Rhoifolin attenuates damage to hippocampal neuronal culture model of acquired epilepsy in vitro by regulating NF-κB/iNOS/COX-2 axis

Main Article Content

Huizhen Qi
Liang Liu


epilepsy, rhoifolin (ROF, apigenin 7-O-β-neohesperidoside), neuronal activity, apoptosis, NF-κB/iNOS/COX-2 axis


To assess the effect of Rhoifolin (ROF [apigenin 7-O-enneohesperidoside]) on the damage to hippocampal neuronal culture model of acquired epilepsy (AE) and investigate its possible mechanisms. A hippocampal neuronal culture model of AE was established through incubating HT-22 cells with MgCl2 free medium. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays were used to assess the effect of ROF on cell viability and apoptosis exposed to epilepsy. The oxidative stress and secretion of inflammatory cytokines were measured by reverse transcription-quantitative polymerase chain reaction and enzyme-linked-immunosorbent serologic assay, respectively. Immunoblot assays were performed to determine the protein expression levels of nuclear factor kappa B/nitric oxide synthases/cyclooxygenase-2 (NF-κB/iNOS/COX-2) axis. ROF increases viability and reduces apoptosis of AE medium-treated HT-22 cell line. ROF relieves oxidative stress in AE medium-treated HT-22 cell line. ROF decreases the levels of pro-inflammatory cytokines in AE medium-treated HT-22 cell line. The functional effects of ROF on AE medium- treated HT-22 cell line is through inhibiting NF-κB/iNOS/COX-2 axis. ROF increased viability, decreased apoptosis, suppressed oxidative stress, and reduced pro-inflammatory cytokine levels in an epilepsy model in vitro by inhibiting NF-κB/iNOS/COX-2 axis. ROF might serve as a potential drug for epilepsy treatment.

Abstract 406 | PDF Downloads 402 HTML Downloads 205 XML Downloads 18


Aoki, C., Takeuchi, Y., Higashi, K., Okamoto, Y., Nakanishi, A., Tandia, M., Uzawa, J., Ueda, K. and Moribe, K., 2017. Structural elucidation of a novel transglycosylated compound alpha-glucosyl rhoifolin and of alpha-glucosyl rutin by NMR spectroscopy. Carbohydrate Research 443–444: 37–41. 10.1016/j.carres.2017.03.011

Beltran-Corbellini, A., Aledo-Serrano, A., Moller, R.S., Perez-Palma, E., Garcia-Morales, I., Toledano, R. and Gil-Nagel, A., 2022. Epilepsy genetics and precision medicine in adults: a new landscape for developmental and epileptic encephalopathies. Frontiers in Neurology 13: 777115. 10.3389/fneur.2022.777115

Brinza, I., Abd-Alkhalek, A.M., El-Raey, M.A., Boiangiu, R.S., Eldahshan, O.A. and Hritcu, L., 2020. Ameliorative effects of rhoifolin in scopolamine-induced amnesic zebrafish (Danio rerio) model. Antioxidants (Basel) 9. 10.3390/antiox9070580

Chen, H., Qin, J., Shi, H., Li, Q., Zhou, S. and Chen, L., 2022. Rhoifolin ameliorates osteoarthritis via the Nrf2/NF-kappaB axis: in vitro and in vivo experiments. Osteoarthritis Cartilage. 10.1016/j.joca.2022.01.009

Coussio, J.D., 1964. Isolation of rhoifolin from Chorisia species (Bombacaceae). Experientia 20: 562. 10.1007/BF02150291

Cui, H. and Zhang, W., 2022. The neuroprotective effect of miR-136 on pilocarpine-induced temporal lobe epilepsy rats by inhibiting Wnt/beta-catenin signaling pathway. Computational and Mathematical Methods in Medicine 2022: 1938205. 10.1155/2022/1938205

d’Orio, P., Pelliccia, V., Biondi, D., Scarpa, P., Gozzo, F., Revay, M., Cardinale, F., Tassi, L. and Cossu, M., 2022. Surgery for tuberous sclerosis complex-related epilepsy: risk factors for an unfavorable seizure outcome. Seizure 97: 8–14. 10.1016/j.seizure.2022.02.013

Fang, J., Cao, Z., Song, X., Zhang, X., Mai, B., Wen, T., Lin, J., Chen, J., Chi, Y., Su, T. and Xiao, F., 2020. Rhoifolin alleviates inflammation of acute inflammation animal models and LPS-induced RAW264.7 cells via IKKbeta/NF-kappa-B signaling pathway. Inflammation 43: 2191–2201. 10.1007/s10753-020-01286-x

He, Y., Wu, Z., Qiu, C., Wang, X., Xiang, Y., Lu, T., He, Y., Shang, T., Zhu, Q., Wang, X., Zeng, Q., Zhang, H. and Li, D., 2019. Long non-coding RNA GAPLINC promotes angiogenesis by regulating miR-211 under hypoxia in human umbilical vein endothelial cells. Journal of Cellular and Molecular Medicine 23: 8090–8100. 10.1111/jcmm.14678

Koosha, S., Mohamed, Z., Sinniah, A. and Alshawsh, M.A., 2019. Investigation into the molecular mechanisms underlying the anti-proliferative and anti-tumorigenesis activities of diosmetin against HCT-116 human colorectal cancer. Scientific Reports 9: 5148. 10.1038/s41598-019-41685-1

Li, J., Qiu, C., Xu, P., Lu, Y. and Chen, R., 2020. Casticin improves respiratory dysfunction and attenuates oxidative stress and inflammation via inhibition of NF-kB in a chronic obstructive pulmonary disease model of chronic cigarette smoke-exposed rats. Drug Design Development and Therapy 14: 5019–5027. 10.2147/DDDT.S277126

Liang, T., Wu, J., Chen, H., Qian, J. and Xu, Z., 2022. Novel mutation of EPM2A causes progressive myoclonic epilepsy: a case report. Neurological Sciences 10.1007/s10072-022-05986-0

Liao, S., Song, F., Feng, W., Ding, X., Yao, J., Song, H., Liu, Y., Ma, S., Wang, Z., Lin, X., Xu, J., Zhao, J. and Liu, Q., 2019. Rhoifolin ameliorates titanium particle-stimulated osteolysis and attenuates osteoclastogenesis via RANKL-induced NF-kappaB and MAPK pathways. Journal of Cellular Physiology 234: 17600–17611. 10.1002/jcp.28384

Negm, W.A., El-Kadem, A.H., Elekhnawy, E., Attallah, N.G.M., Al-Hamoud, G.A., El-Masry, T.A. and Zayed, A., 2022. Wound-healing potential of rhoifolin-rich fraction isolated from Sanguisorba officinalis roots supported by enhancing re-epithelization, angiogenesis, anti-inflammatory, and anti-microbial effects. Pharmaceuticals (Basel) 15. 10.3390/ph15020178

Peng, S., Hu, C., Liu, X., Lei, L., He, G., Xiong, C. and Wu, W., 2020. Rhoifolin regulates oxidative stress and proinflammatory cytokine levels in Freund’s adjuvant-induced rheumatoid arthritis via inhibition of NF-kappaB. Brazilian Journal of Medical and Biological Research 53: e9489. 10.1590/1414-431x20209489

Torii, K., Ikegami, Y., Aoki, M., Kato, T., Hamakawa, T., Maruyama, T. and Yasui, T., 2022. Status epilepticus in a patient with intractable epilepsy caused by renal colic due to a ureter stone. IJU Case Reports 5: 85–87. 10.1002/iju5.12399

Wei, W., Yang, R., Zhang, J., Chen, H., Ye, J., Su, Q., Liao, J. and Xiao, Z., 2022. The mediating roles of family resilience and social support in the relationship between illness severity and depressive symptoms among primary caregivers of children with epilepsy in China. Frontiers in Neurology 13: 831899. 10.3389/fneur.2022.831899

Xiong, L., Lu, H., Hu, Y., Wang, W., Liu, R., Wan, X. and Fu, J., 2021. In vitro anti-motile effects of Rhoifolin, a flavonoid extracted from Callicarpa nudiflora on breast cancer cells via downregulating Podocalyxin-Ezrin interaction during epithelial mesenchymal transition. Phytomedicine 93: 153486. 10.1016/j.phymed.2021.153486

Xu, W., Zhang, W., Cui, L., Shi, L., Zhu, B., Lyu, T. J. and Ma, W., 2022. Novel mutation of SIK1 gene causing a mild form of pediatric epilepsy in a Chinese patient. Metabolic Brain Disease 10.1007/s11011-022-00943-4

Yan, J., Ni, B., Sheng, G., Zhang, Y., Xiao, Y., Ma, Y., Li, H., Wu, H. and Tu, C., 2021. Rhoifolin ameliorates osteoarthritis via regulating autophagy. Frontiers in Pharmacology 12: 661072. 10.3389/fphar.2021.661072

Yasue, M., Itaya, M., Inagaki, M., Katayama, H. and Kawamura, N., 1967. Studies on the constituents of Evodiopanax innovans nakai. I. Isolation of maltol and rhoifolin from the leaves. Yakugaku Zasshi 87: 247–250. 10.1248/yakushi1947.87.3_247