Investigation of gene expression profiles in a rat adjuvant arthritis model suggests an effective role of triptolide via PI3K‑AKT signaling

Zou,Yang; Hu,Weifeng

doi:10.3892/etm.2019.7425

Investigation of gene expression profiles in a rat adjuvant arthritis model suggests an effective role of triptolide via PI3K‑AKT signaling

Authors:
- Yang Zou
- Weifeng Hu
View Affiliations

Affiliations: Department of Orthopedics, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310005, P.R. China
Published online on: March 20, 2019 https://doi.org/10.3892/etm.2019.7425
Pages: 3999-4006
Copyright: © Zou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Rheumatoid arthritis (RA) is a common systemic autoimmune disease mainly involving the formation of a synovial pannus, for which no effective treatment is available. In order to study the molecular biological mechanisms underlying the inhibition of RA synovial pannus by triptolide, differentially expressed genes in synovial tissues from an adjuvant arthritis (AA) rat model with and without triptolide treatment were detected in an mRNA microarray profile produced by Agilent Technologies and verified by reverse transcription‑quantitative polymerase chain reaction analysis (RT‑qPCR). An AA model was established by subcutaneously injecting 0.1 ml Freund's complete adjuvant daily for 18 days and scored by arthritis index assessment. Subsequently, triptolide (0.4 mg/kg) or an equivalent amount of saline was administered daily for 14 days. At the end of the experiment, synovial tissues were obtained from the ankle joints of the rats' hind legs. Total RNA was extracted and purified, and microarray hybridization was used to obtain the gene expression profile for RA with and without triptolide treatment. A total of 48 genes were identified to be differentially expressed between the treatment and model groups, including 32 upregulated and 16 downregulated genes. The possible signaling pathways associated with the effect of triptolide were investigated by Gene Ontology and pathway analysis, revealing that the phosphoinositide‑3 kinase (PI3K)/AKT signaling pathway has a key role in the proliferation and apoptosis of synovial cells in RA joints. Reverse transcription‑quantitative polymerase chain reaction analysis was applied to confirm the aberrant expression of key mRNAs and revealed that vascular endothelial growth factor (VEGF) A and C1q and tumor necrosis factor related protein 3 (C1QTNF3) were downregulated in the treatment group compared with the model group (P<0.05). In conclusion, triptolide may exert its effects against RA via the PI3K/AKT pathway and has an inhibitory effect on the expression of VEGFA and C1QTNF3, thus are potentially associated with the occurrence and development of RA.

View Figures

View References

1	Aletaha D, Neogi T, Silman AJ, Funovits J, Felson DT, Bingham CO III, Birnbaum NS, Burmester GR, Bykerk VP, Cohen MD, et al: 2010 Rheumatoid arthritis classification criteria: An American college of rheumatology/European league against rheumatism collaborative initiative. Arthritis Rheum. 62:2569–2581. 2010. View Article : Google Scholar : PubMed/NCBI
2	Marrelli A, Cipriani P, Liakouli V, Carubbi F, Perricone C, Perricone R and Giacomelli R: Angiogenesis in rheumatoid arthritis: A disease specific process or a common response to chronic inflammation? Autoimmun Rev. 10:595–598. 2011. View Article : Google Scholar : PubMed/NCBI
3	Kong X, Zhang Y, Liu C, Guo W, Li X, Su X, Wan H, Sun Y and Lin N: Anti-angiogenic effect of triptolide in rheumatoid arthritis by targeting angiogenic cascade. PLoS One. 8:e775132013. View Article : Google Scholar : PubMed/NCBI
4	Klareskog L, Catrina AI and Paget S: Rheumatoid arthritis. Lancet. 373:659–672. 2009. View Article : Google Scholar : PubMed/NCBI
5	The Chinese herbal remedy Tripterygium wilfordii Hook F in the treatment of rheumatoid arthritis. Ann Intern Med. 151:I–36. 2009.
6	Goldbach-Mansky R, Wilson M, Fleischmann R, Olsen N, Silverfield J, Kempf P, Kivitz A, Sherrer Y, Pucino F, Csako G, et al: Comparison of Tripterygium wilfordii Hook F versus sulfasalazine in the treatment of rheumatoid arthritis: A randomized trial. Ann Intern Med. 151:229–240, W49-W51. 2009. View Article : Google Scholar : PubMed/NCBI
7	Lu T, Zong M, Fan S, Lu Y, Yu S and Fan L: Thioredoxin 1 is associated with the proliferation and apoptosis of rheumatoid arthritis fibroblast-like synoviocytes. Clin Rheumatol. 37:117–125. 2018. View Article : Google Scholar : PubMed/NCBI
8	Fan D, He X, Bian Y, Guo Q, Zheng K, Zhao Y, Lu C, Liu B, Xu X, Zhang G and Lu A: Triptolide modulates TREM-1 signal pathway to inhibit the inflammatory response in rheumatoid arthritis. Int J Mol Sci. 17:4982016. View Article : Google Scholar : PubMed/NCBI
9	Liu C, Zhang Y, Kong X, Zhu L, Pang J, Xu Y, Chen W, Zhan H, Lu A and Lin N: Triptolide prevents bone destruction in the collagen-induced arthritis model of rheumatoid arthritis by targeting RANKL/RANK/OPG signal pathway. Evid Based Complement Alternat Med. 2013:6260382013.PubMed/NCBI
10	Lin N, Liu C, Xiao C, Jia H, Imada K, Wu H and Ito A: Triptolide, a diterpenoid triepoxide, suppresses inflammation and cartilage destruction in collagen-induced arthritis mice. Biochem Pharmacol. 73:136–146. 2007. View Article : Google Scholar : PubMed/NCBI
11	Chen BJ: Triptolide, a novel immunosuppressive and anti-inflammatory agent purified from a Chinese herb Tripterygium wilfordii Hook F. Leuk Lymphoma. 42:253–265. 2001. View Article : Google Scholar : PubMed/NCBI
12	Glehr M, Fritsch-Breisach M, Lohberger B, Walzer SM, Moazedi-Fuerst F, Rinner B, Gruber G, Graninger W, Leithner A and Windhager R: Influence of resveratrol on rheumatoid fibroblast-like synoviocytes analysed with gene chip transcription. Phytomedicine. 20:310–318. 2013. View Article : Google Scholar : PubMed/NCBI
13	Li ZJ, Wang DQ, Hu SP, Wang XF, He Y and Zhang ZJ: A gene chip study of ‘Jun Du Yan Bingzhi’ on liver in a sepsis model of rat. Zhongguo Wei Zhong Bing Ji Jiu Yi Xue. 21:44–47. 2009.(In Chinese). PubMed/NCBI
14	Collins JF: Gene chip analyses reveal differential genetic responses to iron deficiency in rat duodenum and jejunum. Biol Res. 39:25–37. 2006.PubMed/NCBI
15	Saeki Y, Matsui T, Saisho K and Tohma S: Current treatments of rheumatoid arthritis: From the ‘NinJa’ registry. Expert Rev Clin Immunol. 8:455–465. 2012. View Article : Google Scholar : PubMed/NCBI
16	Wang Y, Wei D, Lai Z and Le Y: Triptolide inhibits CC chemokines expressed in rat adjuvant-induced arthritis. Int Immunopharmacol. 6:1825–1832. 2006. View Article : Google Scholar : PubMed/NCBI
17	Yifan W, Dengming W, Zheng L, Yanping L and Junkan S: Triptolide inhibits CCR5 expressed in synovial tissue of rat adjuvant-induced arthritis. Pharmacol Rep. 59:795–799. 2007.PubMed/NCBI
18	Li S, Chen JW, Xie X, Tian J, Deng C, Wang J, Gan HN and Li F: Autophagy inhibitor regulates apoptosis and proliferation of synovial fibroblasts through the inhibition of PI3K/AKT pathway in collagen-induced arthritis rat model. Am J Transl Res. 9:2065–2076. 2017.PubMed/NCBI
19	Xiao C, Zhou J, He Y, Jia H, Zhao L, Zhao N and Lu A: Effects of triptolide from Radix Tripterygium wilfordii (Leigongteng) on cartilage cytokines and transcription factor NF-kappaB: A study on induced arthritis in rats. Chin Med. 4:132009. View Article : Google Scholar : PubMed/NCBI
20	Jiao Y, Ding H, Huang S, Liu Y, Sun X, Wei W, Ma J and Zheng F: Bcl-XL and Mcl-1 upregulation by calreticulin promotes apoptosis resistance of fibroblast-like synoviocytes via activation of PI3K/Akt and STAT3 pathways in rheumatoid arthritis. Clin Exp Rheumatol. 36:841–849. 2018.PubMed/NCBI
21	Zhou J, Xiao C, Zhao L, Jia H, Zhao N, Lu C, Yang D, Tang JC, Chan AS and Lu AP: The effect of triptolide on CD4+ and CD8+ cells in Peyer's patch of SD rats with collagen induced arthritis. Int Immunopharmacol. 6:198–203. 2006. View Article : Google Scholar : PubMed/NCBI
22	Evans SJ, Datson NA, Kabbaj M, Thompson RC, Vreugdenhil E, De Kloet ER, Watson SJ and Akil H: Evaluation of affymetrix gene chip sensitivity in rat hippocampal tissue using SAGE analysis. Serial analysis of gene expression. Eur J Neurosci. 16:409–413. 2002. View Article : Google Scholar : PubMed/NCBI
23	Wang J, Jiang Z, Ji J, Wang X, Wang T, Zhang Y, Tai T, Chen M, Sun L, Li X and Zhang L: Gene expression profiling and pathway analysis of hepatotoxicity induced by triptolide in Wistar rats. Food Chem Toxicol. 58:495–505. 2013. View Article : Google Scholar : PubMed/NCBI
24	Wang J, Wang A, Zeng H, Liu L, Jiang W, Zhu Y and Xu Y: Effect of triptolide on T-cell receptor beta variable gene mRNA expression in rats with collagen-induced arthritis. Anat Rec (Hoboken). 295:922–927. 2012. View Article : Google Scholar : PubMed/NCBI
25	Qu Y, Wu J, Deng JX, Zhang YP, Liang WY, Jiang ZL, Yu QH and Li J: MicroRNA-126 affects rheumatoid arthritis synovial fibroblast proliferation and apoptosis by targeting PIK3R2 and regulating PI3K-AKT signal pathway. Oncotarget. 7:74217–74226. 2016. View Article : Google Scholar : PubMed/NCBI
26	Wang M, Chen DQ, Chen L, Liu D, Zhao H, Zhang ZH, Vaziri ND, Guo Y, Zhao YY and Cao G: Novel RAS inhibitors poricoic acid ZG and poricoic acid ZH attenuate renal fibrosis via a Wnt/β-catenin pathway and targeted phosphorylation of smad3 signaling. J Agric Food Chem. 66:1828–1842. 2018. View Article : Google Scholar : PubMed/NCBI
27	Ramirez-Bello J, Cadena-Sandoval D, Fragoso JM, Barbosa-Cobos RE, Moreno-Eutímio MA, Saavedra-Salinas MÁ, Valencia-Pacheco G, López-Villanueva RF and Jiménez-Morales S: The VEGFA-1154G/A polymorphism is associated with reduced risk of rheumatoid arthritis but not with systemic lupus erythematosus in Mexican women. J Gene Med. 20:e30242018. View Article : Google Scholar : PubMed/NCBI
28	Murayama MA, Kakuta S, Maruhashi T, Shimizu K, Seno A, Kubo S, Sato N, Saijo S, Hattori M and Iwakura Y: CTRP3 plays an important role in the development of collagen-induced arthritis in mice. Biochem Biophys Res Commun. 443:42–48. 2014. View Article : Google Scholar : PubMed/NCBI
29	Salliot C and van der Heijde D: Long-term safety of methotrexate monotherapy in patients with rheumatoid arthritis: A systematic literature research. Ann Rheum Dis. 68:1100–1104. 2009. View Article : Google Scholar : PubMed/NCBI
30	Malemud CJ: Negative regulators of JAK/STAT signaling in rheumatoid arthritis and osteoarthritis. Int J Mol Sci. 18:4842017. View Article : Google Scholar
31	Wang J, Li Y, Yang Y, Du J, Zhao M, Lin F, Zhang S and Wang B: Systems pharmacology dissection of multiscale mechanisms of action for herbal medicines in treating rheumatoid arthritis. Mol Pharm. 14:3201–3217. 2017. View Article : Google Scholar : PubMed/NCBI
32	Tu S, Hu Y, Zeng K, Zhang M, Lai X and Weichen Z: Effects of triptolide on the expression and activity of NF-kappaB in synovium of collagen-induced arthritis rats. J Huazhong Univ Sci Technolog Med Sci. 25:543–545. 2005. View Article : Google Scholar : PubMed/NCBI
33	Matta R, Wang X, Ge H, Ray W, Nelin LD and Liu Y: Triptolide induces anti-inflammatory cellular responses. Am J Transl Res. 1:267–282. 2009.PubMed/NCBI
34	Yang M, Huang J, Pan HZ and Jin J: Triptolide overcomes dexamethasone resistance and enhanced PS-341-induced apoptosis via PI3k/Akt/NF-kappaB pathways in human multiple myeloma cells. Int J Mol Med. 22:489–496. 2008.PubMed/NCBI
35	Liu YY, Jiao ZY, Li W and Tian Q: PI3K/AKT signaling pathway activation in a rat model of migraine. Mol Med Rep. 16:4849–4854. 2017. View Article : Google Scholar : PubMed/NCBI
36	Clarimundo VS, Farinon M, Pedó RT, Teixeira VON, Nör C, Gulko PS, Xavier RM and de Oliveira PG: Gastrin-releasing peptide and its receptor increase arthritis fibroblast-like synoviocytes invasiveness through activating the PI3K/AKT pathway. Peptides. 95:57–61. 2017. View Article : Google Scholar : PubMed/NCBI