脱氧雪腐镰刀菌烯醇暴露对BeWo细胞炎症与凋亡相关基因表达的影响
|
摘要
目的观察脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)暴露对人胎盘绒毛癌细胞(BeWo细胞)炎症与凋亡相关基因表达的影响。方法将BeWo细胞进行培养和相应剂量(0～750 nmol/L)DON暴露,ELISA法检测人绒毛膜促性腺激素(β-HCG)水平,qPCR法检测基因白细胞介素(IL)-6、IL-8、肿瘤坏死因子(TNF)-α、环氧酶(Cox)-2、半胱氨酸天冬氨酸蛋白酶(Caspase)-3和B淋巴细胞瘤(Bcl)-xl的表达水平。结果与空白对照组相比,50 nmol/L DON暴露组炎症和凋亡相关基因的表达呈显著性改变,即炎症相关基因IL-6、TNF-α和Cox-2在3、12和24 h时表达均显著上调(P<0.01),而IL-8仅在暴露24 h时显著上调(P<0.01),而凋亡相关基因Caspase-3在3、12和24 h时表达均显著低于对照组(P<0.01),而Bcl-xl的表达在暴露24 h时显著高于对照组(P<0.01)。结论 BeWo细胞DON暴露时炎症反应增强,凋亡被抑制,推测以上结果可能是DON胚胎发育毒性的潜在分子机制之一。
OBJECTIVE To explore the effects of deoxynivalenol-induced mRNA expressions of inflammation and apoptosis in BeWo cells. METHODS After been cultured and transferred, BeWo cells were seeded for the CCK-8 assay. Then incubated with DON in the appropriate doses and time according to the CCK-8 result. ELISA kit was used to measure the β-HCG levels in BeWo cells. qPCR was used to measure mRNA expressions of inflammation(IL-6,IL-8,TNF-α and Cox-2) and apoptosis(Caspase-3 and Bcl-xl) in BeWo cells. RESULTS Compared with the control group, IL-6, TNF-α and Cox-2 significantly increased in BeWo cells which were exposed in 50 nmol/L DON in 3,12 and 24 h(P<0.01), while IL-8 only significantly increased in 24 h(P<0.01). Caspase-3 significantly decreased in 3,12 and 24 h(P<0.01),and Bcl-xl significantly increased in 24 h(P<0.01). CONCLUSION The change in the level of inflammation and apoptosis may be one of the critical molecular mechanisms for DON-induced embryotoxicity.
OBJECTIVE To explore the effects of deoxynivalenol-induced mRNA expressions of inflammation and apoptosis in BeWo cells. METHODS After been cultured and transferred, BeWo cells were seeded for the CCK-8 assay. Then incubated with DON in the appropriate doses and time according to the CCK-8 result. ELISA kit was used to measure the β-HCG levels in BeWo cells. qPCR was used to measure mRNA expressions of inflammation(IL-6,IL-8,TNF-α and Cox-2) and apoptosis(Caspase-3 and Bcl-xl) in BeWo cells. RESULTS Compared with the control group, IL-6, TNF-α and Cox-2 significantly increased in BeWo cells which were exposed in 50 nmol/L DON in 3,12 and 24 h(P<0.01), while IL-8 only significantly increased in 24 h(P<0.01). Caspase-3 significantly decreased in 3,12 and 24 h(P<0.01),and Bcl-xl significantly increased in 24 h(P<0.01). CONCLUSION The change in the level of inflammation and apoptosis may be one of the critical molecular mechanisms for DON-induced embryotoxicity.
引文
[1] KUCA K, POHANKA M. Chemical warfare agents[J]. Exs, 2010, 100: 543-558.
[2] YU M, CHEN L, PENG Z, et al. Embryotoxicity caused by DON-induced oxidative stress mediated by Nrf2/HO-1 pathway[J]. Toxins, 2017, 9(6):E188.
[3] HUSSAIN S P, HARRIS C C. Inflammation and cancer: an ancient link with novel potentials[J]. Int J Cancer, 2007, 121: 2373-2380.
[4] PARK S Y, JIN M L, YI E H, et al. Neochlorogenic acid inhibits against LPS-activated inflammatory responses through up-regulation of Nrf2/HO-1 and involving AMPK pathway[J]. Environ Toxicol Phar, 2018, 62: 1-10.
[5] RUIZ-MIYAZAWA K W, PINHO-RIBEIRO F A, BORGHI S M, et al. Hesperidin methylchalcone suppresses experimental gout arthritis in mice by inhibiting NF-kappaB activation[J]. J Agr Food Chem, 2018, 66: 6269-6280.
[6] WANG R, WANG J, SONG F, et al. Tanshinol ameliorates CCl4-induced liver fibrosis in rats through the regulation of Nrf2/HO-1 and NF-kappaB/IkappaBalpha signaling pathway[J]. Drug Des Dev Ther, 2018, 12:1281-1292.
[7] SALEEM M, QADIR M I, PERVEEN N, et al. Inhibitors of apoptotic proteins: new targets for anticancer therapy[J]. Chem Biol Drug Des, 2013, 82: 243-251.
[8] WU Q, DOHNAL V, KUCA K, et al. Trichothecenes: structure-toxic activity relationships[J]. Curr Drug Metab, 2013, 14: 641-660.
[9] WEIDNER M, WELSCH T, HUBNER F, et al. Identification and apoptotic potential of T-2 toxin metabolites in human cells[J]. J Agr Food Chem, 2012, 60: 5676-5684.
[10] SPANIC V, MARCEK T, ABICIC I, et al. Effects of fusarium head blight on wheat grain and malt infected by fusarium culmorum[J]. Toxins,2017, 10(1):E17.
[11] YU M, CHEN L, PENG Z, et al. Mechanism of deoxynivalenol effects on the reproductive system and fetus malformation: current status and future challenges[J]. Toxicol in vitro, 2017, 41:150-158.
[12] ADESSO S, AUTORE G, QUARONI A, et al. The Food contaminants nivalenol and deoxynivalenol induce inflammation in intestinal epithelial cells by regulating reactive oxygen species release[J]. Nutrients,2017, 9 (12):E1343.
[13] MOON Y, PESTKA J J. Cyclooxygenase-2 mediates interleukin-6 upregulation by vomitoxin (deoxynivalenol) in vitro and in vivo[J]. Toxicol Appl Pharm, 2003, 187: 80-88.
[14] KINSER S, JIA Q, LI M, et al. Gene expression profiling in spleens of deoxynivalenol-exposed mice: immediate early genes as primary targets[J]. J Toxicol Env Heal A, 2004, 67: 1423-1441.
[15] CHOI B, JEONG S, CHO J, et al. Effects of oral deoxynivalenol exposure on immune-related parameters in lymphoid organs and serum of mice vaccinated with porcine parvovirus vaccine[J]. Mycotoxin Res, 2013, 29: 185-192.
[16] LANDGREN C A, HENDRICH S, KOHUT M L. Low-level dietary deoxynivalenol and acute exercise stress result in immunotoxicity in BALB/c mice[J]. J Immunotoxicol, 2006, 3: 173-178.
[17] LI M, CUFF C F, PESTKA J. Modulation of murine host response to enteric reovirus infection by the trichothecene deoxynivalenol[J]. Toxicol Sci, 2005, 87: 134-145.
[18] LI M, HARKEMA J R, CUFF C F, et al. Deoxynivalenol exacerbates viral bronchopneumonia induced by respiratory reovirus infection[J]. Toxicol Sci, 2007, 95: 412-426.
[19] AMUZIE C J, HARKEMA J R, PESTKA J J. Tissue distribution and proinflammatory cytokine induction by the trichothecene deoxynivalenol in the mouse: comparison of nasal vs. oral exposure[J]. Toxicology, 2008, 248: 39-44.
[20] DAI Y, XIE H, XU Y. Evaluation of deoxynivalenol-induced toxic effects on mouse endometrial stromal cells: cell apoptosis and cell cycle[J]. Biochem Bioph Res Co, 2017, 483: 572-577.
[21] PENG Z, CHEN L, NUSSLER A K, et al. Current sights for mechanisms of deoxynivalenol-induced hepatotoxicity and prospective views for future scientific research: a mini review[J]. J Appl Toxicol, 2017, 37: 518-529.
[22] KONIGS M, SCHWERDT G, GEKLE M, et al. Effects of the mycotoxin deoxynivalenol on human primary hepatocytes[J]. Mol nutr Food Res, 2008, 52: 830-839.
[23] TAKAKURA N, NESSLANY F, FESSARD V, et al. Absence of in vitro genotoxicity potential of the mycotoxin deoxynivalenol in bacteria and in human TK6 and HepaRG cell lines[J]. Food Chem Toxicol, 2014, 66: 113-121.
[2] YU M, CHEN L, PENG Z, et al. Embryotoxicity caused by DON-induced oxidative stress mediated by Nrf2/HO-1 pathway[J]. Toxins, 2017, 9(6):E188.
[3] HUSSAIN S P, HARRIS C C. Inflammation and cancer: an ancient link with novel potentials[J]. Int J Cancer, 2007, 121: 2373-2380.
[4] PARK S Y, JIN M L, YI E H, et al. Neochlorogenic acid inhibits against LPS-activated inflammatory responses through up-regulation of Nrf2/HO-1 and involving AMPK pathway[J]. Environ Toxicol Phar, 2018, 62: 1-10.
[5] RUIZ-MIYAZAWA K W, PINHO-RIBEIRO F A, BORGHI S M, et al. Hesperidin methylchalcone suppresses experimental gout arthritis in mice by inhibiting NF-kappaB activation[J]. J Agr Food Chem, 2018, 66: 6269-6280.
[6] WANG R, WANG J, SONG F, et al. Tanshinol ameliorates CCl4-induced liver fibrosis in rats through the regulation of Nrf2/HO-1 and NF-kappaB/IkappaBalpha signaling pathway[J]. Drug Des Dev Ther, 2018, 12:1281-1292.
[7] SALEEM M, QADIR M I, PERVEEN N, et al. Inhibitors of apoptotic proteins: new targets for anticancer therapy[J]. Chem Biol Drug Des, 2013, 82: 243-251.
[8] WU Q, DOHNAL V, KUCA K, et al. Trichothecenes: structure-toxic activity relationships[J]. Curr Drug Metab, 2013, 14: 641-660.
[9] WEIDNER M, WELSCH T, HUBNER F, et al. Identification and apoptotic potential of T-2 toxin metabolites in human cells[J]. J Agr Food Chem, 2012, 60: 5676-5684.
[10] SPANIC V, MARCEK T, ABICIC I, et al. Effects of fusarium head blight on wheat grain and malt infected by fusarium culmorum[J]. Toxins,2017, 10(1):E17.
[11] YU M, CHEN L, PENG Z, et al. Mechanism of deoxynivalenol effects on the reproductive system and fetus malformation: current status and future challenges[J]. Toxicol in vitro, 2017, 41:150-158.
[12] ADESSO S, AUTORE G, QUARONI A, et al. The Food contaminants nivalenol and deoxynivalenol induce inflammation in intestinal epithelial cells by regulating reactive oxygen species release[J]. Nutrients,2017, 9 (12):E1343.
[13] MOON Y, PESTKA J J. Cyclooxygenase-2 mediates interleukin-6 upregulation by vomitoxin (deoxynivalenol) in vitro and in vivo[J]. Toxicol Appl Pharm, 2003, 187: 80-88.
[14] KINSER S, JIA Q, LI M, et al. Gene expression profiling in spleens of deoxynivalenol-exposed mice: immediate early genes as primary targets[J]. J Toxicol Env Heal A, 2004, 67: 1423-1441.
[15] CHOI B, JEONG S, CHO J, et al. Effects of oral deoxynivalenol exposure on immune-related parameters in lymphoid organs and serum of mice vaccinated with porcine parvovirus vaccine[J]. Mycotoxin Res, 2013, 29: 185-192.
[16] LANDGREN C A, HENDRICH S, KOHUT M L. Low-level dietary deoxynivalenol and acute exercise stress result in immunotoxicity in BALB/c mice[J]. J Immunotoxicol, 2006, 3: 173-178.
[17] LI M, CUFF C F, PESTKA J. Modulation of murine host response to enteric reovirus infection by the trichothecene deoxynivalenol[J]. Toxicol Sci, 2005, 87: 134-145.
[18] LI M, HARKEMA J R, CUFF C F, et al. Deoxynivalenol exacerbates viral bronchopneumonia induced by respiratory reovirus infection[J]. Toxicol Sci, 2007, 95: 412-426.
[19] AMUZIE C J, HARKEMA J R, PESTKA J J. Tissue distribution and proinflammatory cytokine induction by the trichothecene deoxynivalenol in the mouse: comparison of nasal vs. oral exposure[J]. Toxicology, 2008, 248: 39-44.
[20] DAI Y, XIE H, XU Y. Evaluation of deoxynivalenol-induced toxic effects on mouse endometrial stromal cells: cell apoptosis and cell cycle[J]. Biochem Bioph Res Co, 2017, 483: 572-577.
[21] PENG Z, CHEN L, NUSSLER A K, et al. Current sights for mechanisms of deoxynivalenol-induced hepatotoxicity and prospective views for future scientific research: a mini review[J]. J Appl Toxicol, 2017, 37: 518-529.
[22] KONIGS M, SCHWERDT G, GEKLE M, et al. Effects of the mycotoxin deoxynivalenol on human primary hepatocytes[J]. Mol nutr Food Res, 2008, 52: 830-839.
[23] TAKAKURA N, NESSLANY F, FESSARD V, et al. Absence of in vitro genotoxicity potential of the mycotoxin deoxynivalenol in bacteria and in human TK6 and HepaRG cell lines[J]. Food Chem Toxicol, 2014, 66: 113-121.