Pro‑neurogenic effects of andrographolide on RSC96 Schwann cells in vitro

Xu,Fuben; Wu,Huayu; Zhang,Kun; Lv,Peizhen; Zheng,Li; Zhao,Jinmin

doi:10.3892/mmr.2016.5717

Pro‑neurogenic effects of andrographolide on RSC96 Schwann cells in vitro

Authors:
- Fuben Xu
- Huayu Wu
- Kun Zhang
- Peizhen Lv
- Li Zheng
- Jinmin Zhao
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Affiliations: Research Center for Regenerative Medicine, Guangxi Medical University, Nanning, Guangxi 530021, P.R. China, Department of Cell Biology and Genetics, School of Premedical Sciences, Guangxi Medical University, Nanning, Guangxi 530021, P.R. China
Published online on: September 6, 2016 https://doi.org/10.3892/mmr.2016.5717
Pages: 3573-3580
Copyright: © Xu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Nerve regeneration remains a challenge to the treatment of peripheral nerve injury. Andrographolide (Andro) is the main active constituent of Andrographis paniculata, which has been applied in the treatment of several diseases, including inflammation, in ancient China. Andro has been reported to facilitate the reduction of edema and to exert analgesic effects in the treatment of various diseases. These findings suggest that Andro may be considered a promising anti‑inflammatory agent that may suppress destruction and accelerate proliferation of Schwann cells following peripheral nerve injury. In the present study, the effects of Andro on RSC96 cells were investigated in vitro. The RSC96 cell line is a spontaneously immortalized rat Schwann cell line, which was originally derived from a long‑term culture of rat primary Schwann cells. RSC96 cells were treated with a range of 0 to 50 µM Andro prior to the MTT assay. Cell proliferation, morphology, synthesis and nerve‑specific gene expression were performed to detect the effect of Andro on RSC96 cells. The results of the present study demonstrated that the recommended doses of Andro ranged between 0.78 and 12.5 µM, among which the most obvious response was observed when used at 3.125 µM (P<0.05). DNA content was improved in Andro groups compared with the control group (P<0.05). In addition, Andro was able to promote the gene expression of glial cell line‑derived neurotrophic factor, brain‑derived neurotrophic factor, ciliary neurotrophic factor, and the specific Schwann cell marker S100β (P<0.05). The results of a viability assay, hematoxylin‑eosin staining, and immunohistochemistry were also improved in Andro groups. These results indicated that Andro may accelerate proliferation of RSC96 cells in vitro, whilst maintaining the Schwann cell phenotype; therefore, the present study may provide valuable evidence for the further exploration of the effects of Andro on peripheral nerves.

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1	Lundborg G: A 25-year perspective of peripheral nerve surgery: Evolving neuroscientific concepts and clinical significance. J Hand Surg Am. 25:391–414. 2000. View Article : Google Scholar : PubMed/NCBI
2	Robinson LR: Traumatic injury to peripheral nerves. Suppl Clin Neurophysiol. 57:173–186. 2004. View Article : Google Scholar
3	Evans GR: Peripheral nerve injury: A review and approach to tissue engineered constructs. Anat Rec. 263:396–404. 2001. View Article : Google Scholar : PubMed/NCBI
4	Raine CS: Multiple sclerosis: Immune system molecule expression in the central nervous system. J Neuropathol Exp Neurol. 53:328–337. 1994. View Article : Google Scholar : PubMed/NCBI
5	Rogers J and Shen Y: A perspective on inflammation in Alzheimer's disease. Ann N Y Acad Sci. 924:132–135. 2000. View Article : Google Scholar
6	Qin L, Liu Y, Wang T, Wei SJ, Block ML, Wilson B, Liu B and Hong JS: NADPH oxidase mediates lipopolysaccharide-induced neurotoxicity and proinflammatory gene expression in activated microglia. J Biol Chem. 279:1415–1421. 2004. View Article : Google Scholar
7	Zochodne DW: The microenvironment of injured and regenerating peripheral nerves. Muscle Nerve Suppl. 9:S33–S38. 2000. View Article : Google Scholar
8	Dezawa M: Central and peripheral nerve regeneration by transplantation of Schwann cells and transdifferentiated bone marrow stromal cells. Anat Sci Int. 77:12–25. 2002. View Article : Google Scholar : PubMed/NCBI
9	Wang L, Sanford MT, Xin Z, Lin G and Lue TF: Role of Schwann cells in the regeneration of penile and peripheral nerves. Asian J Androl. 17:776–782. 2015.PubMed/NCBI
10	Puri A, Saxena R, Saxena RP, Saxena KC, Srivastava V and Tandon JS: Immunostimulant agents from Andrographis paniculata. J Nat Prod. 56:995–999. 1993. View Article : Google Scholar : PubMed/NCBI
11	Zhang XF and Tan BK: Antihyperglycaemic and anti-oxidant properties of Andrographis paniculata in normal and diabetic rats. Clin Exp Pharmacol Physiol. 27:358–363. 2000. View Article : Google Scholar : PubMed/NCBI
12	Shen YC, Chen CF and Chiou WF: Andrographolide prevents oxygen radical production by human neutrophils: Possible mechanism(s) involved in its anti-inflammatory effect. Br J Pharmacol. 135:399–406. 2002. View Article : Google Scholar : PubMed/NCBI
13	Zhu T, Wang DX, Zhang W, Liao XQ, Guan X, Bo H, Sun JY, Huang NW, He J, Zhang YK, et al: Andrographolide protects against LPS-induced acute lung injury by inactivation of NF-κB. PLoS One. 8:e564072013. View Article : Google Scholar
14	Bao Z, Guan S, Cheng C, Wu S, Wong SH, Kemeny DM, Leung BP and Wong WS: A novel antiinflammatory role for andrographolide in asthma via inhibition of the nuclear factor-kappaB pathway. Am J Respir Crit Care Med. 179:657–665. 2009. View Article : Google Scholar : PubMed/NCBI
15	Chen JX, Xue HJ, Ye WC, Fang BH, Liu YH, Yuan SH, Yu P and Wang YQ: Activity of andrographolide and its derivatives against influenza virus in vivo and in vitro. Biol Pharm Bull. 32:1385–1391. 2009. View Article : Google Scholar : PubMed/NCBI
16	Li J, Luo L, Wang X, Liao B and Li G: Inhibition of NF-kappaB expression and allergen-induced airway inflammation in a mouse allergic asthma model by andrographolide. Cell Mol Immunol. 6:381–385. 2009. View Article : Google Scholar : PubMed/NCBI
17	Wang T, Liu B, Zhang W, Wilson B and Hong JS: Andrographolide reduces inflammation-mediated dopaminergic neurodegeneration in mesencephalic neuron-glia cultures by inhibiting microglial activation. J Pharmacol Exp Ther. 308:975–983. 2004. View Article : Google Scholar : PubMed/NCBI
18	Wen L, Xia N, Chen X, Li Y, Hong Y and Liu Y, Wang Z and Liu Y: Activity of antibacterial, antiviral, anti-inflammatory in compounds andrographolide salt. Eur J Pharmacol. 740:421–427. 2014. View Article : Google Scholar : PubMed/NCBI
19	Ku CM and Lin JY: Anti-inflammatory effects of 27 selected terpenoid compounds tested through modulating Th1/Th2 cytokine secretion profiles using murine primary splenocytes. Food Chem. 141:1104–1113. 2013. View Article : Google Scholar : PubMed/NCBI
20	Chen YY, Hsu MJ, Sheu JR, Lee LW and Hsieh CY: Andrographolide, a novel NF- κB inhibitor, induces vascular smooth muscle cell apoptosis via a Ceramide-p47phox-ROS signaling cascade. Evid Based Complement Alternat Med. 2013:8218132013.
21	Talei D, Valdiani A, Maziah M, Sagineedu SR and Saad MS: Analysis of the anticancer phytochemicals in andrographis paniculata Nees. Under salinity stress. Biomed Res Int. 2013:3190472013. View Article : Google Scholar : PubMed/NCBI
22	Manikam SD and Stanslas J: Andrographolide inhibits growth of acute promyelocytic leukaemia cells by inducing retinoic acid receptor-independent cell differentiation and apoptosis. J Pharm Pharmacol. 61:69–78. 2009. View Article : Google Scholar : PubMed/NCBI
23	Lehmann HC and Höke A: Schwann cells as a therapeutic target for peripheral neuropathies. CNS Neurol Disord Drug Targets. 9:801–806. 2010. View Article : Google Scholar : PubMed/NCBI
24	Huang J, Hu X, Lu L, Ye Z, Zhang Q and Luo Z: Electrical regulation of Schwann cells using conductive polypyrrole/chitosan polymers. J Biomed Mater Res A. 93:164–174. 2010.
25	Rotshenker S: Wallerian degeneration: The innate-immune response to traumatic nerve injury. J Neuroinflammation. 8:1092011. View Article : Google Scholar : PubMed/NCBI
26	Bosse F: Extrinsic cellular and molecular mediators of peripheral axonal regeneration. Cell Tissue Res. 349:5–14. 2012. View Article : Google Scholar : PubMed/NCBI
27	Yuan H, Zhang J, Liu H and Li Z: The protective effects of resveratrol on Schwann cells with toxicity induced by ethanol in vitro. Neurochem Int. 63:146–153. 2013. View Article : Google Scholar : PubMed/NCBI
28	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
29	Chan SJ, Wong WS, Wong PT and Bian JS: Neuroprotective effects of andrographolide in a rat model of permanent cerebral ischaemia. Br J Pharmacol. 161:668–679. 2010. View Article : Google Scholar : PubMed/NCBI
30	Zaitone SA, Abo-Elmatty DM and Shaalan AA: Acetyl-L-carnitine and α-lipoic acid affect rotenone-induced damage in nigral dopaminergic neurons of rat brain, implication for Parkinson's disease therapy. Pharmacol Biochem Behav. 100:347–360. 2012. View Article : Google Scholar
31	Jalali-Nadoushan M and Roghani M: Alpha-lipoic acid protects against 6-hydroxydopamine-induced neurotoxicity in a rat model of hemi-parkinsonism. Brain Res. 1505:68–74. 2013. View Article : Google Scholar : PubMed/NCBI
32	Zhang Z, Lai D, Wang L, Yu P, Zhu L, Guo B, Xu L, Zhou L, Sun Y, Lee SM and Wang Y: Neuroprotective effects of the andrographolide analogue AL-1 in the MPP⁺/MPTP-induced Parkinson's disease model in vitro and in mice. Pharmacol Biochem Behav. 122:191–202. 2014. View Article : Google Scholar : PubMed/NCBI
33	Aloe L, Rocco ML, Bianchi P and Manni L: Nerve growth factor: From the early discoveries to the potential clinical use. J Transl Med. 10:2392012. View Article : Google Scholar : PubMed/NCBI
34	Bothwell M: NGF, BDNF, NT3, and NT4. Handb Exp Pharmacol. 220:3–15. 2014. View Article : Google Scholar : PubMed/NCBI
35	Daly W, Yao L, Zeugolis D, Windebank A and Pandit A: A biomaterials approach to peripheral nerve regeneration: Bridging the peripheral nerve gap and enhancing functional recovery. J R Soc Interface. 9:202–221. 2012. View Article : Google Scholar
36	Klimaschewski L, Hausott B and Angelov DN: The pros and cons of growth factors and cytokines in peripheral axon regeneration. Int Rev Neurobiol. 108:137–171. 2013. View Article : Google Scholar : PubMed/NCBI
37	Xie Y and Longo FM: Neurotrophin small-molecule mimetics. Prog Brain Res. 128:333–347. 2000. View Article : Google Scholar : PubMed/NCBI
38	Peleshok J and Saragovi HU: Functional mimetics of neurotrophins and their receptors. Biochem Soc Trans. 34:612–617. 2006. View Article : Google Scholar : PubMed/NCBI
39	Colangelo AM, Bianco MR, Vitagliano L, Cavaliere C, Cirillo G, De Gioia L, Diana D, Colombo D, Redaelli C, Zaccaro L, et al: A new nerve growth factor-mimetic peptide active on neuropathic pain in rats. J Neurosci. 28:2698–2709. 2008. View Article : Google Scholar : PubMed/NCBI
40	Gordon T: The role of neurotrophic factors in nerve regeneration. Neurosurg Focus. 26:E32009. View Article : Google Scholar : PubMed/NCBI
41	Zhu H, Wang WJ, Ding WL, Li F and He J: Effect of panaxydol on hypoxia-induced cell death and expression and secretion of neurotrophic factors (NTFs) in hypoxic primary cultured Schwann cells. Chem Biol Interact. 174:44–50. 2008. View Article : Google Scholar : PubMed/NCBI
42	Donato R: S100: A multigenic family of calcium-modulated proteins of the EF-hand type with intracellular and extracellular functional roles. Int J Biochem Cell Biol. 33:637–668. 2001. View Article : Google Scholar : PubMed/NCBI
43	Donato R: Intracellular and extracellular roles of S100 proteins. Microsc Res Tech. 60:540–551. 2003. View Article : Google Scholar : PubMed/NCBI
44	Novitskaya V, Grigorian M, Kriajevska M, Tarabykina S, Bronstein I, Berezin V, Bock E and Lukanidin E: Oligomeric forms of the metastasis-related Mts1 (S100A4) protein stimulate neuronal differentiation in cultures of rat hippocampal neurons. J Biol Chem. 275:41278–41286. 2000. View Article : Google Scholar : PubMed/NCBI