富血小板血浆对白细胞介素-1β诱导大鼠髓核细胞凋亡的影响

doi:10.3760/cma.j.cn441417-20241208-04014

国际医药卫生导报 ›› 2025, Vol. 31 ›› Issue (4): 591-596.DOI: 10.3760/cma.j.cn441417-20241208-04014

富血小板血浆对白细胞介素-1β诱导大鼠髓核细胞凋亡的影响

张国腾^1,2,4 王毅^1,2,4 周沛^1,2,4 邱世诚³ 周剑鹏^1,2,4

¹广州中医药大学附属广州中医医院骨科，广州 510130；²广州医科大学附属中医医院骨科，广州 510130；³江门市五邑中医院关节骨科，江门 529000；⁴广州医科大学中西医结合研究所，广州 510180

收稿日期:2024-12-08 出版日期:2025-02-15 发布日期:2025-02-24
通讯作者: 周剑鹏，Email：zjp800709@163.com
基金资助:
广东省医学科学技术研究基金（A2023282）；广州市科技计划（2024A03J0788）

Effect of platelet-rich plasma on IL-1β-induced apoptosis of rat nucleus pulposus cells

Zhang Guoteng^1,2,4, Wang Yi^1,2,4, Zhou Pei^1,2,4, Qiu Shicheng³, Zhou Jianpeng^1,2,4

¹Department of Orthopedics, The Affiliated Guangzhou Hospital of TCM of Guangzhou University of Chinese Medicine, Guangzhou 510130, China; ²Department of Orthopedics, The Affiliated Traditional Chinese Medicine Hospital, Guangzhou Medical University, Guangzhou 510130, China; ³Department of Arthrosis, Jiangmen Wuyi Hospital of Chinese Medicine, Jiangmen 529000, China; ⁴Institute of Integration of Traditional and Western Medicine of Guangzhou Medical University, Guangzhou 510180, China

Received:2024-12-08 Online:2025-02-15 Published:2025-02-24
Contact: Zhou Jianpeng, Email: zjp800709@163.com
Supported by:
Medical Science and Technology Research Foundation in Guangdong Province (A2023282); Guangzhou Science and Technology Plan (2024A03J0788)

摘要/Abstract

摘要：

目的　探讨富血小板血浆（PRP）对白细胞介素-1β（IL-1β）诱导大鼠髓核细胞凋亡的影响。方法　研究时间为2023年7月至2024年10月。采取SD大鼠血液，使用两步离心法提取PRP，用于后续实验。常规培养大鼠髓核细胞。采用不同浓度PRP对髓核细胞进行不同时间点的处理，用细胞活力检测试剂盒检测髓核细胞的活性。通过IL-1β诱导建立髓核细胞退变模型，随机分为3组：对照组（未经IL-1β和PRP处理）、IL-1β组（加入10 μg/L IL-1β溶液）和PRP组（加入10 μg/L IL-1β溶液和PRP）。干预72 h后，Western blot检测髓核组织中Bcl-2家族相关x蛋白（Bax）、B细胞淋巴瘤/白血病2基因（Bcl-2）以及半胱氨酸天冬氨酸蛋白酶-3蛋白（Caspase-3）表达水平；实时荧光定量聚合酶链式反应（qRT-PCR）检测Bax、Bcl-2以及Caspase-3 mRNA的表达。采用单因素方差分析及LSD-t检验进行数据分析。结果　Western blot检测结果显示，IL-1β组的Bax和Caspase-3蛋白表达量、Bax/Bcl-2均高于对照组（2.43±0.22比1.04±0.07、1.96±0.09比1.06±0.12、3.76±0.34比1.00±0.10），Bcl-2蛋白表达量低于对照组（0.65±0.05比1.05±0.17），差异均有统计学意义（均P<0.05）。PRP组的Bax和Caspase-3蛋白表达、Bax/Bcl-2低于IL-1β组，Bcl-2蛋白表达量高于IL-1β组，差异均有统计学意义（均P<0.05）；qRT-PCR检测结果显示，IL-1β组的Bax和Caspase-3 mRNA表达量均高于对照组（1.15±0.12比1.00±0.03、1.22±0.09比1.00±0.11），Bcl-2 mRNA表达量低于对照组（0.51±0.05比1.00±0.04），差异均有统计学意义（均P<0.05）。PRP组的Bax和Caspase-3 mRNA表达量低于IL-1β组，Bcl-2 mRNA表达量高于IL-1β组，差异均有统计学意义（均P<0.05）。结论　PRP可能抑制IL-1β诱导的髓核细胞凋亡和炎症反应，增强细胞活性，有效延缓椎间盘退变疾病进展。

关键词:

富血小板血浆, 白细胞介素-1β, 炎症反应, 细胞凋亡, 髓核细胞, 椎间盘退变, 动物实验

Abstract:

Objective　To observe the effect of platelet-rich plasma (PRP) on the apoptosis of rat nucleus pulposus cells induced by interleukin-1β (IL-1β). Methods　This study was from July 2023 to October 2024. The PRP was extracted from the SD rats' blood by the two-step centrifugation method, and was used for the subsequent experiments. The rat nucleus pulposus cells were routinely cultured in vitro. The nucleus pulposus cells were treated by different concentrations of PRP for different periods of time, and the cell viability test kit was used to detect the nucleus pulposus cells' activity. The degeneration model of the nucleus pulposus cells was established by IL-1β induction. The cells were randomly divided into a control group (without the treatment of IL-1β and PRP), an IL-1β group (adding 10 μg/L lL-1β solution), and a PRP group (adding 10 μg/L IL-1β solution and PRP). After 72 hours' intervention, the expression levels of Bcl-2 associated X protein (Bax), B-cell lymphoma/leukemia 2 (Bcl-2), and cysteine aspartic protease-3 (Caspase-3) were detected by the Western blotting. The expressions of Bax, Bcl-2, and Caspase-3 mRNA were detected by the quantitative real-time polymerase chain reaction (qRT-PCR). One-way analysis of variance and LSD-t test were used to analyze the data. Results　The western blotting test results show that the expressions of Bax and Caspase-3 protein and the ratio of Bax/Bcl-2 in the IL-1β group were higher than those in the control group (2.43±0.22 vs. 1.04±0.07, 1.96±0.09 vs. 1.06±0.12, and 3.76±0.34 vs. 1.00±0.10), while the expression of Bcl-2 protein was lower (0.65±0.05 vs. 1.05±0.17), with statistical differences (all P<0.05). The expressions of Bax and Caspase-3 proteins and the ratio of Bax/Bcl-2 in the PRP group were higher than those in the IL-1β group, while the expression of Bcl-2 protein was lower, with statistical differences (all P<0.05). The results of qRT-PCR showed that the expressions of Bax and Caspase-3 mRNA in the IL-1β group were higher than those in the control group (1.15±0.12 vs. 1.00±0.03 and 1.22±0.09 vs. 1.00±0.11), while the expression of Bcl-2 mRNA was lower (0.51±0.05 vs. 1.00±0.04), with statistical differences (all P<0.05). The expressions of Bax and Caspase-3 mRNA in the PRP group were lower than those in the IL-1β group, while the expression of Bcl-2 mRNA were higher, with statistical differences (all P<0.05). Conclusions　PRP can inhibit the apoptosis and inflammatory response of nucleus pulposus cells induced by IL-1β, enhance cell activity, and effectively delay the progression of intervertebral disc degeneration.

Key words:

Platelet-rich plasma, Interleukin-1β, Inflammatory response, Apoptosis, Nucleus pulposus cells, Intervertebral disc degeneration, Animal experiment

张国腾王毅周沛邱世诚周剑鹏.

富血小板血浆对白细胞介素-1β诱导大鼠髓核细胞凋亡的影响 [J]. 国际医药卫生导报, 2025, 31(4): 591-596.

Zhang Guoteng, Wang Yi, Zhou Pei, Qiu Shicheng, Zhou Jianpeng.

Effect of platelet-rich plasma on IL-1β-induced apoptosis of rat nucleus pulposus cells [J]. International Medicine and Health Guidance News, 2025, 31(4): 591-596.

参考文献

[1] Cao G, Yang S, Cao J, et al. The role of oxidative stress in intervertebral disc degeneration[J]. Oxid Med Cell Longev, 2022,2022:2166817. DOI: 10.1155/2022/2166817.
[2] Liu W, Jin S, Huang M, et al. Duhuo jisheng decoction suppresses matrix degradation and apoptosis in human nucleus pulposus cells and ameliorates disc degeneration in a rat model[J]. J Ethnopharmacol, 2020,250:112494. DOI: 10.1016/j.jep.2019.112494.
[3] Tian Y, Chu X, Huang Q, et al. Astragaloside IV attenuates IL-1β-induced intervertebral disc degeneration through inhibition of the NF-κB pathway[J]. J Orthop Surg Res, 2022,17(1):545. DOI: 10.1186/s13018-022-03438-1.
[4] Wang Y, Che M, Xin J, et al. The role of IL-1β and TNF-α in intervertebral disc degeneration[J]. Biomed Pharmacother, 2020,131:110660. DOI: 10.1016/j.biopha. 2020.110660.
[5] Akeda K, Ohishi K, Masuda K, et al. Intradiscal injection of autologous platelet-rich plasma releasate to treat discogenic low back pain: a preliminary clinical trial[J]. Asian Spine J, 2017,11(3):380-389. DOI: 10.4184/asj.2017.11.3.380.
[6] Levi D, Horn S, Tyszko S, et al. Intradiscal platelet-rich plasma injection for chronic discogenic low back pain: preliminary results from a prospective trial[J]. Pain Med, 2016,17(6):1010-22. DOI: 10.1093/pm/pnv053.
[7] 袁晨曦.富血小板血浆通过TGF-β1延缓椎间盘退变的初步研究[D].苏州:苏州大学,2014. DOI:10.7666/d.D504852.
[8] Chen WH, Lo WC, Lee JJ, et al. Tissue-engineered intervertebral disc and chondrogenesis using human nucleus pulposus regulated through TGF-beta1 in platelet-rich plasma[J]. J Cell Physiol, 2006,209(3):744-754. DOI: 10.1002/jcp.20765.
[9] Wang HQ, Yu XD, Liu ZH, et al. Deregulated miR-155 promotes Fas-mediated apoptosis in human intervertebral disc degeneration by targeting FADD and caspase-3[J]. J Pathol, 2011,225(2):232-242. DOI: 10.1002/path.2931.
[10] Boise LH, Gottschalk AR, Quintáns J, et al. Bcl-2 and Bcl-2-related proteins in apoptosis regulation[J]. Curr Top Microbiol Immunol, 1995,200:107-121. DOI: 10.1007/978-3-642-79437-7_8.
[11] Youle RJ, Strasser A. The BCL-2 protein family: opposing activities that mediate cell death[J]. Nat Rev Mol Cell Biol, 2008,9(1):47-59. DOI: 10.1038/nrm2308.
[12] 闫菁,胡江杉,胡霞,等. 针刀治疗对退变兔腰椎间盘组织中凋亡调控基因Bcl-2和Bax表达的影响[J]. 湖北中医药大学学报,2023,25(1):5-9. DOI:10.3969/j.issn.1008- 987x. 2023.01.01.
[13] 刘大锐,李报春,李怀东. 细胞凋亡核心基因Caspase家族的研究进展[J]. 中国医药导刊,2020,22(11):800-805. DOI:10.3969/j.issn.1009-0959.2020.11.014.
[14] Porter AG, Jänicke RU. Emerging roles of caspase-3 in apoptosis[J]. Cell Death Differ, 1999,6(2):99-104. DOI: 10.1038/sj.cdd.4400476.
[15] Rodrigues LMR, Oliveira LZ, Silva MBRD, et al. Inflammatory biomarkers in sera of patients with intervertebral disc degeneration[J]. Einstein (Sao Paulo), 2019,17(4):eAO4637. DOI: 10.31744/einstein_journal/2019AO4637.
[16] 李玲慧,王尚全,孙凯,等. 杜仲苷调控核糖体生成抑制髓核细胞损伤和细胞外基质降解的机制研究[J]. 中国中药杂志,2024,49(21):5713-5720. DOI:10.19540/j.cnki.cjcmm. 20240802.401.
[17] 李硕夫,刘宏哲,张嘉麟,等. 牛膝总皂苷对IL-1β诱导髓核细胞的凋亡及炎性损伤的影响[J]. 湖南中医药大学学报,2023,43(9):1591-1597. DOI:10.3969/j.issn.1674- 070X. 2023.09.007.
[18] Collins T, Alexander D, Barkatali B. Platelet-rich plasma: a narrative review[J]. EFORT Open Rev,2021,6(4):225-235. DOI: 10.1302/2058-5241.6.200017.
[19] 孙力,石辉. 富血小板血浆治疗膝关节炎的研究进展[J]. 国际医药卫生导报,2024,30(24):4106-4109. DOI:10.3760/cma.j.issn.1007-1245.2024.24.009.
[20] Alves R, Grimalt R. A review of platelet-rich plasma: history, biology, mechanism of action, and classification[J]. Skin Appendage Disord, 2018,4(1):18-24. DOI: 10.1159/000477353.
[21] Navani A, Manchikanti L, Albers SL, et al. Responsible, safe, and effective use of biologics in the management of low back pain: American Society of Interventional Pain Physicians (ASIPP) Guidelines[J]. Pain Physician, 2019,22(1S):S1-S74.
[22] Machado ES, Soares FP, Vianna de Abreu E, et al. Systematic review of platelet-rich plasma for low back pain[J]. Biomedicines, 2023,11(9):2404. DOI: 10.3390/biomedicines11092404.
[23] Navani A, Ambach M, Calodney A, et al. The safety and effectiveness of orthobiologic injections for discogenic chronic low back pain: a multicenter prospective, crossover, randomized controlled trial with 12 months follow-up[J]. Pain Physician, 2024,27(1):E65-E77.
[24] Nagae M, Ikeda T, Mikami Y, et al. Intervertebral disc regeneration using platelet-rich plasma and biodegradable gelatin hydrogel microspheres[J]. Tissue Eng, 2007,13(1):147-158. DOI: 10.1089/ten.2006.0042.

富血小板血浆对白细胞介素-1β诱导大鼠髓核细胞凋亡的影响

Effect of platelet-rich plasma on IL-1β-induced apoptosis of rat nucleus pulposus cells

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