Duhuo Jisheng Decoction‑containing serum promotes proliferation of interleukin‑1β‑induced chondrocytes through the p16‑cyclin D1/CDK4‑Rb pathway

Wu,Guangwen; Fan,Huailing; Huang,Yuanpeng; Zheng,Chunsong; Ye,Jinxia; Liu,Xianxiang

doi:10.3892/mmr.2014.2527

Duhuo Jisheng Decoction‑containing serum promotes proliferation of interleukin‑1β‑induced chondrocytes through the p16‑cyclin D1/CDK4‑Rb pathway

Authors:
- Guangwen Wu
- Huailing Fan
- Yuanpeng Huang
- Chunsong Zheng
- Jinxia Ye
- Xianxiang Liu
View Affiliations

Affiliations: Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fujian, Fuzhou 350122, P.R. China, Academy of Traditional Chinese Medicine, Fujian University of Traditional Chinese Medicine, Fujian, Fuzhou 350122, P.R. China, Fujian Sports Vocational Education and Technical College, Fujian, Fuzhou 350003, P.R. China
Published online on: September 2, 2014 https://doi.org/10.3892/mmr.2014.2527
Pages: 2525-2534

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

Duhuo Jisheng Decoction (DHJSD) is a traditional Chinese herbal medicine that has multiple uses, including as a treatment for osteoarthritis (OA). However, the molecular mechanisms underlying the therapeutic effects of DHJSD on OA remain unknown. In the present study, a serum pharmacological method was applied to investigate the effects of DHJSD on the proliferation of chondrocytes treated with interleukin‑1β (IL‑1β) in vitro. This is a cell model commonly used to reproduce the mechanisms involved in degenerative arthropathies, including OA. The most effective intervention conditions of DHJSD serum were examined by MTT assay. The degenerative chondrocyte model was established by IL‑1β‑culture for 24 h, and was verified by optical microscopy and immunohistochemical analyses. Following the successful establishment of the degenerative chondrocyte model, the chondrocytes were subsequently randomly divided into two groups: The blank serum group and the DHJSD treatment group. Subsequent to treatment with the corresponding serum, cell proliferation was detected by MTT assay and DNA staining followed by FACS analysis, and the mRNA and protein expression levels of cyclin D1, cyclin‑dependent kinase 4 (CDK4), retinoblastoma tumor suppressor protein (Rb) and p16 were measured by reverse transcription polymerase chain reaction and western blotting, respectively. The results indicated that the most effective condition for the promotion of chondrocyte proliferation was 10% concentration of DHJSD 2‑h serum, and the degenerative chondrocyte model was successfully reproduced by IL‑1β‑treatment for 24 h. The mRNA and protein expression levels of cyclin D1, CDK4 and Rb in the DHJSD serum‑treated cells were significantly increased compared with those in the blank serum group, whereas p16 expression was significantly downregulated. These results indicate that treatment of cells with DHJSD‑containing serum is able to promote IL‑1β‑induced chondrocyte proliferation by promoting G1/S phase transition via modulating the expressions of cyclin D1, CDK4, Rb and p16, which contribute to the clinical efficacy of DHJSD in OA.

View Figures

View References

1	Felson DT: Clinical practice. Osteoarthritis of the knee. N Engl J Med. 354:841–848. 2006. View Article : Google Scholar : PubMed/NCBI
2	Zhang W, Nuki G, Moskowitz RW, et al: OARSI recommendations for the management of hip and knee osteoarthritis: part III: Changes in evidence following systematic cumulative update of research published through January 2009. Osteoarthritis Cartilage. 18:476–499. 2010. View Article : Google Scholar
3	Qiu G: Osteoarthritis diagnosis and treatment guidelines. Chinese J Joint Surgery. 1:281–285. 2007.
4	Shortkroff S and Yates KE: Alteration of matrix glycosaminoglycans diminishes articular chondrocytes’ response to a canonical Wnt signal. Osteoarthritis Cartilage. 15:147–154. 2007.PubMed/NCBI
5	Chan BY, Fuller ES, Russell AK, et al: Increased chondrocyte sclerostin may protect against cartilage degradation in osteoarthritis. Osteoarthritis Cartilage. 19:874–885. 2011. View Article : Google Scholar : PubMed/NCBI
6	Huang JG, Xia C, Zheng XP, et al: 17β-Estradiol promotes cell proliferation in rat osteoarthritis model chondrocytes via PI3K/Akt pathway. Cell Mol Biol Lett. 16:564–575. 2011.
7	Kashiwagi A, Schipani E, Fein MJ, Greer PA and Shimada M: Targeted deletion of Capn4 in cells of the chondrocyte lineage impairs chondrocyte proliferation and differentiation. Mol Cell Biol. 30:2799–2810. 2010. View Article : Google Scholar : PubMed/NCBI
8	Sarzi-Puttini P, Cimmino MA, Scarpa R, et al: Osteoarthritis: an overview of the disease and its treatment strategies. Semin Arthritis Rheum. 35(Suppl 1): 1–10. 2005. View Article : Google Scholar
9	Badley EM, Rasooly I and Webster GK: Relative importance of musculoskeletal disorders as a cause of chronic health problems, disability, and health care utilization: findings from the 1990 Ontario Health Survey. J Rheumatol. 21:505–514. 1994.
10	Felson DT, Lawrence RC, Dieppe PA, et al: Osteoarthritis: new insights. Part 1: the disease and its risk factors. Ann Intern Med. 133:635–646. 2000. View Article : Google Scholar : PubMed/NCBI
11	Bradley JD, Brandt KD, Katz BP, Kalasinski LA and Ryan SI: Comparison of an antiinflammatory dose of ibuprofen, an analgesic dose of ibuprofen, and acetaminophen in the treatment of patients with osteoarthritis of the knee. N Engl J Med. 325:87–91. 1991. View Article : Google Scholar : PubMed/NCBI
12	Lohmander LS, Dalén N, Englund G, et al: Intra-articular hyaluronan injections in the treatment of osteoarthritis of the knee: a randomised, double blind, placebo controlled multicentre trial. Ann Rheum Dis. 55:424–431. 1996. View Article : Google Scholar : PubMed/NCBI
13	Berenbaum F: New horizons and perspectives in the treatment of osteoarthritis. Arthritis Res Ther. 10(Suppl 2): S12008. View Article : Google Scholar : PubMed/NCBI
14	Ahmed S, Anuntiyo J, Malemud CJ and Haqqi TM: Biological basis for the use of botanicals in osteoarthritis and rheumatoid arthritis: a review. Evid Based Complement Alternat Med. 2:301–308. 2005. View Article : Google Scholar : PubMed/NCBI
15	Parkman CA: Alternative therapies for osteoarthritis. Case Manager. 12:34–36. 2001. View Article : Google Scholar
16	Teekachunhatean S, Kunanusorn P, Rojanasthien N, et al: Chinese herbal recipe versus diclofenac in symptomatic treatment of osteoarthritis of the knee: a randomized controlled trial [ISRCTN70292892]. BMC Complement Altern Med. 4:192004.PubMed/NCBI
17	Setty AR and Sigal LH: Herbal medications commonly used in the practice of rheumatology: mechanisms of action, efficacy, and side effects. Semin Arthritis Rheum. 34:773–784. 2005. View Article : Google Scholar : PubMed/NCBI
18	Chun SC, Jee SY, Lee SG, Park SJ, Lee JR and Kim SC: Anti-inflammatory activity of the methanol extract of moutan cortex in LPS-activated Raw264.7 cells. Evid Based Complement Alternat Med. 4:327–333. 2007. View Article : Google Scholar : PubMed/NCBI
19	Wu M and Gu Z: Screening of bioactive compounds from moutan cortex and their anti-inflammatory activities in rat synoviocytes. Evid Based Complement Alternat Med. 6:57–63. 2009. View Article : Google Scholar : PubMed/NCBI
20	Sun SM: Bei Ji Qian Jin Yao Fang. 8. 1st edition. People’s Medical Publishing Press; Beijing: pp. 166–167. 1982
21	Lai JN, Chen HJ, Chen CC, Lin JH, Hwang JS and Wang JD: Duhuo jisheng tang for treating osteoarthritis of the knee: a prospective clinical observation. Chin Med. 2:42007. View Article : Google Scholar : PubMed/NCBI
22	Zheng CS, XXJ, Ye HZ, Wu GW, Li XH, Huang SP and Liu XX: Computational approaches for exploring the potential synergy and polypharmacology of Duhuo Jisheng Decoction in the therapy of osteoarthritis. Mol Med Report. 7:1812–1818. 2013.PubMed/NCBI
23	Wu G, Chen W, Fan H, et al: Duhuo Jisheng Decoction promotes chondrocyte proliferation through accelerated G1/S transition in osteoarthritis. Int J Mol Med. 32:1001–1010. 2013.PubMed/NCBI
24	Panico AM, Cardile V, Garufi F, Puglia C, Bonina F and Ronsisvalle G: Effect of hyaluronic acid and polysaccharides from Opuntia ficus indica (L.) cladodes on the metabolism of human chondrocyte cultures. J Ethnopharmacol. 111:315–321. 2007.PubMed/NCBI
25	The Ministry of Science and Technology of the People’s Republic of China. Guidance Suggestions for the Care and Use of Laboratory Animals. 2006.
26	Huang J, Huang X, Chen Z, Zheng Q and Sun R: Dose conversion among different animals and healthy volunteers in pharmacological study. Chin J Clin Pharmacol Ther. 9:1069–1072. 2004.(In Chinese).
27	Li XH, Du M, Liu XX, et al: Millimeter wave treatment inhibits NO-induced apoptosis of chondrocytes through the p38MAPK pathway. Int J Mol Med. 25:393–399. 2010.PubMed/NCBI
28	Li XH, Wu MX, YHZ, Chen WL, Lin JM, Zheng LP and Liu XX: Experimental study on the suppression of sodium nitroprussiate-induced chondrocyte apoptosis by tougu xiaotong capsule-containing serum. Chin J Integr Med. 17:436–443. 2011.(In Chinese).
29	Sanchez C, Mathy-Hartert M, Deberg MA, Ficheux H, Reginster JY and Henrotin YE: Effects of rhein on human articular chondrocytes in alginate beads. Biochem Pharmacol. 65:377–388. 2003. View Article : Google Scholar : PubMed/NCBI
30	Wu SQ, Otero M, Unger FM, et al: Anti-inflammatory activity of an ethanolic Caesalpinia sappan extract in human chondrocytes and macrophages. J Ethnopharmacol. 138:364–372. 2011. View Article : Google Scholar : PubMed/NCBI
31	Mankin HJ: The response for articular cartilage to mechanical injury. J Bone Joint Surg Am. 64:460–466. 1982.PubMed/NCBI
32	Brandt KD and Mazzuca SA: Lessons learned from nine clinical trials of disease-modifying osteoarthritis drugs. Arthritis Rheum. 52:3349–3359. 2005. View Article : Google Scholar : PubMed/NCBI
33	Bensky D, Gamble A and Stöger E: Chinese Herbal Medicine: Materia Medica. 3rd edition. Eastland Press; Seattle, WA: 2004
34	Ho LJ and Lai JH: Chinese herbs as immunomodulators and potential disease-modifying antirheumatic drugs in autoimmune disorders. Curr Drug Metab. 5:181–192. 2004. View Article : Google Scholar : PubMed/NCBI
35	Tashino S: ‘Serum pharmacology’ and ‘serum pharmaceutical chemistry’: from pharmacology of Chinese traditional medicines to start a new measurement of drug concentration in blood. Ther Drug Monit Res. 5:54–64. 1988.
36	Liu N, Liu JT, Ji YY and Lu PP: Dahuang zhechong pill containing serum inhibited platelet-derived growth factor-stimulated vascular smooth muscle cells proliferation by inducing G1 arrest partly via suppressing protein kinase C α-extracellular regulated kinase 1/2 signaling. Chin J Integr Med. 18:371–377. 2012.(In Chinese).
37	Dang XY, Dong L, Shi HT and Zou BC: Effects of serum containing Chinese medicine Sanpi Pingwei formula on proliferation and apoptosis of human SGC-7901 cells. Chin J Integr Med. 19:119–126. 2013. View Article : Google Scholar : PubMed/NCBI
38	Wang WJ, Li DJ, Li J and Zhou WJ: An in vitro study on neuroprotective effects of serum containing Gengnianchun decoction and its main monomers against amyloid beta protein-induced cellular toxicity. J Chin Integr Med. 8:67–73. 2010.(In Chinese).
39	Henrotin Y, Sanchez C and Reginster JY: The inhibition of metalloproteinases to treat osteoarthritis: reality and new perspectives. Expert Opin Ther Patents. 12:29–43. 2002. View Article : Google Scholar
40	Tetlow LC, Adlam DJ and Woolley DE: Matrix metalloproteinase and proinflammatory cytokine production by chondrocytes of human osteoarthritic cartilage. Arthritis Rheum. 44:585–594. 2001. View Article : Google Scholar : PubMed/NCBI
41	Martel-Pelletier J, Di Battista J and Lajeunesse D: Biochemical factors in joint articular tissue degradation in osteoarthritis. Osteoarthritis: Clinical and Experimental Aspects. Reginster J-YL, Pelletier JP, Martel-Pelletier J and Henrotin YE: Springer; Berlin: pp. 156–187. 1999, View Article : Google Scholar
42	Park K, Hoffmeister B, Han DK and Hasty K: Therapeutic ultrasound effects on interleukin-1beta stimulated cartilage construct in vitro. Ultrasound Med Biol. 33:286–295. 2007. View Article : Google Scholar : PubMed/NCBI
43	Tan L, Peng H, Osaki M, Choy BK, Auron PE, Sandell LJ and Goldring MB: Egr-1 mediates transcriptional repression of COL2A1 promoter activity by interleukin-1beta. J Biol Chem. 278:17688–17700. 2003. View Article : Google Scholar : PubMed/NCBI
44	Gibson SL, Dai CY, Lee HW, et al: Inhibition of colon tumor progression and angiogenesis by the Ink4a/Arf locus. Cancer Res. 63:742–746. 2003.PubMed/NCBI
45	Massagué J: G1 cell-cycle control and cancer. Nature. 432:298–306. 2004.
46	Musgrove EA, Caldon CE, Barraclough J, Stone A and Sutherland RL: Cyclin D as a therapeutic target in cancer. Nat Rev Cancer. 11:558–572. 2011. View Article : Google Scholar : PubMed/NCBI
47	Li J, Poi MJ and Tsai MD: Regulatory mechanisms of tumor suppressor P16(INK4A) and their relevance to cancer. Biochemistry. 50:5566–5582. 2011. View Article : Google Scholar : PubMed/NCBI
48	Witkiewicz AK, Knudsen KE, Dicker AP and Knudsen ES: The meaning of p16(ink4a) expression in tumors: functional significance, clinical associations and future developments. Cell Cycle. 10:2497–2503. 2011. View Article : Google Scholar : PubMed/NCBI