诱导型Hsp70在外源性甲醛对小鼠肝肾氧化损伤中的保护作用
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摘要
为探讨外源性甲醛是否会对小鼠肝脏和肾脏的热休克蛋白70(heat shock protein 70, Hsp70)产生影响,选雄性Balb/c小鼠为研究对象,采用动态吸入方式染毒2周,每周5 d,每天8 h,随机分为空白对照组、3 mg·m-3甲醛染毒组。染毒结束后,计算肝脏和肾脏的脏体比,检测各器官中活性氧(reactive oxygen species, ROS)、丙二醛(malondialdehyde, MDA)、还原型谷胱甘肽(glutathione, GSH)的含量,同时用实时荧光定量PCR(real-time PCR, RT-PCR)法检测Hsp70的基因表达,并通过H&E染色观察形态学变化。结果表明:与对照组相比,2种器官的脏体比均显著下降(P<0.05或P<0.01); ROS、MDA含量上升、GSH含量下降,出现显著性差异(P<0.05或P<0.01); Hsp70基因表达水平下降且差异显著(P<0.05或P<0.01); H&E切片显示,甲醛暴露使肝脏的中央静脉窦的破损程度增加、细胞核变大,肾脏中肾小中肾小球基底膜变窄、体积缩小甚至分解。上述结果表明,甲醛暴露会对小鼠的的肝脏和肾脏造成不同程度的氧化损伤,小鼠可通过上调Hsp70的基因表达以增强机体的抗氧化能力。
In order to explore the role of heat shock protein 70(Hsp70) in the toxic effect of exogenous formaldehyde(FA), male Balb/c mice were exposed to FA(3 mg·m-3) or clean room air by nose-only inhalation for 8 h·d-1, 5 days per week for two weeks. At the next day of the last-day exposure, the live and kidney tissues were collected for the terminal detection. Compared with the control group, the organ coefficients of liver or kidney tissues in the FA group significantly decreased(P<0.05 or P <0.01). The content of reactive oxygen species(ROS) andmalondialdehyde(MDA) both significantly increased(P<0.05 or P<0.01), while the level of glutathione(GSH) decreased significantly(P<0.05 or P<0.01). Meanwhile, the gene expression level of Hsp70 was detected by real-time PCR(RT-PCR), and reduced significantly(P <0.05 or P <0.01). Pathological data of the H & E staining slices showed that FA exposure aggravated the damage on the central venous sinus, and the nucleuses were enlarged in the liver. What's more, the glomerular basement membrane of kidney became narrower, and its volume was reduced or even decomposed. The results indicated that the oxidative stress induced by FA in kidney and liver tissues could be reversed by up-regulating Hsp70 gene expression.
In order to explore the role of heat shock protein 70(Hsp70) in the toxic effect of exogenous formaldehyde(FA), male Balb/c mice were exposed to FA(3 mg·m-3) or clean room air by nose-only inhalation for 8 h·d-1, 5 days per week for two weeks. At the next day of the last-day exposure, the live and kidney tissues were collected for the terminal detection. Compared with the control group, the organ coefficients of liver or kidney tissues in the FA group significantly decreased(P<0.05 or P <0.01). The content of reactive oxygen species(ROS) andmalondialdehyde(MDA) both significantly increased(P<0.05 or P<0.01), while the level of glutathione(GSH) decreased significantly(P<0.05 or P<0.01). Meanwhile, the gene expression level of Hsp70 was detected by real-time PCR(RT-PCR), and reduced significantly(P <0.05 or P <0.01). Pathological data of the H & E staining slices showed that FA exposure aggravated the damage on the central venous sinus, and the nucleuses were enlarged in the liver. What's more, the glomerular basement membrane of kidney became narrower, and its volume was reduced or even decomposed. The results indicated that the oxidative stress induced by FA in kidney and liver tissues could be reversed by up-regulating Hsp70 gene expression.
引文
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[2] Mohammadi S. Effect of selenium on neurotoxicity in adult male mice exposed to formaldehyde[J]. Electronic Physician, 2014, 6(4):939-943
[3] Wu D, Jiang Z, Gong B, et al. Vitamin E reversed apoptosis of cardiomyocytes induced by exposure to high dose formaldehyde during mice pregnancy[J]. International Heart Journal, 2017, 58(5):769-777
[4] Abd-Elhakim Y M, Mohamed A A, Mohamed W A. Hemato-immunologic impact of subchronic exposure to melamine and/or formaldehyde in mice[J]. Journal of Immunotoxicology, 2016, 13(5):713-722
[5] Wen H, Yuan L, Wei C, et al. Effects of combined exposure to formaldehyde and benzene on immune cells in the blood and spleen in Balb/c mice[J]. Environmental Toxicology&Pharmacology, 2016, 45:265-273
[6] Wei H, Tan K, Sun R, et al. Aberrant production of Th1/Th2/Th17-related cytokines in serum of C57BL/6 mice after short-term formaldehyde exposure[J]. International Journal of Environmental Research&Public Health,2014, 11(10):10036-10050
[7] Liu D, Zheng Y, Li B, et al. Adjuvant effects of gaseous formaldehyde on the hyper-responsiveness and inflammation in a mouse asthma model immunized by ovalbumin[J]. Journal of Immunotoxicology, 2011, 8(4):305-314
[8] Lima L F, Murta G L, Bandeira A C, et al. Short-term exposure to formaldehyde promotes oxidative damage and inflammation in the trachea and diaphragm muscle of adult rats[J]. Annals of Anatomy, 2015, 202:45-51
[9] Ciftci G, Aksoy A, Cenesiz S, et al. Therapeutic role of curcumin in oxidative DNA damage caused by formaldehyde[J]. Microscopy Research&Technique, 2015, 78(5):391-395
[10] Halasi M, Váraljai R, Benevolenskaya E, et al. A novel function of molecular chaperone HSP70[J]. The Journal of Biological Chemistry, 2016, 291:142-148
[11] Truman A W, Kristjansdottir K, Wolfgeher D, et al. The quantitative changes in the yeast Hsp70 and Hsp90 interactomes upon DNA damage[J]. Data in Brief, 2015, 2(C):12-15
[12] Hu Y, Zhou Y, Yang G, et al. Sulforaphane-N-acetyl-cysteine inhibited autophagy leading to apoptosis via Hsp70-mediated microtubule disruption[J]. Cancer Letters, 2018,431:85-95
[13] Wang X R, Bin Y, Min S, et al. Overexpression of a Brassica campestris HSP70 in tobacco confers enhanced tolerance to heat stress[J]. Protoplasma, 2016, 253(3):637-645
[14] Yi H, Huang G, Zhang K, et al. HSP70 protects rats and hippocampal neurons from central nervous system oxygen toxicity by suppression of NO production and NF-κB activation[J]. Experimental Biology&Medicine, 2018, 243(9):770-779
[15] Kowalczyk M, Owczarek A, Suchanek R, et al. Heat shock protein 70 gene polymorphisms are associated with paranoid schizophrenia in the Polish population[J]. Cell Stress&Chaperones, 2014, 19(2):205-215
[16] Padmini E, Geetha B V. Impact of season on liver mitochondrial oxidative stress and the expression of HSP70 in grey mullets from contaminated estuary[J]. Ecotoxicology, 2009, 18(3):304-311
[17] Tang X, Bai Y, Duong A, et al. Formaldehyde in China:Production, consumption, exposure levels, and health effects[J]. Environment International, 2009, 35(8):1210-1224
[18]王爱华,李继连,张斌,等.苏丹红Ⅰ对小鼠血液和脏器系数的影响[J].食品科学, 2010, 31(5):279-282Wang A H, Li J L, Zhang B. Effect of Sudan I on blood and organ coefficients of mice[J]. Food Science, 2010, 31(5):279-282(in Chinese)
[19]安松兰,张善玉,朴惠顺. 1年生黄芪中黄芪多糖对小鼠免疫器官指数的影响[J].延边大学医学学报, 2007,30(1):20-22An S, Zhang S, Piao H, et al. Effects of polysaccharides from the annual Astragalus on immuno-organs coefficients on mice[J]. Journal of Medical Science Yanbian University, 2007, 30(1):20-22(in Chinese)
[20] Tang B, Liu X, Ma Y, et al. Measurement of organ weight, organ coefficients and blood indices in HSF_(1)knockout mice[J]. Chinese Journal of Laboratory Animal Science, 2002, 12(3):153-156
[21] Storz G, Imlay J A. Oxidative stress[J]. Current Opinion in Microbiology, 1999, 2(2):188-194
[22] You B R, Park W H. Auranofin induces mesothelioma cell death through oxidative stress and GSH depletion[J].Oncology Reports, 2015, 35(1):546-551
[23] Valkoa M, Rhodesb C J, Moncol J, et al. Free radicals,metals and antioxidants in oxidative stress-induced cancer[J]. Chemico-Biological Interactions, 2006, 160(1):1-40
[24] Zhu Y, Li J, Wu Z. Acute exposure of ozone induced pulmonary injury and the protective role of vitamin E through the Nrf2 pathway in Balb/c mice[J]. Toxicology Research, 2015, 5(1):268-277
[25] Nardai G, Ti C S, Csermely P. Molecular chaperones,stress proteins and redox homeostasis[J]. Biofactors,2010, 17(1-4):249-257
[26] Guo S, Wharton W, Moseley P, et al. Heat shock protein70 regulates cellular redox status by modulating glutathione-related enzyme activities[J]. Cell Stress&Chaperones, 2007, 12(3):245-254
[27]李晓泓,宋晓琳,徐莉莉,等.即刻针刺和“逆针”三阴交穴对痛经大鼠子宫HSP70、MDA及SOD的影响[J].中华中医药杂志, 2010(11):1860-1863Li X, Song X, Xu L, et al. The change of HSP70, MDA and SOD in uterus of dysmenorrhea rats and the intervention of acupuncture and preventive acupuncture on Sanyinjiao point[J]. China Journal of Traditional Chinese Medicine&Pharmacy, 2010(11):1860-1863(in Chinese)
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