槲皮素通过影响Nrf2-Keap1信号通路减缓镉中毒大鼠的生殖机能损伤

doi:10.11843/j.issn.0366-6964.2018.05.019

摘要/Abstract

摘要：

本试验旨在探索联合使用槲皮素（QE）可以减轻镉（Cd）致雄性大鼠生殖系统损伤及在此过程Nrf2-Keap1信号通路发挥的作用。试验选取32只3月龄、体重相近的SPF级Wistar雄性大鼠，随机分为4组，每组8只，分别为对照组[0.5 mL·（kg·d）^-1生理盐水]、Cd组[5 mg·（kg·d）^-1 CdCl₂]、QE组[15 mg·（kg·d）^-1QE]、Cd+QE组[5 mg·（kg·d）^-1 CdCl₂+15 mg·（kg·d）^-1 QE]，连续灌胃1个月。试验结束后，计算各组大鼠睾丸及附睾脏器系数，检测精子质量及睾丸组织抗氧化水平等变化，利用RT-qPCR及Western blot法检测Nrf2及其相关基因mRNA和蛋白的表达。结果表明：与对照组相比，Cd组大鼠生殖机能受损严重。与Cd组相比，Cd+QE组大鼠睾丸附睾脏器系数（P<0.01）和睾酮分泌量（P<0.05）增加明显；精子质量提高；睾丸组织中MDA、H₂O₂含量极显著降低（P<0.01），GSH含量（P<0.05）和CAT、T-SOD、GSH-Px酶活性（P<0.01）明显提高；Nrf2及其相关基因NQO1、γ-GCS、HO-1、GSH-Px的mRNA和蛋白的表达呈现明显上升趋势（P<0.01或P<0.05），而Keap1基因和蛋白质的表达显著降低（P<0.05）。综上，QE可能通过激活Nrf2-Keap1信号通路有效减轻Cd引起的大鼠生殖系统损伤。

Abstract:

The aim of this study was to investigate the effect of Quercetin (QE) on the reproductive system damage induced by cadmium (Cd) in male rats and the role of Nrf2-Keap1 signaling pathway in this process. A total 32 3-month-old SPF Wistar male rats were randomly divided into four groups with eight rats in each group:control group[0.5 mL·(kg·d)^-1 physiological saline], Cd group[5 mg·(kg·d)^-1 CdCl₂], QE group[15 mg·(kg·d)^-1 QE], Cd+ QE group[5 mg·(kg·d)^-1 CdCl₂+15 mg·(kg·d)^-1 QE], the study lasted for 30 days. At the end of the trial, the testis and epididymis organs coefficients were calculated, then the sperm quality and the levels of anti-oxidation of testis were measured. RT-qPCR and Western blot were used to detect the changes of mRNA and protein expressions of Nrf2 and its related genes, respectively. The results showed that the reproductive function of rats in Cd group was seriously damaged compared with the control group. Compared with Cd group, the coefficients of testis and epididymis (P<0.01) and the secretion testosterone of rats in Cd+QE group were significantly increased (P<0.05). The semen quality was improved, furthermore, the levels of MDA, H₂O₂ were significantly decreased (P<0.01) and the levels of GSH (P<0.05), CAT, T-SOD, GSH-Px (P<0.01) were significantly increased in Cd+QE group. The analysis showed that the mRNA and protein expression levels of Nrf2, HO-1, NQO1, GSH-Px, γ-GCS in testis were significantly increased (P<0.05 or P <0.01) and Keap1 mRNA and protein expression levels were significantly decreased (P <0.05) in Cd+QE group. In conclusion, QE can effectively alleviate the reproductive system damage of male rats induced by Cd through activating Nrf2-Keap1 signaling pathway.

中图分类号:

S856.9

何刘, 李慧, 周厚继, 佟翠, 王佳美, 李美琦, 杨淑华, 何剑斌. 槲皮素通过影响Nrf2-Keap1信号通路减缓镉中毒大鼠的生殖机能损伤[J]. 畜牧兽医学报, 2018, 49(5): 1037-1046.

HE Liu, LI Hui, ZHOU Hou-ji, TONG Cui, WANG Jia-mei, LI Mei-qi, YANG Shu-hua, HE Jian-bin. Protective Effects of Quercetin on the Reproductive Injury Induced by Cadmium through the Nrf2-Keap1 Signaling Pathway in Rats[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 1037-1046.

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https://www.xmsyxb.com/CN/Y2018/V49/I5/1037

参考文献

[1] LIU Y, SU C, ZHANG H, et al. Interaction of soil heavy metal pollution with industrialisation and the landscape pattern in Taiyuan city, China[J]. PLoS One, 2014, 9(9):e105798.
[2] LIN Y P, TENG T P, CHANG T K. Multivariate analysis of soil heavy metal pollution and landscape pattern in Changhua county in Taiwan[J]. Landsc Urban Plan, 2002, 62(1):19-35.
[3] AMAMOU F, NEMMICHE S, MEZIANE R K, et al. Protective effect of olive oil and colocynth oil against cadmium-induced oxidative stress in the liver of Wistar rats[J]. Food Chem Toxicol, 2015, 78:177-184.
[4] WINIARSKA-MIECZAN A, KRUSINSKI R, KWIECIEN M. Tannic Acid influence on lead and cadmium accumulation in the hearts and lungs of rats[J]. Adv Clin Exp Med, 2013, 22(5):615-620.
[5] 袁燕, 孙娅, 江辰阳, 等. 镉对大鼠大脑皮质神经细胞Bcl-2, Bax和Caspase-3 mRNA转录的影响[J]. 中国兽医学报, 2012, 32(11):1725-1729.
YUAN Y, SUN Y, JIANG C Y, et al. Influence of cadmium on Bcl-2, Bax and Caspase-3 mRNA transcription in rat's primary cultured cerebral cortical neurons[J]. Chinese Journal of Veterinary Science, 2012, 32(11):1725-1729. (in Chinese)
[6] LI R Y, LUO X, LI L B, et al. The protective effects of melatonin against oxidative stress and inflammation induced by acute cadmium exposure in mice testis[J]. Biol Trace Elem Res, 2016, 170(1):152-164.
[7] ERBOGA M, KANTER M, AKTAS C, et al. Anti-apoptotic and anti-oxidant effects of caffeic acid phenethyl ester on cadmium-induced testicular toxicity in Rats[J]. Biol Trace Elem Res, 2016, 171(1):176-184.
[8] 刘艳丰, 唐淑珍, 张文举, 等. 沙棘叶黄酮对阿勒泰羊生长性能、屠宰性能和血清生化指标的影响[J]. 畜牧兽医学报, 2014, 45(12):1981-1987.
LIU Y F, TANG S Z, ZHANG W J, et al. Effects of flavonoids of sea buckthorn leaves on growth performance, slaughter performance and serum biochemical indexes of Altay sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2014, 45(12):1981-1987. (in Chinese)
[9] 郭长江, 徐静, 韦京豫, 等. 我国常见蔬菜类黄酮物质的含量[J]. 营养学报, 2009, 31(2):185-190.
GUO C J, XU J, WEI J Y, et al. Flavonoid contents of common vegetables in China[J]. Acta Nutrimenta Sinica, 2009, 31(2):185-190. (in Chinese)
[10] GAO W N, PU L L, WEI J Y, et al. Comparison of dietary intakes of myricetin, quercetin, kaempferol, apigenin and luteolin from vegetables and fruits in Chinese adults from four different regions[J]. Acta Nutrimenta Sinica, 2014, 36(6):559-564. 高蔚娜, 蒲玲玲, 韦京豫, 等. 中国四地区成人蔬菜和水果中杨梅黄酮、槲皮素、坎二菲醇、芹菜配基和玉米黄酮摄入量的比较[J]. 营养学报, 2014, 36(6):559-564. (in English)
[11] 王敏, 刘保林, 国旭丹. 槲皮素及其代谢物抑制氧化应激与炎症[J]. 食品科学, 2013, 34(15):256-260.
WANG M, LIU B L, GUO X D. Quercetin and one of its metabolites inhibit reactive oxygen-species and inflammation[J]. Food Science, 2013, 34(15):256-260. (in Chinese)
[12] 申永春, 杨婷, 文富强, 等. 槲皮素对香烟暴露大鼠肺组织氧化应激和Nrf2表达的影响[J]. 西部医学, 2011, 23(11):2071-2073.
SHEN Y C, YANG T, WEN F Q, et al. The effect of quercetin on oxidative stress and Nrt2 expression in cigarette smoke exposed rat lungs[J]. Medical Journal of West China, 2011, 23(11):2071-2073. (in Chinese)
[13] 魏彩冰, 周莲娣, 张育珍, 等. 槲皮素通过核因子E2相关因子/抗氧化应答元件(Nrf2/ARE)信号通路发挥对免疫性肝损伤的保护作用[J]. 细胞与分子免疫学杂志, 2017, 33(3):300-304.
WEI C B, ZHOU L D, ZHANG Y Z, et al. Protective effect of quercetin against immunological liver injury through activating Nrf2/ARE signaling pathway[J]. Chinese Journal of Cellular and Molecular Immunology, 2017, 33(3):300-304. (in Chinese)
[14] ALKHEDAIDE A, ALSHEHRI Z S, SABRY A, et al. Protective effect of grape seed extract against cadmium-induced testicular dysfunction[J]. Mol Med Rep, 2016, 13(4):3101-3109.
[15] UNSAL C, KANTER M, AKTAS C, et al. Role of quercetin in cadmium-induced oxidative stress, neuronal damage, and apoptosis in rats[J]. Toxicol Ind Health, 2015, 31(12):1106-1115.
[16] STOHS S J, BAGCHI D, HASSOUN E, et al. Oxidative mechanisms in the toxicity of chromium and cadmium ions[J]. J Environ Pathol Toxicol Oncol, 2001, 20(2):77-88.
[17] VACA C E, WILHELM J, HARMS-RINGDAHL M. Interaction of lipid peroxidation products with DNA. A review[J]. Mutat Res/Rev Genet Toxicol, 1988, 195(2):137-149.
[18] 卞建春, 路浩, 梅莉, 等. 铅镉联合染毒对大鼠大脑皮质神经细胞的氧化损伤及NAC的保护效应[J]. 中国兽医科学, 2008, 38(9):805-809.
BIAN J C, LU H, MEI L, et al. Lipid peroxidation and protective effect of NAC on lead and/or cadmium in cerebral cortical neurons of rats[J]. Chinese Veterinary Science, 2008, 38(9):805-809. (in Chinese)
[19] YANG S H, LONG M, YU L H, et al. Sulforaphane prevents testicular damage in kunming mice exposed to cadmium via activation of Nrf2/ARE signaling pathways[J]. Int J Mol Sci, 2016, 17(10):1703.
[20] 高蔚娜, 郭长江, 韦京豫, 等. 槲皮素对氧化应激大鼠肝细胞的保护作用[J]. 中国临床营养杂志, 2007, 15(4):209-213.
GAO W N, GUO C J, WEI J Y, et al. Protective effects of quercetin on primary cultured rat hepatocytes against oxidative stress[J]. Chinese Journal of Clinical Nutrition, 2007, 15(4):209-213. (in Chinese)
[21] 刘连, 徐园园, 许洁, 等. 氟对大鼠自由基、氧化应激及超微结构影响[J]. 中国公共卫生, 2009, 25(9):1114-1116.
LIU L, XU Y Y, XU J, et al. Effect of fluoride on free radical, oxidative stress and ultrastructure of liver in rats[J]. Chinese Journal of Public Health, 2009, 25(9):1114-1116. (in Chinese)
[22] 叶煦亭, 沙继宏, 周雍. 镉离子诱发的氧自由基对肝细胞损伤及钙离子分布的影响[J]. 中华预防医学杂志, 2000, 34(4):256.
YE X T, SHA J H, ZHOU Y. Effect of oxygen free radical induced by cadmium ion on hepatocyte injury and distribution of calcium[J]. Chinese Journal of Preventive Medicine, 2000, 34(4):256. (in Chinese)
[23] 张克烽, 张子平, 陈芸, 等. 动物抗氧化系统中主要抗氧化酶基因的研究进展[J]. 动物学杂志, 2007, 42(2):153-160.
ZHANG K F, ZHANG Z P, CHEN Y, et al. Antioxidant defense system in animals[J]. Chinese Journal of Zoology, 2007, 42(2):153-160. (in Chinese)
[24] 方春华, 乔琨, 刘智禹, 等. 海洋生物中抗氧化酶的研究进展[J]. 渔业研究, 2016, 38(4):331-342.
FANG C H, QIAO K, LIU Z Y, et al. The research progress of antioxidant enzymes in marine organisms[J]. Journal of Fisheries Research, 2016, 38(4):331-342. (in Chinese)
[25] NNA V U, UJAH G A, MOHAMED M, et al. Cadmium chloride-induced testicular toxicity in male wistar rats; prophylactic effect of quercetin, and assessment of testicular recovery following cadmium chloride withdrawal[J]. Biomed Pharmacother, 2017, 94:109-123.
[26] SYKIOTIS G P, HABEOS I G, SAMUELSON A V, et al. The role of the antioxidant and longevity-promoting Nrf2 pathway in metabolic regulation[J]. Curr Opin Clin Nutr Metab Care, 2011, 14(1):41-48.
[27] 李全, 张源, 梁跃伟, 等. Nrf2抗氧化应激保护内皮细胞损伤的研究进展[J]. 昆明理工大学学报:自然科学版, 2017, 42(2):77-81.
LI Q, ZHANG Y, LIANG Y W, et al. Research progress of Nrf2 regulate oxidative stress in vascular endothelial cells[J]. Journal of Kunming University of Science and Technology (Natural Science Edition), 2017, 42(2):77-81. (in Chinese)
[28] LIU Z P, XIANG Y Q, SUN G H. The KCTD family of proteins:structure, function, disease relevance[J]. Cell Biosci, 2013, 3:45.
[29] NAZIMABASHIR, MANOHARAN V, MILTONPRABU S. Cadmium induced cardiac oxidative stress in rats and its attenuation by GSP through the activation of Nrf2 signaling pathway[J]. Chem Biol Interact, 2015, 242:179-193.