Transcriptome sequencing-based study on the mechanism of action of Jintiange capsules (金天格胶囊) in regulating synovial mesenchymal stem cells exosomal miRNA and articular chondrocytes mRNA for the treatment of osteoarthritis

doi:10.19852/j.cnki.jtcm.20240927.004

Abstract

Abstract:

OBJECTIVE: To corroborate the efficacy of Jintiange capsules (JTGs) (金天格胶囊) in the treatment of osteoarthritis (OA) by exploring the potential mechanism of action of synovial mesenchymal stem cell exosomes (SMSC-Exos) and articular chondrocytes (ACs) through transcriptome sequencing (RNA-seq).

METHODS: Type II collagenase was used to induce OA in rats. The efficacy of JTGs was confirmed by macroscopic observation of articular cartilage, micro-CT observation, and safranin fast green staining. After SMSC-Exos and ACs were qualified, RNA-seq was used to screen differentially expressed miRNAs and mRNAs. The target genes of differentially expressed miRNAs in Synovial mesenchymal stem cells (SMSCs) were predicted based on the multiMiR R package. The co-differentially expressed genes of SMSC-Exos and ACs were obtained by venny 2.1.0. The miRNA-mRNA regulatory network was constructed by Cytoscape software. Based on the OmicShare platform, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis was performed on the mRNA regulated by key miRNAs. Expression trend analysis was performed for co-differentially expressed genes. Correlation analysis was performed on micro-CT efficacy indicators, co-differentially expressed genes mRNA and miRNA.

RESULTS: The efficacy of each administration group of JTGs was significant compared with the model group. SMSC-Exos and ACs were identified by their characteristics. The expression of rno-miR-23a-3p, rno-miR-342-3p, rno-miR-146b-5p, rno-miR-501-3p, rno-miR-214-3p was down-regulated in OA pathological state, and the expression of rno-miR-222-3p, rno-miR-30e-3p, rno-miR-676, and rno-miR-192-5p expression was up-regulated, and the expression of all these miRNAs was reversed after the intervention with JTGs containing serum. The co-differentially expressed genes were enriched in the interleukin 17 signaling pathway, tumor necrosis factor signaling pathway, transforming growth factor-β signaling pathway, etc. The expression trends of Ccl7, Akap12, Grem2, Egln3, Arhgdib, Ccl20, Mmp12, Pla2g2a, and Nr4a1 were significant. There was a correlation between micro-CT pharmacodynamic index, mRNA, and miRNA.

CONCLUSION: JTGs can improve the degeneration of joint cartilage and achieve the purpose of cartilage protection, which can be used for the treatment of OA. SMSCs-related miRNA expression profiles were significantly altered after the intervention with JTGs containing serum. The 9 co-differentially expressed genes may be the key targets for the efficacy of JTGs in the treatment of OA rats, which can be used for subsequent validation.

Key words: transcriptome sequencing technology, osteoarthritis, Jintiange capsules, synovial mesenchymal stem cells, articular chondrocytes

CHEN Zhongying, ZHANG Xue, ZHANG Xiaofei, ZOU Junbo, YUAN Puwei, SHI Yajun. Transcriptome sequencing-based study on the mechanism of action of Jintiange capsules (金天格胶囊) in regulating synovial mesenchymal stem cells exosomal miRNA and articular chondrocytes mRNA for the treatment of osteoarthritis[J]. Journal of Traditional Chinese Medicine, 2024, 44(6): 1153-1167.

References 45.

1.	Sellam J, Berenbaum F. The role of synovitis in pathophysiology and clinical symptoms of osteoarthritis. Nat rev rheumato 2010; 6: 625-35.
2.	Scanzello C, Goldring S. The role of synovitis in osteoarthritis pathogenesis. Bone 2012; 51: 249-57. DOI PMID
3.	Zhao Y, Li A, Ni L, Wang W, Shi X. The research progress of tiger bone and artificial tiger bone for the treatment of osteoporosis. Zhong Guo Gu Zhi Shu Song Za Zhi 2012; 18: 95-8.
4.	Sun J, Yang X, Hu Y. Efficacy of Jintiange Capsules in the treatment of osteoporosis: a network Meta-analysis. Orthop Surg 2019; 11: 176-86. DOI PMID
5.	Li N, Gao J, Mi L, et al. Synovial membrane mesenchymal stem cells: past life, current situation, and application in bone and joint diseases. Stem Cell Res Ther 2020; 11: 381. DOI PMID
6.	Qiong J, Xia Z, Jing L, Haibin W. Synovial mesenchymal stem cells effectively alleviate osteoarthritis through promoting the proliferation and differentiation of meniscus chondrocytes. Eur Rev Med Pharmacol Sci 2020; 24: 1645-55.
7.	Guo S, Tao S, Yin W, Qi X, Sheng J, Zhang C. Exosomes from human synovial-derived mesenchymal stem cells prevent glucocorticoid-induced osteonecrosis of the femoral head in the rat. Int J Biol Sci 2016; 12: 1262-72.
8.	Zhu Y, Wang Y, Zhao B, et al. Comparison of exosomes secreted by induced pluripotent stem cell-derived mesenchymal stem cells and synovial membrane-derived mesenchymal stem cells for the treatment of osteoarthritis. Stem Cell Res Ther 2017; 8: 64. DOI PMID
9.	Tao S, Yuan T, Zhang Y, Yin W, Guo S, Zhang C. Exosomes derived from miR-140-5p-overexpressing human synovial mesenchymal stem cells enhance cartilage tissue regeneration and prevent osteoarthritis of the knee in a rat model. Theranostics 2017; 7: 180-95.
10.	Wang Z, Yan K, Ge G, et al. Exosomes derived from miR-155-5p-overexpressing synovial mesenchymal stem cells prevent osteoarthritis via enhancing proliferation and migration, attenuating apoptosis, and modulating extracellular matrix secretion in chondrocytes. Cell Biol Toxicol 2021; 37: 85-96.
11.	Zeng Z, Dai Y, Deng S, Zou S, Dou T, Wei F. Synovial mesenchymal stem cell-derived extracellular vesicles alleviate chondrocyte damage during osteoarthritis through microRNA-130b-3p-mediated inhibition of the LRP12/AKT/β-catenin axis. Immunopharmacol Immunotoxicol 2022; 44: 247-60.
12.	Zheng T, Li Y, Zhang X, Xu J, Luo M. Exosomes Derived from miR-212-5p overexpressed human synovial mesenchymal stem cells suppress chondrocyte degeneration and inflammation by targeting ELF3. Front Bioeng Biotechnol 2022; 10: 816209.
13.	Qiu M, Liu D, Fu Q. MiR-129-5p shuttled by human synovial mesenchymal stem cell-derived exosomes relieves IL-1β induced osteoarthritis via targeting HMGB1. Life Sci 2021; 269: 118987.
14.	Kong R, Zhang J, Ji L, et al. Synovial mesenchymal stem cell-derived exosomal microRNA-320c facilitates cartilage damage repair by targeting ADAM19-dependent Wnt signalling in osteoarthritis rats. Inflammopharmacology 2023; 31: 915-26. DOI PMID
15.	Wanchen Y, Shijun W, Xuming J, Haijun Z, Xin Z, Yanjiao C. Influence of water decoction of huangqi on immune function in rats with deficiency cold pattern: a study based on transcriptome sequencing technique. Beijing Zhong Yi Yao Da Xue Xue Bao 2018; 41: 759-70.
16.	Zhang X, Song JY, Hu YL, et al. Research progress of the regulation on active compound biosynthesis by the bHLH transcription factors in plants. Yao Xue Xue Bao 2014; 49: 435-42.
17.	Liao Y, Long J, Gallo C, Mirando A, Hilton M. Isolation and culture of murine primary chondrocytes: costal and growth plate cartilage. Methods Mol Biol 2021; 2230: 415-23. DOI PMID
18.	Haseeb A, Lefebvre V. Isolation of mouse growth plate and articular chondrocytes for primary cultures. Methods Mol Biol 2021; 2245: 39-51. DOI PMID
19.	Chen S, Zhou Y, Chen Y, Gu J. Fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics 2018; 34: i884-90.
20.	Langmead B, Salzberg S. Fast gapped-read alignment with Bowtie 2. Nat Methods 2012; 9: 357-9.
21.	Ru Y, Kechris K, Tabakoff B, et al. The multiMiR R package and database: integration of microRNA-target interactions along with their disease and drug associations. Nucleic Acids Res 2014; 42: e133.
22.	Ritchie M, Phipson B, Wu D, et al. Limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res 2015; 43: e47.
23.	Pouran B, Arbabi V, Bleys R, René van Weeren P, Zadpoor A, Weinans H. Solute transport at the interface of cartilage and subchondral bone plate: effect of micro-architecture. J Biomech 2017; 52: 148-54. DOI PMID
24.	Du LL, Yuan PW, Yang W, Li XF, Gao QM. Advantages of micro CT in three-dimensional reconstruction of specimens and its application in animal models of osteoarthritis. Zhong Guo Zu Zhi Gong Cheng Yan Jiu 2022; 26: 1931-6.
25.	Wei Y, Zheng D, Guo X, Zhao M, Gao L, Bai L. Transient receptor potential channel, vanilloid 5, induces chondrocyte apoptosis in a rat osteoarthritis model through the mediation of Ca²⁺ influx. Cell Physiol Biochem 2018; 46: 687-98.
26.	Asghar S, Litherland G, Lockhart J, Goodyear C, Crilly A. Exosomes in intercellular communication and implications for osteoarthritis. Rheumatology (Oxford) 2020; 59: 57-68. DOI PMID
27.	D'Arrigo D, Roffi A, Cucchiarini M, Moretti M, Candrian C, Filardo G. Secretome and extracellular vesicles as new biological therapies for knee osteoarthritis: a systematic review. J Clin Med 2019; 8: 1867.
28.	Ni Z, Kuang L, Chen H, et al. The exosome-like vesicles from osteoarthritic chondrocyte enhanced mature IL-1β production of macrophages and aggravated synovitis in osteoarthritis. Cell Death Dis 2019; 10: 522. DOI PMID
29.	Ding LB, Wang HJ, Li Y, et al. Electroacupuncture stimulating neixiyan (EX-LE5) and dubi (ST35) alleviates osteoarthritis in rats induced by anterior cruciate ligament transaction affecting DNA methylation regulated transcription of miR-146a and miR-140-5p. J Tradit Chin Med 2023; 43: 983-90.
30.	Kopańska M, Szala D, Czech J, et al. MiRNA expression in the cartilage of patients with osteoarthritis. J Orthop Surg Res 2017; 12: 51. DOI PMID
31.	Yang Y, Li P, Zhu S, Bi R. Comparison of early-stage changes of osteoarthritis in cartilage and subchondral bone between two different rat models. PeerJ 2020; 8: e8934.
32.	Wei ZY, Zhang ZL. Interpretation and application of micro-CT to obtain microstructure index in bone metabolism research. Zhong Guo Zu Zhi Gong Cheng Yan Jiu 2018; 11: 200-5.
33.	Snelling S, Bas S, Puskas G, et al. Presence of IL-17 in synovial fluid identifies a potential inflammatory osteoarthritic phenotype. PloS one 2017; 12: e0175109.
34.	Raghu H, Lepus C, Wang Q, et al. CCL2/CCR2, but not CCL5/CCR5, mediates monocyte recruitment, inflammation and cartilage destruction in osteoarthritis. Ann Rheum Dis 2017; 76: 914-22. DOI PMID
35.	Alaaeddine N, Antoniou J, Moussa M, et al. The chemokine CCL20 induces proinflammatory and matrix degradative responses in cartilage. Inflamm Res 2015; 64: 721-31. DOI PMID
36.	Shi X, Ye H, Yao X, Gao Y. The involvement and possible mechanism of NR4A1 in chondrocyte apoptosis during osteoarthritis. Am J Transl Res 2017; 9: 746-54. PMID
37.	Meng J, Ma X, Ma D, Xu C. Microarray analysis of differential gene expression in temporomandibular joint condylar cartilage after experimentally induced osteoarthritis. Osteoarthritis cartilage 2005; 13: 1115-25.
38.	Yang S, Ryu J, Oh H, et al. NAMPT (visfatin), a direct target of hypoxia-inducible factor-2α, is an essential catabolic regulator of osteoarthritis. Ann Rheum Dis 2015; 74: 595-602. DOI PMID
39.	Lai C, Liao B, Peng S, Fang P, Bao N, Zhang L. Synovial fibroblast-miR-214-3p-derived exosomes inhibit inflammation and degeneration of cartilage tissues of osteoarthritis rats. Mol Cell Biochem 2023; 478: 637-49.
40.	Kang L, Yang C, Song Y, et al. MicroRNA-23a-3p promotes the development of osteoarthritis by directly targeting SMAD3 in chondrocytes. Biochem Biophys Res Commun 2016; 478: 467-73.
41.	Dong Y, Li T, Li Y, Ren S, Fan J, Weng X. MicroRNA-23a-3p inhibitor decreases osteonecrosis incidence in a rat model. Mol Med Rep 2017; 16: 9331-6. DOI PMID
42.	Hu H, Dong L, Bu Z, et al. MiR-23a-3p-abundant small extracellular vesicles released from gelma/nanoclay hydrogel for cartilage regeneration. J Extracell Vesicles 2020; 9: 1778883.
43.	Akbaba T, Akkaya-Ulum Y, Tavukcuoglu Z, Bilginer Y, Ozen S, Balci-Peynircioglu B. Inflammation-related differentially expressed common miRNAs in systemic autoinflammatory disorders patients can regulate the clinical course. Clin Exp Rheumatol 2021: 109-17.
44.	Chen Y, Wang Z, Chen X, Peng X, Nie Q. CircNFIC balances inflammation and apoptosis by sponging miR-30e-3p and regulating DENND1B expression. Genes (Basel) 2021; 12: 1829.
45.	Cao Y, Tang S, Nie X, Han W, Zhu Z, Ding C. OP0246 MiR-214-3P protects against osteoarthritis by directly targeting NF-ĸB pathway. Ann Rheum Dis 2020; 79: 155. 2-155.

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