Objective  To investigate the effect of fumigation decoction on the expression of aggrecan (AGG) and type II collagen (COL2A1) in an intervertebral disc degeneration (IDD) model,  and to gain an in-depth understanding of the cellular and molecular mechanisms underlying traditional Chinese medicine fumigation therapy.

Methods  An IDD model was established using Sprague-Dawley rats by inducing disc injury via needle puncture. The rats were divided into three groups: normal control (CON), model (PUN), and fumigation (CHF). Only the CHF group received treatment with fumigation decoction for 4 weeks. Imaging analysis was conducted to assess the relative height of intervertebral discs, while Masson's trichrome staining was used to observe structural changes in the discs. The expression of NLRP3 inflammasome was detected by immunohistochemistry. Quantitative RT-PCR and Western blotting were employed to evaluate the expression levels of AGG and COL2A1 genes and proteins.

Results  A total of 12 rats were randomly divided into 3 groups, with 4 rats in each group. Compared with the CON group, disc height was significantly lost in the PUN group (P<0.05), and disc height could be partially restored in the CHF group (P<0.05). In the PUN group, the expression levels of AGG mRNA and COL2A1 mRNA and protein in the nucleus pulposus tissue were significantly lower than those in the CON group (P<0.05). Compared with the PUN group, the CHF group showed significantly increased expression levels of AGG mRNA and COL2A1 mRNA and protein in the nucleus pulposus tissue (P<0.05). Histological analysis revealed that, compared with the PUN group, the CHF group exhibited partial restoration of the gel-like properties of the nucleus pulposus and an increase in proteoglycan content. Additionally, NLRP3 expression in the nucleus pulposus tissue of the CHF group was markedly reduced.

Conclusion  Fumigation decoction treatment can enhance the expression of AGG and COL2A1 and decrease the expression of NLRP3 in IDD models, which may be one of its mechanisms for treating lumbar degenerative diseases. This research provides new insights into the treatment of IDD.

1.Ravindra VM, Senglaub SS, Rattani A, et al. Degenerative lumbar spine disease: estimating global incidence and worldwide volume[J]. Global Spine J, 2018, 8(8): 784-794. DOI: 10.1177/2192568218770769.

2.Roughley PJ. Biology of intervertebral disc aging and degeneration: involvement of the extracellular matrix[J]. Spine (Phila Pa 1976), 2004, 29(23): 2691-2699. DOI: 10.1097/01.brs.0000146101.53784.b1.

3.吴晓东, 袁文. 腰椎间盘突出症治疗方法的进展和并发症的防范[J]. 中国骨伤, 2024, 37(3): 219-221. [Wu XD, Yuan W. Progress on treatment and prevention of complications of lumbar disc herniation[J]. China Journal of Orthopaedics and Traumatology, 2024, 37(3): 219-221.] DOI: 10.12200/j.issn.1003-0034.20240123.

4.韩涛, 于杰, 金哲峰, 等. 中药干预椎间盘退变治疗腰椎间盘突出症作用机制研究进展[J]. 世界中医药, 2023, 18(21): 3042-3046. [Han T, Yu J, Jin ZF, et al. Research progress in the mechanisms of Chinese medicines in the treatment of lumbar disc herniation due to intervertebral disc degeneration[J]. World Chinese Medicine, 2023, 18(21): 3042-3046.] DOI: 10.3969/j.issn.1673- 7202.2023.21.007.

5.李艳, 陆华岭, 吴耀持. 基于“肝主筋”理论治疗腰椎间盘突出症思路探析[J]. 江苏中医药, 2014, (11): 6-7. [Li Y,Lu HL, Wu YC. Exploration of therapeutic strategies for lumbar disc herniation based on the theory of "Liver Governing Sinews"[J].Jiangsu Journal of Traditional Chinese Medicine, 2014, (11): 6-7.] https://d.wanfangdata.com.cn/periodical/jszy201411003.

6.Zhu L, Yu C, Zhang X, et al. The treatment of intervertebral disc degeneration using traditional Chinese medicine[J]. J Ethnopharmacol, 2020, 263: 113117. DOI: 10.1016/j.jep.2020. 113117.

7.Hua W, Li S, Luo R, et al. Icariin protects human nucleus pulposus cells from hydrogen peroxide-induced mitochondria-mediated apoptosis by activating nuclear factor erythroid 2-related factor 2[J]. Biochim Biophys Acta Mol Basis Dis, 2020, 1866(1): 165575. DOI: 10.1016/j.bbadis.2019.165575.

8.李兆勇, 张晨阳, 郭彦涛, 等. 基于数据挖掘的中医医案古籍腰痛症用药规律研究[J]. 湖南中医药大学学报, 2020, 40(10): 1280-1282. [Li ZY, Zhang CY, Guo YT, et al. Research on prescriptions and herbs for lumbago in ancient books of traditional Chinese medical records based on data mining[J]. Journal of Hunan University of Chinese Medicine, 2020, 40(10): 1280-1282.] DOI: 10.3969/j.issn.1674-070X.2020.10.021.

9.吴国林, 何元诚. 中医药治疗腰椎间盘突出症的治疗进展[J]. 中国中医骨伤科杂志, 2010, 18(5): 70-72. https://d.wanfangdata.com.cn/periodical/zgzygsk201005033.

10.陈铎葆, 徐冰, 李兵, 等. 寻骨风对抗炎、镇痛作用的研究 [J]. 基层中药杂志, 2001, 15(1): 9-10. [Chen DB, Xu B, Li B, et al. Study on the anti-inflammation and analgesia of Aristolochia mollissima Hance[J]. Primary Journal Of Chinese Materia Medica, 2001, 15(1): 9-10.] DOI: 10.3969/j.issn.1673-6427.2001.01.004.

11.杨秀娟, 邓毅, 杨志军, 等. 当归不同药用部位对急性血瘀模型大鼠的活血作用[J]. 中国临床药理学杂志, 2018, 34(18): 2187-2190. [Yang XJ, Deng Y, Yang ZJ, et al. Effect of different medicinal part in Angelica sinensis on the promoting blood in acute blood stasis rats[J]. The Chinese Journal of Clinical Pharmacology, 2018, 34(18): 2187-2190.] DOI: 10.13699/j.cnki.1001-6821.2018.18.016.

12.蒋丽, 郭瑞齐, 管仁伟, 等. 凤仙透骨草HPLC指纹图谱及其与抗炎活性的谱效关系[J]. 山东中医药大学学报, 2022, 46(6): 747-757. [Jiang L, Guo RQ, Guan RW, et al. HPLC fingerprint of Fengxiantougucao(impatiens balsamina L.) and spectral-effect relationship with anti-inflammatory activity[J]. Journal of Shandong University of Traditional Chinese Medicine, 2022, 46(6): 747-757.] DOI: 10.16294/j.cnki.1007-659x.2022.06.014.

13.李照辉, 熊圣仁, 郑竑, 等. 海桐皮汤熏洗治疗跟痛症130例临床观察[J]. 风湿病与关节炎, 2021, 10(11): 10-12, 38. [Li ZH, Xiong SR, Zheng H, et al. Clinical observation on 130 cases of painful heel syndrome treated by fumigation and washing with haitongpi tang[J]. Rheumatism and Arthritis, 2021, 10(11): 10-12, 38.] DOI: 10.3969/j.issn.2095-4174.2021.11.003.

14.赵睿晞, 王人彦. 张玉柱教授治疗腰椎间盘突出症的用药规律分析[J]. 中医正骨, 2021, 33(8): 39-42. [Zhao RX, Wang RY. An analysis of clinical medication rules of Professor Zhang Yuzhu in treatment of lumbar disc herniation[J]. The Journal of Traditional Chinese Orthopedics and Traumatology, 2021, 33(8): 39-42.] DOI: 10.3969/j.issn.1001-6015.2021.08.008.

15.朱杰, 于立芹, 张华南, 等. 伸筋草经皮给药对小鼠抗炎镇痛作用及机制研究[J]. 中医药导报, 2021, 27(6): 40-44. [Zhu J, Yu LQ, Zhang HN, et al. Study on the anti-inflammatory and analgesic effects and mechanism of the transdermal administration of Shenjincao(lycopodii herba)on mice[J]. Guiding Journal of Traditional Chinese Medicine and Pharmacology, 2021, 27(6): 40-44.] DOI: 10.13862/j.cnki.cn43-1446/r.2021.06.017.

16.边甜甜, 司昕蕾, 牛江涛, 等. 花椒挥发油部位经清炒法炮制前后在小鼠体内的抗炎与镇痛作用[J]. 中国临床药理学杂志, 2019, 35(4): 369-371, 376. [Bian TT, Si XL, Niu JT, et al. Anti-inflammatory and analgesic effects of Volatile oil from Zanthoxylum bungeanum Maxim before and after processing by stir frying in mice[J]. The Chinese Journal of Clinical Pharmacology, 2019, 35(4): 369-371, 376.] DOI: 10.13699/j.cnki.1001-6821. 2019.04.015.

17.娄纯彪, 刘蓓, 刘茜, 等. 中药熏洗联合局部注射治疗跟痛症的临床研究[J]. 中国中医骨伤科杂志, 2019, 27(3): 20-22, 28. [Lou CB, Liu B, Liu Q, et al. The clinical research of Herbal fumigation and wash therapy combined with local injection in treating calcaneodynia[J]. Chinese Journal of Traditional Medical Traumatology & Orthopedics, 2019, 27(3): 20-22, 28.] https://d.wanfangdata.com.cn/periodical/zgzygsk201903005.

18.周稆, 梁玉婷, 邱辕鑫, 等. 中药熏蒸治疗腰椎间盘突出症的用药规律探析[J]. 湖南中医杂志, 2023, 39(4): 31-34, 73. [Zhou L, Liang YT, Qiu YX, et al. Medication rule of traditional Chinese medicine fumigation and steaming therapy in treatment of lumbar disc herniation[J]. Hunan Journal of Traditional Chinese Medicine, 2023, 39(4): 31-34, 73.] DOI: 10.16808/j.cnki.issn 1003-7705.2023.04.007.

19.Wei Q, Zhang X, Zhou C, et al. Roles of large aggregating proteoglycans in human intervertebral disc degeneration[J]. Connect Tissue Res, 2019, 60(3): 209-218. DOI: 10.1080/ 03008207.2018.1499731.

20.Wei Y, Tower RJ, Tian Z, et al. Spatial distribution of type II collagen gene expression in the mouse intervertebral disc[J]. JOR Spine, 2019, 2(4): e1070. DOI: 10.1002/jsp2.1070.

21.Chen F, Lei L, Chen S, et al. Serglycin secreted by late-stage nucleus pulposus cells is a biomarker of intervertebral disc degeneration[J]. Nat Commun, 2024, 15(1): 47. DOI: 10.1038/s41467-023-44313-9.

22.Hao Z, Tao C, Aimin WU. Effects of oxidative stress on mitochondrial functions and intervertebral disc cells[J]. Journal of Sichuan University (Medical Science Edition), 2024, 55(2): 249-255. DOI: 10.12182/20240360201.

23.徐浩伟, 房心月, 常圣杰, 等. 延缓椎间盘退变的抗衰老疗法研究进展[J]. 中国矫形外科杂志, 2024, 32(19): 1770-1775. [Xu HW, Fang XY, Chang SJ, et al. Research progress of anti-aging therapy to delay disc degeneration[J]. Orthopedic Journal of China, 2024, 32(19): 1770-1775.] DOI: 10.20184/j.cnki.Issn1005-8478.100462.

24.Chi S, Li S, Xu Z, et al. The involvement of DDX3X in compression-induced nucleus pulposus pyroptosis[J]. Biochem Biophys Res Commun, 2023, 655: 1-10. DOI: 10.1016/j.bbrc.2023.02.074.