五味子乙素调控NF-κB通路对顺铂致HK-2细胞炎症损伤的保护作用
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摘要
目的研究五味子乙素对顺铂致HK-2细胞炎症损伤的保护作用及相关机制。方法以不同浓度的五味子乙素及顺铂干预人肾小管上皮HK-2细胞,CCK-8法检测细胞活性,Hoechst 33258荧光染色法观察细胞形态变化,Western blot法检测NF-κB活化,酶联免疫吸附试验(ELISA)法检测细胞上清液中炎性细胞因子[肿瘤坏死因子-α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)]水平,实时荧光聚合酶链反应(RT-PCR)法检测细胞中TNF-α,IL-1β,IL-6的m RNA表达。结果与对照组相比,顺铂组细胞活性显著下降,细胞形态明显改变;五味子乙素呈浓度依赖性地减轻顺铂的细胞毒性作用。同时,五味子乙素能显著抑制顺铂所致NF-κB活化及炎性细胞因子(TNF-α,IL-1β,IL-6)在细胞上清液中及m RNA表达的升高。结论五味子乙素能通过调控NF-κB通路对抗顺铂所致肾小管上皮细胞的炎症损伤。
Objective To investigate the protective effect of schisandrin B against Cisplatin-induced inflammation damage in HK-2cells and its related mechanism. Methods After treated with various concentrations of schisandrin B and Cisplatin,viability of HK-2cells was determined by CCK-8 assay; cellular morphology was evaluated by Hoechst 33258 staining; activation of NF-κB was detected by Western blot; levels of TNF-α,IL-1β,IL-6 in supernatants were measured by ELISA,m RNA expressions of TNF-α,IL-1β,IL-6 were detected by RT-PCR. Results Compared with the control group,the cell activity of the cisplatin group decreased significantly,and the cell morphology changed significantly. Schisandrin B dose-dependently reduced the cytotoxicity induced by cisplatin,at the same time,schisandrin B significantly inhibited the activation of NF-κB induced by Cisplatin and reversed the increase of levels of TNF-α,IL-1β,IL-6 in supernatants and m RNA expressions. Conclusion Schisandrin B can against Cisplatin-induced inflammation damage in HK-2 cells through regulating NF-κB pathway.
Objective To investigate the protective effect of schisandrin B against Cisplatin-induced inflammation damage in HK-2cells and its related mechanism. Methods After treated with various concentrations of schisandrin B and Cisplatin,viability of HK-2cells was determined by CCK-8 assay; cellular morphology was evaluated by Hoechst 33258 staining; activation of NF-κB was detected by Western blot; levels of TNF-α,IL-1β,IL-6 in supernatants were measured by ELISA,m RNA expressions of TNF-α,IL-1β,IL-6 were detected by RT-PCR. Results Compared with the control group,the cell activity of the cisplatin group decreased significantly,and the cell morphology changed significantly. Schisandrin B dose-dependently reduced the cytotoxicity induced by cisplatin,at the same time,schisandrin B significantly inhibited the activation of NF-κB induced by Cisplatin and reversed the increase of levels of TNF-α,IL-1β,IL-6 in supernatants and m RNA expressions. Conclusion Schisandrin B can against Cisplatin-induced inflammation damage in HK-2 cells through regulating NF-κB pathway.
引文
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[2]Fuertes MA,Castilla J,Alonso C,et al.Novel concepts in the development of platinum antitumor drugs[J].Curr Med Chem,2002,2(4):539-551.
[3]Ren D,Villeneuve NF,Jiang T,et al.Brusatol enhances the efficacy of chemotherapy by inhibiting the Nrf2-mediated defense mechanism[J].PNAS,2011,108(4):1433-1438.
[4]Luke DR,Vadiei K,Lopez-Berestein G.Role of vascular congestion in cisplatin-induced acute renal failure in the rat[J].Nephrol Dial Transplant,1992,7(1):1-7.
[5]Lee CK,Park KK,Chung AS,et al.Ginsenoside Rg3 enhances the chemosensitivity of tumors to cisplatin by reducing the basal level of nuclear factor erythroid 2-related factor 2-mediated heme oxygenase-1/NAD(P)H quinone oxidoreductase-1 and prevents normal tissue damage by scavenging cisplatin-induced intracellular reactive oxygen species[J].Food Chem Toxicol,2012,50(7):2565-2574.
[6]李梅,金晶,李佳,等.五味子乙素激活Nrf2/ARE通路对顺铂致HK-2细胞氧化应激损伤的保护作用[J].药学学报,2012,47(11):1434-1439.
[7]Peres LA,da Cunha AD Jr.Acute Nephrotoxicity of cisplatin:molecular mechanisms[J].J Bras Nefrol,2013,35(4):332-340.
[8]Amirshahrokhi K,Khalili AR.Thalidomide ameliorates cisplatininduced nephrotoxicity by inhibiting renal inflammation in an experimental model[J].Inflammation,2015,38(2):476-484.
[9]Miller RP,Tadagavadi RK,Ramesh G,et al.Mechanisms of Cisplatin nephrotoxicity[J].Toxins(Basel),2010,2(11):2490-2518.
[10]Ramesh G,Reeves WB.TNF-alpha mediates chemokine and cytokine expression and renal injury in cisplatin nephrotoxicity[J].J Clin Invest,2002,110(6):835-842.
[11]Zhu SH,Wang Y,Chen MW,et al.Protective effect of schisandrin B against cyclosporine A-induced nephrotoxicity in vitro and in vivo[J].Am J Chin Med,2012,40(3):551-566.
[12]Wu YF,Cao MF,Gao YP,et al.Down-modulation of heat shock protein 70 and up-modulation of Caspase-3 during schisandrin B-induced apoptosis in human hepatoma SMMC-7721cells[J].World J Gastroenterol,2004,10(20):2944-2948.
[13]Xu Y,Liu Z,Sun J,et al.Schisandrin B prevents doxorubicininduced chronic cardiotoxicity and enhances its anticancer activity in vivo[J].PLo S One,2011,6(12):e28335.
[14]Chiu PY,Leung HY,Ko KM.Schisandrin B enhances renal mitochondrial antioxidant status,functional and structural integrity,and protects against gentamicin-induced nephrotoxicity in rats[J].Biol Pharm Bull,2008,31(4):602-605.
[15]Checker R,Patwardhan RS,Sharma D,et al.Schisandrin B exhibits anti-inflammatory activity through modulation of the redox-sensitive transcription factors Nrf2 and NF-κB[J].Free Radic Biol Med,2012,53(7):1421-1430.