PPAR Research

PPAR Research / 2019 / Article

Research Article | Open Access

Volume 2019 |Article ID 6403504 | https://doi.org/10.1155/2019/6403504

Weiyan Wang, Chunling Li, Zhiyi Zhang, Yue Zhang, "Arsenic Trioxide in Synergy with Vitamin D Rescues the Defective VDR-PPAR- Functional Module of Autophagy in Rheumatoid Arthritis", PPAR Research, vol. 2019, Article ID 6403504, 11 pages, 2019. https://doi.org/10.1155/2019/6403504

Arsenic Trioxide in Synergy with Vitamin D Rescues the Defective VDR-PPAR- Functional Module of Autophagy in Rheumatoid Arthritis

Academic Editor: Antonio Brunetti
Received20 Nov 2018
Revised17 Mar 2019
Accepted01 Apr 2019
Published07 May 2019


Dysregulated autophagy leads to autoimmune diseases including rheumatoid arthritis (RA). Arsenic trioxide (ATO) is a single agent used for the treatment of acute promyelocytic leukemia and is highly promising for other malignancies but is also attractive for RA, although its relationship with autophagy remains to be further clarified and its application optimized. For the first time, we report a defective functional module of autophagy comprising the Vitamin D receptor (VDR), PPAR-γ, microtubule-associated protein 1 light-chain 3 (LC3), and p62 which appears in RA synovial fibroblasts. ATO alleviated RA symptoms by boosting effective autophagic flux through significantly downregulating p62, the inflammation and catabolism protein. Importantly, low-dose ATO synergizes with Vitamin D in RA treatment.

1. Introduction

Rheumatoid arthritis (RA) is a chronic inflammatory joint disease that leads to cartilage and bone damage as well as disability [1].

Autophagy is defined as a degradation mechanism by which cells recycle cytoplasmic components. Several autophagy-related genes are involved including Beclin1, microtubule-associated protein 1 light-chain 3 (LC-3), p62, and mammalian target of rapamycin (mTOR) [24]. The assessment of autophagic activity can be achieved by detection of the LC-3 protein. Autophagic flux is a complex process that involves transporting, binding, degrading, and recycling the cytoplasmic components. Nevertheless, increases in the levels of LC-3 can be caused by either the induction of autophagy or inhibited fusion of the autophagosomes with lysosomes; it cannot be used to monitor autophagic flux per se. The p62 protein serves as a link between LC-3 and ubiquitinated substrates and is degraded in autolysosomes. Thus, inhibition of the final step of autophagy correlates with increased levels of p62 and indicates impaired flux [5]. Dysregulated autophagic flux is involved in the pathogenesis of many autoimmune diseases, including RA, by regulating the organism’s lifespan and cartilage homeostasis [69]. In addition, the activation of autophagy in immune cells is significantly associated with inflammatory parameters such as interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), which are proinflammatory cytokines involved in the pathogenesis and progression of RA [10].

The Vitamin D receptor (VDR) is expressed in multiple cells and numerous RA and autophagy-related genes, including mTOR, are potential candidate targets of Vitamin D (Vit D) and VDR [11, 12]. It maintains “robustness” in unfavorable conditions and serves a “capacitor” in homeostasis in RA biology [13, 14]. Peroxisome proliferator-activated receptor-γ (PPAR-γ) is a primary target of Vit D [15]. In addition, both VDR and PPAR-γ are key autophagy regulators [16, 17] and play central roles in the development and progression of RA, supporting the possible involvement of autophagy in RA. In vivo and in vitro experiments confirmed that the PPAR-γ agonist could downregulate inflammatory cytokines [18, 19]. Bone erosion can be alleviated in RA patients by reducing catabolism through PPAR-γ pathway activation [20]. The regulatory effects of VDR and PPAR-γ through autophagy have been proven in tumors and other diseases [2123]. Nevertheless, the exact role and involvement of VDR and PPAR-γ in RA-related autophagy remain to be defined.

The search for novel drugs that can manage more than one single age-related disease is encouraged [24]. Among these drugs, arsenic trioxide (As2O3, ATO) is recognized for treating tumors [25, 26] and autoimmune rheumatic diseases by enhancing apoptosis and inhibiting angiogenesis [2729]. ATO has been shown to induce antitumor effects through autophagy [30]. However, the effect of ATO on autophagy in RA is unknown.

In the present study, we demonstrated for the first time that VDR, PPAR-γ, and LC-3 participate in a functional module of autophagy (a functional module is a group of genes that are tightly associated through multiple feedback loops) and are significantly upregulated in RA synovial tissues, although autophagic flux was unexpectedly severely impaired. Surprisingly, ATO rescued this defective functional module of autophagy in RA fibroblast-like synoviocytes (FLS) and mice with RA, and the effect was even better when ATO was used with Vit D. Hence, ATO combined with Vit D could be a potential therapeutic strategy against RA.

2. Materials and Methods

2.1. Synovial Fibroblast Culture and TNF-α Stimulation

RA FLS and normal human (NH) FLS were purchased from Cell Applications (San Diego, CA, USA) and maintained in a synoviocyte growth medium (Cell Applications). Cells were used for the experiments in stages 4–6. RA FLS were pretreated with 50 ng/mL of TNF-α for 4 h before the application of ATO and/or Vit D. Cell culture supernatants were used for enzyme-linked immunosorbent assays (ELISAs) 48 h after the addition of the treatments.

2.2. In Vitro Proliferation Assay

Cell proliferation was evaluated with a Cell Counting Kit-8 (Sigma, St Louis, MO, USA) following procedures described earlier with minor modifications [31]. Briefly, 104 cells were seeded in a 96-well plate. After 24 h, different concentrations of drugs or vehicles were added with fresh medium. Cells were incubated at 37°C for 48 h. The plates were read at 450 nm. The experiments were repeated three times.

2.3. RNA Preparation and Real-Time Quantitative Polymerase Chain Reaction Analysis

Total RNA was extracted from FLS with TRIzol reagent (Invitrogen, Carlsbad, CA, USA) and converted to cDNA. Real-time quantitative polymerase chain reaction (PCR) amplification was performed as described previously [28]. The sequences of the primers are shown in Table 1. GAPDH was used as an internal control. All samples were measured in triplicate and the results were evaluated via the method [9].

Target geneForward primer (5′–3′)Reverse primer (5′–3′)


2.4. Western Blotting

Equal amounts of proteins were obtained and separated by SDS-PAGE and transferred to a polyvinylidene fluoride membrane as described previously [28]. Specific primary antibodies were added, including anti-PPAR-γ (catalog no. ab41928, Abcam, Cambridge, MA, USA), anti-VDR (catalog no. 12550, Cell Signaling, Cambridge, MA, USA), anti-LC-3-I/II (catalog no. 12741, Cell Signaling), anti-p62 (catalog no. ab56416, Abcam), anti-mTOR (catalog no. ab32028, Abcam), and anti-p-mTOR (catalog no. ab109268, Abcam). The reactive bands were visualized with a chemiluminescence detection system and analyzed with ImageJ software (National Institutes of Health, USA).

2.5. RNA Interference

VDR knockdown was achieved by transfecting specific VDR small interfering RNA (siRNA) into RA FLS. The siRNA sequence was as follows: forward, 5′-GCUGAAGUCAAGUGCCAUUTT-3′; reverse, 5′-AAUGGCACUUGACUUCAGCTT-3′. RA FLS were plated in 12-well plates at 105 cells per well with serum-free DMEM and transfected with siRNA via Lipofectamine 2000 (Invitrogen) as previously described [28]. Six hours after transfection, the culture medium was replaced by DMEM with 10% FBS. Knockdown efficiency was determined by real-time PCR and western blot analysis. The experiments were repeated at least three times.

2.6. Establishment of Collagen-Induced Arthritis

Specific pathogen-free 6-week-old male DBA/1J mice weighing 18±2 g were purchased from SLAC (Shanghai, China). Animal welfare and experimental procedures were carried out in accordance with the Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee of Harbin Medical University. The collagen-induced arthritis (CIA) mouse model was established as described previously [28]. Mice were randomly assigned to different groups (n=6 per group): normal control group (mice without immunization and injected with saline); the CIA control group (CIA mice treated with saline, CIA-saline); the ATO-treated group (CIA mice treated with ATO at a dose of 1.0 or 2.0 mg/kg/day), Vit D (400 ng/kg/d) or Vit D (400 ng/kg/d) + ATO (2.0 mg/kg/day); and the methotrexate (MTX) group (CIA mice treated with MTX at 2 mg/kg/week as a positive control). Mice were given intraperitoneal injections from day 28 to day 41. The body weight of each mouse was recorded every other day from day 21.

2.7. Assessment of Arthritis Severity

To evaluate the severity of the arthritis quantitatively, the arthritis score was assessed every 2 days starting from day 21, according to a scoring system used previously [32]. In addition, body weight and the thickness of the two hind paws were measured every other day.

2.8. Microcomputed Tomography Imaging

To investigate the effects of ATO on the three-dimensional (3D) bone structure, we conducted an assay using microcomputed tomography (micro-CT) imaging (Quantum GX, Perkin Elmer, Waltham, USA). Mice were scanned and reconstructed into a 3D structure via micro-CT imaging 39 days after the initial collagen injection. The voxel size was 72 μm, the X-ray tube voltage was 90 KV, the current was 88 μA, and the exposure time was 4 min. Mean CT values of the hind paws were calculated with Caliper Analyze software (Analyze Direct, Kansas, USA) to assess bone loss.

2.9. Histological Analysis

Whole knee joints of the mice were collected and fixed in 10% buffered neutral formalin and decalcified in 10% EDTA for 4 weeks. The paraffin-embedded specimens were stained with hematoxylin and eosin (H&E). Histological changes were scored in a blinded manner by two independent observers based on synovial hyperplasia, joint inflammation, and bone erosion following a scoring system described previously [33, 34].

2.10. Immunohistochemistry Analysis

Immunohistochemistry was performed with specific antibodies for target proteins following a protocol described previously with some modifications [6]. Knee joint sections on slides were incubated with anti-VDR, anti-PPAR-γ, anti-LC-3, or anti-P62 (Boster, Wuhan, China) antibodies. Subsequently, the sections were stained with a polymer horseradish peroxidase (HRP) detection system (PV9001, ZSGB-BIO, Beijing, China) and visualized with a diaminobenzidine (DAB) peroxidase substrate kit (ZLI-9017, ZSGB-BIO, Beijing, China). Each section was evaluated under a microscope (DMi8, LEICA, Wetzlar, Germany) in three randomly selected areas at a magnification of 20×. Image-Pro Plus 6 (Media Cybernetics, Inc.) was used to analyze the average integrated optical density (IOD) according to a previously described protocol [35].

2.11. Enzyme-Linked Immunosorbent Assay

Serum samples from the mice and the cell culture supernatant were collected and stored at −80°C. The concentrations of IL-6, IL-1β, matrix metalloproteinase-3 (MMP-3) and MMP-13 were detected with commercial kits (Elabscience Biotechnology Co., Ltd., Wuhan, China), according to the manufacturer’s protocols. All assays were conducted in triplicate.

2.12. Statistical Analysis

Data are represented as means ± standard error of the mean and were analyzed via Student’s t-test or analysis of variance (ANOVA), as appropriate. All analyses were carried out in SPSS 17.0 software. Values of p < 0.05 were considered statistically significant.

3. Results

3.1. ATO Rescues the Defective VDR-PPAR-γ Functional Module of Autophagy Both In Vivo and In Vitro

Regarding potential side effects and safety, we examined the influence of ATO and Vit D on the proliferation of RA FLS. Although the optical density value decreased, different concentrations of ATO and Vit D showed no significant influence on cell proliferation (Figure 1(a)). RA FLS were detected after treatment with different doses of ATO (0.1, 0.5, 1.0, 2.0, and 4.0 μM). VDR, PPAR-γ, and LC-3 were significantly upregulated after ATO treatment, with the peak effect seen at 2 μM; importantly, p62 was surprisingly significantly downregulated (Figure 2(a), p<0.05). Whether or not RA FLS exhibited defective autophagic flux was investigated by using VDR, PPAR-γ, LC-3, and p62 mRNA and protein in normal human (NH) FLS and RA FLS. The expression levels of VDR, PPAR-γ, and LC-3 were significantly upregulated in RA FLS compared with normal human FLS. Meanwhile, p62 mRNA and protein were also higher in RA FLS, which may indicate blockage of autophagy (Figure 1(b), p <0.05).

Afterwards, RA FLS were treated with ATO (2 μM) after VDR knockdown. PPAR-γ and LC-3 were significantly downregulated but p62 was enhanced after silencing VDR, and ATO could reverse these effects (Figure 1(c), p<0.05). Immunohistochemistry was performed to validate the effect of ATO on the VDR-PPAR-γ autophagy functional module in CIA mice. Strikingly, treatment with ATO (2 mg/kg/d) or MTX (2 mg/kg/week) led to significant increases in the expression of VDR, PPAR-γ, and LC-3 and a decrease in p62 compared with CIA-saline mice (Figure 2(b), p<0.05). Thus, ATO alone may activate the VDR-PPAR-γ autophagy functional module both in vitro and in vivo.

3.2. Combined Effect of ATO and Vit D on the VDR-PPAR-γ Functional Module of Autophagy

Both ATO and Vit D upregulated the expression of VDR, PPAR-γ, and LC-3 in vitro (Figure 3(a), p<0.05), but ATO had a synergistic effect with Vit D on this upregulation in RA FLS (Figure 3(a), p<0.05); low doses of ATO (1 mg/kg/d) plus Vit D (400 ng/kg/d) achieved similar effects to ATO (2 mg/kg/d) (Figures 3(a) and 4(b)). Furthermore, ATO synergized with Vit D in vivo to upregulate the expression of VDR, PPAR-γ, and LC-3 significantly and downregulate p62 in the synovium of mice (Figure 2(b), p<0.05). Therefore, we consider that ATO may have a synergistic effect with Vit D on the VDR-PPAR-γ autophagy functional module in vivo and in vitro.

3.3. ATO Alleviates Symptoms and Joint Destruction in CIA Mice

We assessed the body weight, degree of paw swelling, and arthritis scores of the CIA and control mice. Both ATO (1 or 2 mg/kg/d) and Vit D (400 ng/kg/d) alone alleviated the symptoms of CIA mice (Figure 5(c), p<0.01). However, combining ATO and Vit D further reduced the arthritis score but this was not significantly different from ATO alone. The severity of arthritis in CIA mice was assessed by H&E staining of knee joint sections. ATO treatment at 2 mg/kg/d and Vit D (400 ng/kg/d) significantly reduced the histological scores including synovial hyperplasia, cartilage, and bone erosion and joint inflammatory (Figure 5(b), n=6, p<0.01). Additionally, ATO (2 mg/kg/d) significantly enhanced the mean CT values of hind paws compared with CIA-saline mice, as did MTX (2 mg/kg/w) (Figure 5(a); n=6, p<0.01).

3.4. ATO Inhibits the TNF-α-Induced Inflammation Module Representative and Catabolism Module Representative Release by Regulating the VDR-PPAR-γ Autophagy Functional Module

Interestingly, ATO (1 mg/kg/d) decreased MMP-13 expression, but key inflammation biomarkers such as IL-6, IL-1β, and IL-8 and catabolism factor module representatives including MMP-3 and MMP-13 were significantly suppressed by ATO (2 mg/kg/d) (Figure 4(a), p<0.01) but were significantly upregulated in CIA-saline mice compared with the normal control mice, as shown by ELISA (Figure 4(a), p<0.01). Vit D (400 ng/kg/d) alone significantly decreased these cytokines and may significantly enhance the effect of ATO when used in combination (Figure 4(a), p<0.05). Therefore, ATO and Vit D downregulated inflammation factors and catabolism factors in CIA mouse serum in vivo.

Furthermore, TNF-α treatment (50 ng/mL) significantly induced the inflammation factors and catabolism factors in RA FLS. Stimulation of the VDR-PPAR-γ functional module by its agonist Vit D and rosiglitazone (50 μmol/L) decreased cytokine release, as did ATO with or without Vit D. In contrast, silencing VDR by siRNA and inhibiting PPAR-γ with GW9662 (10 μmol/L) increased the secretion of inflammatory and catabolic factors (Figure 4(b), p<0.05); this was reversed by ATO. Furthermore, inflammation and catabolism factors were downregulated by stimulating autophagy with rapamycin (100 nmol/L). Consistently, the autophagy blocker BafA1 induced cytokine release but ATO reversed this effect (Figure 4(b), p<0.05). Therefore, ATO inhibited the release of TNF-α-induced inflammation factors and catabolism factors in RA FLS by regulating the VDR-PPAR-γ autophagy module.

4. Discussion

Autophagy is a highly conserved biological process in eukaryotic cells. Some autophagy-associated genes are closely related to RA [36].

Firstly, we explored the roles of VDR and PPAR-γ during autophagy in RA FLS. To the best of our knowledge, this is the first study reporting that the expressions of VDR, PPAR-γ, and LC-3 are significantly upregulated, and mTOR is significantly downregulated in RA FLS. Our results showed that p62 in RA FLS was significantly upregulated, indicating autophagic flux obstruction. Since impaired autophagic flux may be involved in a variety of diseases [37], we concluded that defective autophagic flux, rather than other factors causing functional loss of autophagy, leads to RA.

Indeed, PPAR-γ and VDR are interconnected and modulate the expression of genes in similar ways as a functional module [38, 39]. In order to further clarify the relationship between PPAR-γ and VDR in RA FLS, we applied rosiglitazone to excite PPAR-γ plaques in RA FLS and found that VDR expression was also elevated, suggesting that PPAR-γ may have a positive feedback effect on VDR.

Anti-TNF biological agents have been proven to be effective in the treatment of RA; however, around 30% of the patients respond poorly or not at all to the biologics. In addition, the treatment is limited by preexisting malignancy and inflammation and further impose an economic burden on patients because of the high costs. ATO has attracted global attention as one of several highly promising effective anticancer drugs. To our knowledge, this is the first report to demonstrate that ATO may rescue impaired autophagic flux beyond its role in activating the VDR-PPAR-γ autophagy functional module, consequently inhibiting the inflammatory response and joint destruction.

Previously, we and others reported that ATO may relieve the clinical symptoms of patients with leukemia and other rheumatic diseases [27, 40, 41]. Although we observed no significant effect of ATO on cell proliferation and no toxic effect on CIA mice, several adverse effects have been reported in patients such as liver injury and carcinogenesis [41], which limit the application of ATO in clinical settings. Solution could be sought regarding local drug delivery, drug combinations [42], or chemical modifications [43]. For the first time, we provided evidence that ATO showed a synergistic effect in combination with Vit D both in vivo and in vitro. Since Vit D is safe and easily available, the combination of the two drugs may reduce the dosage of ATO and mitigate its side effects.

5. Conclusion

In closing, VDR, PPAR-γ, LC-3, and p62 form a functional module, which is obviously upregulated in RA FLS and CIA mice synovial fibroblasts, but with impaired autophagic flux, as demonstrated by the upregulation of p62.

Although the exact mechanisms are still poorly understood, ATO may enhance the VDR-PPAR-γ autophagy functional module and rescue impaired autophagic flux both in vivo and in vitro in RA and consequently inhibit the expression of inflammatory and catabolic factors that participate in joint destruction (Figure 6). Furthermore, we provided evidence that low-dose ATO showed a better effect in combination with Vit D in ameliorating RA symptoms, suggesting novel promising protocols for RA and cancer therapy.

Data Availability

The data used to support the findings of this study are available from the corresponding author upon request.

Conflicts of Interest

The authors declare that they have no conflicts of interest.

Authors’ Contributions

Weiyan Wang, Chun Ling Li, and Yue Zhang contributed equally to this work.


This work is supported by grants from the National Natural Science Foundation of China to Zhiyi Zhang (NSFC 81771749) and Yue Zhang (NSFC 81771748).


  1. J. S. Smolen, D. Aletaha, and I. B. McInnes, “Rheumatoid arthritis,” The Lancet, vol. 388, no. 10055, pp. 2023–2038, 2016. View at: Publisher Site | Google Scholar
  2. J. S. Rockel and M. Kapoor, “Autophagy: controlling cell fate in rheumatic diseases,” Nature Reviews Rheumatology, vol. 12, no. 9, pp. 517–531, 2016. View at: Publisher Site | Google Scholar
  3. A. M. Choi, S. W. Ryter, and B. Levine, “Autophagy in human health and disease,” The New England Journal of Medicine, vol. 368, no. 7, pp. 651–662, 2013. View at: Publisher Site | Google Scholar
  4. A. Doria, M. Gatto, and L. Punzi, “Autophagy in human health and disease,” The New England Journal of Medicine, vol. 368, no. 19, pp. 1845-1846, 2013. View at: Publisher Site | Google Scholar
  5. B. Levine and G. Kroemer, “Biological functions of autophagy genes: a disease perspective,” Cell, vol. 176, pp. 11–42, 2019. View at: Google Scholar
  6. F. Vasheghani, Y. Zhang, Y.-H. Li et al., “PPARgamma deficiency results in severe, accelerated osteoarthritis associated with aberrant mTOR signalling in the articular cartilage,” Annals of the Rheumatic Diseases, vol. 74, no. 3, pp. 569–578, 2015. View at: Publisher Site | Google Scholar
  7. D. E. Harrison, R. Strong, Z. D. Sharp et al., “Rapamycin fed late in life extends lifespan in genetically heterogeneous mice,” Nature, vol. 460, no. 7253, pp. 392–395, 2009. View at: Publisher Site | Google Scholar
  8. T. Vellai, K. Takacs-Vellai, Y. Zhang, A. L. Kovacs, L. Orosz, and F. Müller, “Genetics: influence of TOR kinase on lifespan in C. elegans,” Nature, vol. 426, no. 6967, pp. 620-621, 2003. View at: Google Scholar
  9. Y. Zhang, F. Vasheghani, Y.-H. Li et al., “Cartilage-specific deletion of mTOR upregulates autophagy and protects mice from osteoarthritis,” Annals of the Rheumatic Diseases, vol. 74, no. 7, pp. 1432–1440, 2015. View at: Publisher Site | Google Scholar
  10. Y. Matsuzawa-Ishimoto, S. Hwang, and K. Cadwell, “Autophagy and Inflammation,” Annual Review of Immunology, vol. 36, pp. 73–101, 2018. View at: Publisher Site | Google Scholar
  11. Y. Okada, D. Wu, G. Trynka, T. Raj et al., “Genetics of rheumatoid arthritis contributes to biology and drug discovery,” Nature, vol. 506, pp. 376–381, 2014. View at: Google Scholar
  12. S. V. Ramagopalan, A. Heger, A. J. Berlanga et al., “A ChIP-seq defined genome-wide map of vitamin D receptor binding: associations with disease and evolution,” Genome Research, vol. 20, no. 10, pp. 1352–1360, 2010. View at: Publisher Site | Google Scholar
  13. Y. Zhang, “Emerging vitamin D receptor-centered patterns of genetic overlap across some autoimmune diseases and associated cancers,” Journal of Genetic Syndromes & Gene Therapy, vol. 04, no. 11, Article ID 1000e1123, 2013. View at: Publisher Site | Google Scholar
  14. D. Hochbaum, Y. Zhang, C. Stuckenholz et al., “DAF-12 regulates a connected network of genes to ensure robust developmental decisions,” PLoS Genetics, vol. 7, no. 7, Article ID e1002179, 2011. View at: Google Scholar
  15. F. Alimirah, X. Peng, L. Yuan et al., “Crosstalk between the peroxisome proliferator-activated receptor gamma (PPARγ) and the vitamin D receptor (VDR) in human breast cancer cells: PPARγ binds to VDR and inhibits 1α,25-dihydroxyvitamin D3 mediated transactivation,” Experimental Cell Research, vol. 318, no. 19, pp. 2490–2497, 2012. View at: Publisher Site | Google Scholar
  16. T. Yao, X. Ying, Y. Zhao et al., “Vitamin D receptor activation protects against myocardial reperfusion injury through inhibition of apoptosis and modulation of autophagy,” Antioxidants & Redox Signaling, vol. 22, no. 8, pp. 633–650, 2015. View at: Publisher Site | Google Scholar
  17. L. Wang, Y. Yin, G. Hou, J. Kang, and Q. Wang, “Peroxisome proliferator-activated receptor (PPARγ) plays a protective role in cigarette smoking-induced inflammation via AMP-activated protein kinase (AMPK) signaling,” Medical Science Monitor, vol. 24, pp. 5168–5177, 2018. View at: Publisher Site | Google Scholar
  18. L. F. da Rocha Junior, M. J. Rêgo, and M. B. Cavalcanti, “Synthesis of a novel thiazolidinedione and evaluation of its modulatory effect on IFN-γ, IL-6, IL-17A, and IL-22 production in PBMCs from rheumatoid arthritis Patients,” BioMed Research International, vol. 2013, Article ID 926060, 8 pages, 2013. View at: Publisher Site | Google Scholar
  19. V. Costa, D. Foti, F. Paonessa et al., “The insulin receptor: a new anticancer target for peroxisome proliferator-activated receptor-gamma (PPARgamma) and thiazolidinedione-PPARgamma agonists,” Endocrine-Related Cancer, vol. 15, no. 1, pp. 325–335, 2008. View at: Publisher Site | Google Scholar
  20. M. Koufany, D. Chappard, P. Netter et al., “The peroxisome proliferator-activated receptor gamma agonist pioglitazone preserves bone microarchitecture in experimental arthritis by reducing the interleukin-17-dependent osteoclastogenic pathway,” Arthritis & Rheumatism, vol. 65, no. 12, pp. 3084–3095, 2013. View at: Publisher Site | Google Scholar
  21. L. E. Tavera-Mendoza, T. Westerling, E. Libby et al., “Vitamin D receptor regulates autophagy in the normal mammary gland and in luminal breast cancer cells,” Proceedings of the National Acadamy of Sciences of the United States of America, vol. 114, no. 11, pp. E2186–E2194, 2017. View at: Publisher Site | Google Scholar
  22. L. M. Das, A. M. Binko, Z. P. Traylor, H. Peng, and K. Q. Lu, “Vitamin D improves sunburns by increasing autophagy in M2 macrophages,” Autophagy, vol. 15, no. 5, pp. 813–826, 2019. View at: Publisher Site | Google Scholar
  23. J. Zhong, W. Gong, J. Chen et al., “Micheliolide alleviates hepatic steatosis in db/db mice by inhibiting inflammation and promoting autophagy via PPAR-γ-mediated NF-кB and AMPK/mTOR signaling,” International Immunopharmacology, vol. 59, pp. 197–208, 2018. View at: Publisher Site | Google Scholar
  24. I. Bellantuono, “Find drugs that delay many diseases of old age,” Nature, vol. 554, no. 7692, pp. 293–295, 2018. View at: Publisher Site | Google Scholar
  25. Z. Chen and S.-J. Chen, “Poisoning the Devil,” Cell, vol. 168, no. 4, pp. 556–560, 2017. View at: Publisher Site | Google Scholar
  26. H. Zhang, L. Yang, J. Ling et al., “Systematic identification of arsenic-binding proteins reveals that hexokinase-2 is inhibited by arsenic,” Proceedings of the National Acadamy of Sciences of the United States of America, vol. 112, no. 49, pp. 15084–15089, 2015. View at: Publisher Site | Google Scholar
  27. Y. Mei, Y. Zheng, H. Wang et al., “Arsenic trioxide induces apoptosis of fibroblast-like synoviocytes and represents antiarthritis effect in experimental model of rheumatoid arthritis,” The Journal of Rheumatology, vol. 38, no. 1, pp. 36–43, 2011. View at: Publisher Site | Google Scholar
  28. J. Zhang, C. Li, Y. Zheng, Z. Lin, Y. Zhang, and Z. Zhang, “Inhibition of angiogenesis by arsenic trioxide via TSP-1–TGF-β1-CTGF–VEGF functional module in rheumatoid arthritis,” Oncotarget, vol. 8, no. 43, pp. 73529–73546, 2017. View at: Google Scholar
  29. Y. Zhao, G. Wen, Z. Qiao et al., “Effects of tetra-arsenic tetra-sulfide on BXSB lupus-prone mice: A pilot study,” Lupus, vol. 22, no. 5, pp. 469–476, 2013. View at: Publisher Site | Google Scholar
  30. T. Li, R. Ma, Y. Zhang et al., “Arsenic trioxide promoting ETosis in acute promyelocytic leukemia through mTOR-regulated autophagy,” Cell Death & Disease, vol. 9, no. 2, pp. 1–14, 2018. View at: Publisher Site | Google Scholar
  31. X. Wang, Y. Zhang, Z. Han, and K. He, “Malignancy of Cancers and synthetic lethal interactions associated with mutations of cancer driver genes,” Medicine, vol. 95, no. 8, p. e2697, 2016. View at: Publisher Site | Google Scholar
  32. K. Tanaka, M. Hashizume, M. Mihara, H. Yoshida, M. Suzuki, and Y. Matsumoto, “Anti-interleukin-6 receptor antibody prevents systemic bone mass loss via reducing the number of osteoclast precursors in bone marrow in a collagen-induced arthritis model,” Clinical & Experimental Immunology, vol. 175, no. 2, pp. 172–180, 2014. View at: Publisher Site | Google Scholar
  33. X. Luo, Y. Chen, G. Lv et al., “Adenovirus-mediated small interfering RNA targeting TAK1 ameliorates joint inflammation with collagen-induced arthritis in mice,” Inflammation, vol. 40, no. 3, pp. 894–903, 2017. View at: Publisher Site | Google Scholar
  34. R. S. Peres, G. B. Santos, N. T. Cecilio et al., “Lapachol, a compound targeting pyrimidine metabolism, ameliorates experimental autoimmune arthritis,” Arthritis Research & Therapy, vol. 19, no. 1, pp. 47–59, 2017. View at: Publisher Site | Google Scholar
  35. G. Lu, J. Liao, G. Yang, K. R. Reuhl, X. Hao, and C. S. Yang, “Inhibition of adenoma progression to adenocarcinoma in a 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced lung tumorigenesis model in A/J mice by tea polyphenols and caffeine,” Cancer Research, vol. 66, no. 23, pp. 11494–11501, 2006. View at: Publisher Site | Google Scholar
  36. A. Chatzikyriakidou, P. V. Voulgari, and A. A. Drosos, “Lack of association of the autophagy-related gene polymorphism ATG16L1 rs2241880 in RA predisposition,” Rheumatology International, vol. 34, no. 4, pp. 477–479, 2014. View at: Publisher Site | Google Scholar
  37. D. J. Klionsky, K. Abdelmohsen, A. Abe, M. J. Abedin et al., “Guidelines for the use and interpretation of assays for monitoring autophagy,” Autophagy, vol. 12, pp. 1–222, 2016. View at: Google Scholar
  38. K. K. Deeb, D. L. Trump, and C. S. Johnson, “Vitamin D signalling pathways in cancer: potential for anticancer therapeutics,” Nature Reviews Cancer, vol. 7, no. 9, pp. 684–700, 2007. View at: Publisher Site | Google Scholar
  39. K. W. Nettles, “Insights into PPAR gamma from structures with endogenous and covalently bound ligands,” Nature Structural & Molecular Biology, vol. 15, no. 9, pp. 893–895, 2008. View at: Publisher Site | Google Scholar
  40. F. Ge, Y. Zhang, F. Cao et al., “Arsenic trioxide-based therapy is suitable for patients with psoriasis-associated acute promyelocytic leukemia – A retrospective clinical study,” International Journal of Hematology, vol. 21, no. 5, pp. 287–294, 2016. View at: Publisher Site | Google Scholar
  41. T.-D. Zhang, G.-Q. Chen, Z.-G. Wang, Z.-Y. Wang, S.-J. Chen, and Z. Chen, “Arsenic trioxide, a therapeutic agent for APL,” Oncogene, vol. 20, no. 49, pp. 7146–7153, 2001. View at: Publisher Site | Google Scholar
  42. W. Zhang, Y. Liu, M. Ge et al., “Protective effect of resveratrol on arsenic trioxide-induced nephrotoxicity in rats,” Nutrition Research and Practice, vol. 8, no. 2, pp. 220–226, 2014. View at: Publisher Site | Google Scholar
  43. R. W. Ahn, S. L. Barrett, M. R. Raja et al., “Nano-encapsulation of arsenic trioxide enhances efficacy against murine lymphoma model while minimizing its impact on ovarian reserve in vitro and in vivo,” PLoS ONE, vol. 8, no. 3, Article ID e58491, 2013. View at: Google Scholar

Copyright © 2019 Weiyan Wang et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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