Effects of ginsenoside Rb1 on oxidative stress injury in rat spinal cords by regulating the eNOS/Nrf2/HO‑1 signaling pathway

Liu,Xinwei; Gu,Xiaochuan; Yu,Miaomiao; Zi,Ying; Yu,Hailong; Wang,Yu; Xie,Yanchun; Xiang,Liangbi

doi:10.3892/etm.2018.6286

Effects of ginsenoside Rb1 on oxidative stress injury in rat spinal cords by regulating the eNOS/Nrf2/HO‑1 signaling pathway

Authors:
- Xinwei Liu
- Xiaochuan Gu
- Miaomiao Yu
- Ying Zi
- Hailong Yu
- Yu Wang
- Yanchun Xie
- Liangbi Xiang
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Affiliations: Department of Orthopedics, Rescue Center of Severe Wound and Trauma of the PLA, General Hospital of Shenyang Military Area Command, Shenyang, Liaoning 110016, P.R. China, Department of Orthopedics, Changhai Hospital Αffiliated to The Second Military Medical University, Shanghai 200433, P.R. China, Department of Emergency, Hospital 463 of the PLA, Shenyang, Liaoning 110042, P.R. China
Published online on: June 12, 2018 https://doi.org/10.3892/etm.2018.6286
Pages: 1079-1086
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study aimed to investigate whether ginsenoside Rb1 (G‑Rb1) attenuates spinal cord injury‑associated oxidative stress in rats by regulating the endothelial nitric oxide synthase eNOS/nuclear factor erythroid 2‑related factor 2 (Nrf2)/heme oxygenase (HO)‑1 signaling pathway. Sprague Dawley rats were randomly divided into the sham operation group (S group), spinal cord injury group (SCI group), G‑Rb1 treatment group (G‑Rb1 group) and SCI+G‑Rb1+Inhibitor L‑name group (L‑name group). The posterior limb function was evaluated via the Basso, Beattie and Bresnahan scoring method. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT) and glutathione (GSH) in serum were measured by ELISA. The pathological changes in the spinal cord were observed by H&E staining. Reverse transcription‑quantitative polymerase chain reaction and western blot analyses were used to detect eNOS, phosphorylated (p)‑eNOS, heat shock protein (HSP)90, Nrf2 and NAD(P)H quinone dehydrogenase 1 (Nqo1) at the mRNA and protein level. Immunohistochemistry was used to detect the expression of Nrf2 and p‑eNOS. Compared with the S group, the scores of spinal cord function in the SCI group were significantly lower, and the levels of MDA were significantly increased, while the levels of SOD, CAT and GSH protein in spinal cord were significantly decreased (P<0.05). The spinal cord tissue exhibited hemorrhage, neuronal degeneration/necrosis, as well as mononuclear cell and lymphocyte infiltration. The eNOS, HSP90, Nrf2, Nqo1 and HO‑1 mRNA levels were decreased (P<0.05). Compared with those in the SCI group, the spinal cord function score in the G‑Rb1 group were significantly higher and the serum MDA content was significantly decreased, while the activity of SOD, CAT and GSH was significantly increased (P<0.05). The degeneration/necrosis of spinal cord neurons was attenuated, inflammatory cell infiltration was significantly reduced and the levels of eNOS, HSP90, Nrf2, Nqo1 and HO‑1 were significantly upregulated (P<0.05). In the group that was administered the eNOS inhibitor L‑name, the levels of eNOS, HSP90, Nrf2, Nqo1 and HO‑1 were significantly decreased. In conclusion, G‑Rb1 attenuates oxidative stress in injured spinal cords. The mechanism may at least in part involve the eNOS/Nrf2/HO‑1 pathway.

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1	Mitchell R, Harvey L, Stanford R and Close J: Health outcomes and costs of acute traumatic spinal injury in New South Wales, Australia. Spine J. 2017.(Epub Ahead of Print). PubMed/NCBI
2	Moshi H, Sundelin G, Sahlen KG and Sorlin A: Traumatic spinal cord injury in the north-east Tanzania-describing incidence, etiology and clinical outcomes retrospectively. Global health Action. 10:13556042017. View Article : Google Scholar : PubMed/NCBI
3	Bothig R, Fiebag K, Thietje R, Faschingbauer M and Hirschfeld S: Morbidity of urinary tract infection after urodynamic examination of hospitalized SCI patients: The impact of bladder management. Spinal Cord. 51:70–73. 2013. View Article : Google Scholar : PubMed/NCBI
4	Pannek J: Editorial Note on: Morbidity of urinary tract infection after urodynamic examination of hospitalized SCI patients: The impact of bladder management. Spinal Cord. Sep 18–2012.(Epub Ahead of Print).
5	Borges TJ, Lang BJ, Lopes RL and Bonorino C: Modulation of alloimmunity by heat shock proteins. Front Immunol. 7:3032016. View Article : Google Scholar : PubMed/NCBI
6	Ren J, Fan C, Chen N, Huang J and Yang Q: Resveratrol pretreatment attenuates cerebral ischemic injury by upregulating expression of transcription factor Nrf2 and HO-1 in rats. Neurochem Res. 36:2352–2362. 2011. View Article : Google Scholar : PubMed/NCBI
7	Chini MG, Malafronte N, Vaccaro MC, Gualtieri MJ, Vassallo A, Vasaturo M, Castellano S, Milite C, Leone A, Bifulco G, et al: Identification of limonol derivatives as heat shock protein 90 (Hsp90) Inhibitors through a multidisciplinary approach. Chemistry. 22:13236–13250. 2016. View Article : Google Scholar : PubMed/NCBI
8	Dong X, Zheng L, Lu S and Yang Y: Neuroprotective effects of pretreatment of ginsenoside Rb1 on severe cerebral ischemia-induced injuries in aged mice: Involvement of anti-oxidant signaling. Geriatr Gerontol Int. 17:338–345. 2017. View Article : Google Scholar : PubMed/NCBI
9	Chen W, Guo Y, Yang W, Zheng P, Zeng J and Tong W: Involvement of connexin40 in the protective effects of ginsenoside Rb1 against traumatic brain injury. Cell Mol Neurobiol. 36:1057–1065. 2016. View Article : Google Scholar : PubMed/NCBI
10	Cheng W, Wu D, Zuo Q, Wang Z and Fan W: Ginsenoside Rb1 prevents interleukin-1 beta induced inflammation and apoptosis in human articular chondrocytes. Int Orthop. 37:2065–2070. 2013. View Article : Google Scholar : PubMed/NCBI
11	Liu DH, Chen YM, Liu Y, Hao BS, Zhou B, Wu L, Wang M, Chen L, Wu WK and Qian XX: Rb1 protects endothelial cells from hydrogen peroxide-induced cell senescence by modulating redox status. Biol Pharm Bull. 34:1072–1077. 2011. View Article : Google Scholar : PubMed/NCBI
12	Gurcan O, Gurcay AG, Kazanci A, Senturk S, Bodur E, Karaca EU, Turkoglu OF and Bavbek M: Effect of asiatic acid on the treatment of spinal cord injury: An Experimental study in rats. Turk Neurosurg. 27:259–264. 2017.PubMed/NCBI
13	Mo YP, Yao HJ, Lv W, Song LY, Song HT, Yuan XC, Mao YQ, Jing QK, Shi SH and Li ZG: Effects of electroacupuncture at governor vessel acupoints on neurotrophin-3 in rats with experimental spinal cord injury. Neural Plast. 2016:23718752016. View Article : Google Scholar : PubMed/NCBI
14	Armstrong HK, Koay YC, Irani S, Das R and Nassar ZD: Australian Prostate Cancer BioResource, Selth LA, Centenera MM, McAlpine SR and Butler LM: A novel class of Hsp90 C-terminal modulators have pre-clinical efficacy in prostate tumor cells without induction of a heat shock response. Prostate. 76:1546–1559. 2016. View Article : Google Scholar : PubMed/NCBI
15	Allen AR: Surgery of experimental lesion of spinal cord equivalent to crush injury of fracture dislocation of spinal column. J American Med Asso lvii. 878–880. 1962.
16	Allen AR and Reginald A: Remarks on the histopathological changes in the spinal cord due to impact. An experimental study. J Nervous Mental Dis. 41:141–147. 1914. View Article : Google Scholar
17	Basso DM, Beattie MS and Bresnahan JC: A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma. 12:1–21. 1995. View Article : Google Scholar : PubMed/NCBI
18	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 : PubMed/NCBI
19	Oh JH, Hyun JY and Varshavsky A: Control of Hsp90 chaperone and its clients by N-terminal acetylation and the N-end rule pathway. Proc Natl Acad Sci USA. 114:E4370–E4379. 2017. View Article : Google Scholar : PubMed/NCBI
20	Krzemien-Ojak L, Goral A, Joachimiak E, Filipek A and Fabczak H: Interaction of a novel chaperone PhLP2A with the heat shock protein Hsp90. J Cell Biochem. 118:420–429. 2017. View Article : Google Scholar : PubMed/NCBI
21	Bryan HK, Olayanju A, Goldring CE and Park BK: The Nrf2 cell defence pathway: Keap1-dependent and -independent mechanisms of regulation. Biochem Pharmacol. 85:705–717. 2013. View Article : Google Scholar : PubMed/NCBI
22	Kang CH, Kim MJ, Seo MJ, Choi YH, Jo WS, Lee KT, Jeong YK and Kim GY: 5-Hydroxy-3,6,7,8,3′4′-hexamethoxyflavone inhibits nitric oxide production in lipopolysaccharide-stimulated BV2 microglia via NF-κB suppression and Nrf-2-dependent heme oxygenase-1 induction. Food Chem Toxicol. 57:119–125. 2013. View Article : Google Scholar : PubMed/NCBI
23	Terazawa R, Akimoto N, Kato T, Itoh T, Fujita Y, Hamada N, Deguchi T, Iinuma M, Noda M, Nozawa Y and Ito M: A kavalactone derivative inhibits lipopolysaccharide-stimulated iNOS induction and NO production through activation of Nrf2 signaling in BV2 microglial cells. Pharmacol Res. 71:34–43. 2013. View Article : Google Scholar : PubMed/NCBI
24	Kim HJ, Zheng M, Kim SK, Cho JJ, Shin CH, Joe Y and Chung HT: CO/HO-1 induces NQO-1 expression via Nrf2 activation. Immune Netw. 11:376–382. 2011. View Article : Google Scholar : PubMed/NCBI
25	Li WC, Jiang DM, Hu N, Qi XT, Qiao B and Luo XJ: Lipopolysaccharide preconditioning attenuates neuroapoptosis and improves functional recovery through activation of Nrf2 in traumatic spinal cord injury rats. Int J Neurosci. 123:240–247. 2013. View Article : Google Scholar : PubMed/NCBI
26	Li J, Johnson D, Calkins M, Wright L, Svendsen C and Johnson J: Stabilization of Nrf2 by tBHQ confers protection against oxidative stress-induced cell death in human neural stem cells. Toxicol Sci. 83:313–328. 2005. View Article : Google Scholar : PubMed/NCBI
27	Rubiolo JA, Mithieux G and Vega FV: Resveratrol protects primary rat hepatocytes against oxidative stress damage: Activation of the Nrf2 transcription factor and augmented activities of antioxidant enzymes. Eur J Pharmacol. 591:66–72. 2008. View Article : Google Scholar : PubMed/NCBI
28	Zhang J, Zhang Y, Liang C, Wang N, Zheng H and Wang J: Choline supplementation alleviates fluoride-induced testicular toxicity by restoring the NGF and MEK expression in mice. Toxicol Appl Pharmacol. 310:205–214. 2016. View Article : Google Scholar : PubMed/NCBI
29	Miller DM, Singh IN, Wang JA and Hall ED: Administration of the Nrf2-ARE activators sulforaphane and carnosic acid attenuates 4-hydroxy-2-nonenal-induced mitochondrial dysfunction ex vivo. Free Radic Biol Med. 57:1–9. 2013. View Article : Google Scholar : PubMed/NCBI
30	Duan W, Zhang R, Guo Y, Jiang Y, Huang Y, Jiang H and Li C: Nrf2 activity is lost in the spinal cord and its astrocytes of aged mice. In Vitro Cell Dev Biol Anim. 45:388–397. 2009. View Article : Google Scholar : PubMed/NCBI
31	Ohashi R, Yan S, Mu H, Chai H, Yao Q, Lin PH and Chen C: Effects of homocysteine and ginsenoside Rb1 on endothelial proliferation and superoxide anion production. J Surg Res. 133:89–94. 2006. View Article : Google Scholar : PubMed/NCBI
32	Xie XS, Liu HC, Yang M, Zuo C, Deng Y and Fan JM: Ginsenoside Rb1, a panoxadiol saponin against oxidative damage and renal interstitial fibrosis in rats with unilateral ureteral obstruction. Chin J Integr Med. 15:133–140. 2009. View Article : Google Scholar : PubMed/NCBI
33	Sun Q, Meng QT, Jiang Y and Xia ZY: Ginsenoside Rb1 attenuates intestinal ischemia reperfusion induced renal injury by activating Nrf2/ARE pathway. Molecules. 17:7195–7205. 2012. View Article : Google Scholar : PubMed/NCBI
34	Hwang YP and Jeong HG: Ginsenoside Rb1 protects against 6-hydroxydopamine-induced oxidative stress by increasing heme oxygenase-1 expression through an estrogen receptor-related PI3K/Akt/Nrf2-dependent pathway in human dopaminergic cells. Toxicol Appl Pharmacol. 242:18–28. 2010. View Article : Google Scholar : PubMed/NCBI