclin D1 and Bcl-2 expression levels were decreased (P<0.05).Conclusions Taraxacum mongolicum extract canpromote the proliferation of chondrocytes and inhibit their apoptosis. It may play an important role in promoting the proliferation and inhibiting the apoptosis of osteoarthritis chondrocytes by regulating the expression of PDCD5 gene. |
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