Protective effect of pyrroloquinoline quinine on ultraviolet A irradiation‑induced human dermal fibroblast senescence in vitro proceeds via the anti‑apoptotic sirtuin 1/nuclear factor‑derived erythroid 2‑related factor 2/heme oxygenase 1 pathway

Zhang,Chunli; Wen,Chuanjun; Lin,Jinde; Shen,Gan

doi:10.3892/mmr.2015.3990

Protective effect of pyrroloquinoline quinine on ultraviolet A irradiation‑induced human dermal fibroblast senescence in vitro proceeds via the anti‑apoptotic sirtuin 1/nuclear factor‑derived erythroid 2‑related factor 2/heme oxygenase 1 pathway

Authors:
- Chunli Zhang
- Chuanjun Wen
- Jinde Lin
- Gan Shen
View Affiliations

Affiliations: Department of Clinical Research, Friendship Plastic Surgery Hospital, Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China, College of Science, Nanjing Normal University, Nanjing, Jiangsu 210046, P.R. China, Department of Plastic Surgery, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210000, P.R. China
Published online on: June 24, 2015 https://doi.org/10.3892/mmr.2015.3990
Pages: 4382-4388

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

The aim of the present study was to determine whether pyrroloquinoline quinine (PQQ) exerts a protective effect on ultraviolet A (UVA) irradiation‑induced senescence in human dermal fibroblasts (HDFs) and to elucidate its mechanism of action in vitro. A senescence model was constructed as follows: HDFs (1x104‑1x106) were cultured in a six‑well plate in vitro and then exposed to UVA irradiation at a dosage of 9 J/cm2. Various concentrations of PQQ (50, 100 and 200 ng/ml) were added to the culture medium 24 h prior to UVA exposure. Following 72 h of irradiation, senescence‑associated β‑galactosidase staining was performed in order to evaluate the senescence state. Furthermore, mRNA expression of the senescence marker genes matrix‑metalloprotease (MMP)1 and MMP3 was determined using reverse transcription quantitative polymerase chain reaction. Protein expression of sirtuin (SIRT)1, SIRT6, nuclear factor erythroid 2‑related factor 2 (Nrf2) and heme oxygenase 1 (HO‑1) were detected using western blot analysis. The results showed that the percentage of cells stained by X‑gal following 9 J/cm2 UVA irradiation was markedly increased compared with that of the control group (53 and 8%, respectively), while 50 ng/ml PQQ attenuated the ratio of positive staining compared with that of the UVA‑only cells (29 vs. 53%, respectively). Expression of fibroblast senescence marker genes MMP1 and MMP3 was decreased in cells treated with UVA and 50 ng/ml PQQ compared with that of cells in the UVA‑only group. Western blot analysis revealed significant effects of PQQ on SIRT1 and SIRT6. Nrf2 and HO‑1 exbibited mild changes with the same trend when treated with or without UVA and PQQ. In conclusion, the results of the present study showed that pyrroloquinoline quinine may have a protective effect on UVA irradiation‑induced HDF aging, which may be associated with the anti‑apoptotic SIRT1/Nrf2/HO‑1 pathway as well as SIRT6 signaling.

View Figures

View References

1	Kohl E, Steinbauer J, Landthaler M and Szeimies RM: Skin aging. J Eur Acad Dermatol. 25:873–884. 2011. View Article : Google Scholar
2	He YY, Council SE, Feng L and Chignell CF: UVA-induced cell cycle progression is mediated by a disintegrin and metal-loprotease/epidermal growth factor receptor/AKT/Cyclin D1 pathways in keratinocytes. Cancer Res. 68:3752–3758. 2008. View Article : Google Scholar : PubMed/NCBI
3	Krutmann J: Ultraviolet A radiation-induced biological effects in human skin: relevance for photo aging and photodermatosis. J Dermatol Sci. 23(Suppl 1): S22–S26. 2000. View Article : Google Scholar
4	Salisbury SA, Forrest HS, Cruse WB and Kennard O: A novel coenzyme from bacterial primary alcohol dehydrogenases. Nature. 280:843–844. 1979. View Article : Google Scholar : PubMed/NCBI
5	Felton LM and Anthony C: Biochemistry: role of PQQ as a mammalian enzyme cofactor? Nature. 433:E10–E12. 2005. View Article : Google Scholar : PubMed/NCBI
6	Matsushitak K, Toyama H, Yamada M, et al: Quinoproteins: structure function and biotechnologies applications. Appl Microboil Biot. 58:13–22. 2002. View Article : Google Scholar
7	Liu CL, Siesjö BK and Hu BR: Pathogenesis of hippocampal neuronal death after hypoxia-ischemia changes during brain development. Neuroscience. 127:113–123. 2004. View Article : Google Scholar : PubMed/NCBI
8	Dimri GP, Lee X, Basile G, et al: A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Cell Biology. 92:9363–9367. 1995.
9	Ohwada K, Takeda H, Yamazaki M, Isogai H, Nakano M, Shimomura M, Fukui K and Urano S: Pyrroloquinoline quinone (PQQ) prevents cognitive deficit caused by oxidative stress in rats. J Clin Biochem Nutr. 42:29–34. 2008. View Article : Google Scholar : PubMed/NCBI
10	Zhang Q, Shen M, Ding M, Shen D and Ding F: The neuropro-tective action of pyrroloquinoline against glutamate-induced apoptosis in hippocampal neurons is mediated through the activation of PI3K/Akt pathway. Toxicol Appl Pharm. 252:62–72. 2011. View Article : Google Scholar
11	Kanfi Y, Naiman S, Amir G, et al: The sirtuin SIRT6 regulates lifespan in male mice. Nature. 483:218–221. 2012. View Article : Google Scholar : PubMed/NCBI
12	Hekimi S and Guarente L: Genetics and the specificity of the aging process. Science. 299:1351–1354. 2003. View Article : Google Scholar : PubMed/NCBI
13	Howitz KT, Bitterman KJ, Cohen HY, et al: Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan. Nature. 425:191–196. 2003. View Article : Google Scholar : PubMed/NCBI
14	Landry J, Sutton A, Tafrov ST, et al: The silencing protein SIR2 and its homologs are NAD-dependent protein deacetylases. Proc Natl Acad Sci USA. 97:5807–5811. 2000. View Article : Google Scholar : PubMed/NCBI
15	Sugino T, Maruyama M, Tanno M, et al: Protein deacetylase SIRT1 in the cytoplasm promotes nerve growth factor-induced neurite outgrowth in PC12 cells. FEBS Lett. 584:2821–2826. 2010. View Article : Google Scholar : PubMed/NCBI
16	Hisahara S, Chiba S, Matsumoto H, et al: Histone deacetylase SIRT1 modulates neuronal differentiation by its nuclear translocation. Proc Natl Acad Sci USA. 105:15599–15604. 2008. View Article : Google Scholar : PubMed/NCBI
17	Michán S, Li Y, Chou MM, et al: SIRT1 is essential for normal cognitive function and synaptic plasticity. J Neurosci. 30:9695–9707. 2010. View Article : Google Scholar : PubMed/NCBI
18	Jin Q, Yan T, Ge X, Sun C, Shi X and Zhai Q: Cytoplasm-localized SIRT1 enhances apoptosis. J Cell Physiol. 213:88–97. 2007. View Article : Google Scholar : PubMed/NCBI
19	Byles V, Chmilewski LK, Wang J, et al: Aberrant cytoplasm localization and protein stability of SIRT1 is regulated by PI3K/IGF-1R signaling in human cancer cells. Int J Biol Sci. 6:599–612. 2010. View Article : Google Scholar : PubMed/NCBI
20	Herskovits AZ and Guarente L: Sirtuin deacelylase in neurode-generative disease of aging. Cell Res. 23:746–758. 2013. View Article : Google Scholar : PubMed/NCBI
21	Herrmann G, Brenneisen P, Wlaschek M, et al: Psoralen photo-activation promotes morphological and functional changes in fibroblasts in vitro reminiscent of cellular senescence. J Cell Sci. 111:759–767. 1998.
22	Brugè F, Venditti E, Tiano L, Littarru GP and Damiani E: Reference gene validation for qPCR on normoxia- and hypoxia-cultured human dermal fibroblasts exposed to UVA: is β-actin a reliable normalize for photoaging studies? J Biotechnol. 156:153–162. 2011. View Article : Google Scholar
23	Kawahara TL, Michishita E, Adler AS, et al: SIRT6 links histone H3 lysine 9 deacetylation and organismal life span. Cell. 136:62–74. 2008. View Article : Google Scholar
24	Herrmann G, Wlaschek M, Lange TS, et al: UVA irradiation stimulates the synthesis of various matrix-metalloproteinases (MMPs) in cultured human fibroblasts. Exp Dermatol. 2:92–97. 1993. View Article : Google Scholar : PubMed/NCBI
25	Masek T, Vopalensky V, Suchomelova P and Pospisek M: Denaturing RNA electrophoresis in TAE agrose gels. Anal Biochem. 336:46–50. 2005. View Article : Google Scholar
26	Vandesompele J, De Preter K, Pattyn F, et al: Accurate normalization of real-time quantitative RT-PCR data by geometric average of multiple internal control genes. Genome Biol. 3:RESEARCH00342002. View Article : Google Scholar
27	Li H, Wu S, Shi N, Lian S and Lin W: Nrf2/HO-1 pathway activation by manganese is associated with reactive oxygen species and ubiquitin-proteasome pathway, not MAPKs signaling. J Appl Toxicol. 31:690–697. 2011. View Article : Google Scholar : PubMed/NCBI
28	Kimura K, Takada M, Ishii T, et al: Pyrroloquinoline quinine stimulates epithelial cell proliferation by activating epidermal growth factor receptor through redox cycling. Free Radical Bio Med. 53:1239–1251. 2012. View Article : Google Scholar
29	Zhang Q, Ding M, Cao Z, et al: Pyrroloquinoline quinine protects rat brain cortex against acute glutamate-induced neurotoxicity. Neutovhrm Res. 38:1661–1671. 2013.
30	Li HH, He B, Peng H and Liu SQ: Effects of pyrroloquinoline quinine on proliferation and expression of c-fos, c-jun, CREB and PCNA in cultured Schwann cells. Zhonghua Zheng Xing Wai Ke Za Zhi. 27:298–303. 2011.In Chinese. PubMed/NCBI
31	Blander G and Guarente L: The Sir family of protein deaceylases. Annu Rev Biochem. 73:417–435. 2004. View Article : Google Scholar
32	Lin SJ, Defossez PA and Guarente L: Requirement of NAD and SIR2 for life-span extension by calorie restriction in Sacharomyces cerevisiae. Science. 289:2126–2128. 2000. View Article : Google Scholar : PubMed/NCBI
33	Frye RA: Characterization of five human cDNAs with homology to the yeast SIR2 gene: Sir2-like proteins (sirtuins) metabolize NAD and may have protein ADP-ribosyltransferase activity. Biochem Biophys Res Commun. 260:273–279. 1999. View Article : Google Scholar : PubMed/NCBI