玉米胚芽提取物干预对雄性大鼠生殖系统衰老影响的研究
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摘要
目的研究采用氯化铝和D-半乳糖建立大鼠衰老模型,观察玉米胚芽提取物延缓雄性生殖系统衰老的作用,探讨其作用机制,为玉米胚芽提取物改善生殖系统衰老的研究提供理论依据。方法 SPF级雄性成年Wistar大鼠90只,体重180～220 g。D-半乳糖溶液(60 mg/kg·bw)腹腔注射,氯化铝溶液(200 mg/kg·bw)灌胃,连续8周,建立大鼠联合染毒衰老模型,设立空白对照组,玉米胚芽提取物(胚元胶囊)对照组,衰老模型组与低、中和高剂量的胚元胶囊干预组。采用苏木素-伊红(HE)染色观察睾丸形态学变化;免疫组化测定睾丸Bax和Bcl-2蛋白表达水平;生化法测定谷胱甘肽-S转移酶等氧化应激相关指标;ELISA方法测定睾酮等血清激素水平;实时定量PCR检测睾丸中Nrf2 mRNA表达量。结果与空白对照组相比,衰老模型组睾丸间质细胞,管腔内细胞少且分布紊乱;而低、中和高剂干预量组脱落溶解现象减少。衰老模型组Bax蛋白表达上调而Bcl-2蛋白表达下调,低、中和高剂量干预组Bax表达下调,Bcl-2表达上调。与空白对照组相比,衰老模型组的Nrf2 mRNA表达下调;低、中和高剂量干预组Nrf2基因表达上调。结论玉米胚芽提取物可改善复合衰老模型所致雄性大鼠睾丸细胞稀疏脱落的现象。复合衰老模型导致雄性大鼠睾丸组织细胞凋亡增加,玉米胚芽提取物干预降低细胞凋亡水平。玉米胚芽提取物干预影响生殖系统氧化应激水平。
Objective The aging model of rats induced by AlCl_3 and D-galactose was established in this study, to observe the anti-aging effect of extract from maize embryo, and clarify the potential mechanism of extract from maize embryo on reproductive system aging. Methods Ninety male adult Wistar rats(grade SPF, 180～220 g) were administrated by intraperitoneal injection with D-galactose(60 mg/kg·bw) and gavaged with AlCl_3(200 mg/kg·bw) for 8 weeks to establish a combined aging model. The blank control group, Peiyuan Capsule control group, aging model group, the low, middle and high dose Peiyuan Capsule intervention group were included in the study. The morphological changes of testis were observed by HE staining. The expression levels of Bax and Bcl-2 in testis were determined by ICH and some indexes of oxidative stress such as GSTs were determined by biochemical assays. Testosterone and other serum hormone indicators were determined by ELISA. The level of Nrf2 mRNA in testis was detected by real-time PCR. Result Compared with the blank control group, the luminal cells of the seminiferous tubules in the testicular tissue were thinning and more disorganized in the aging model group. For the low, middle and high dose groups, the defluxion and dissolution of Leydig cells decreased. In the aging model group, expression level of Bax protein increased and the expression level of Bcl-2 protein was inhibited, while in low, middle and high dose groups, the expression level of Bcl-2 increased, and the expression level of Bax decreased. Compared with the control group, Nrf2 mRNA level in the aging model group was reduced, while in the low, middle and high dose groups, Nrf2 mRNA level was up-regulated, which can increase the antioxidant ability, and promote the resistance of the reproductive system to aging. Conclusion The intervention of extract from maize embryo can improve the sparsity and abscission of the testis cells of the male aging model rats. The intervention of extract from maize embryo can reduce apoptosis. The intervention of extract from maize embryo can regulate the aging process in reproductive system by the alteration of oxidative stress.
Objective The aging model of rats induced by AlCl_3 and D-galactose was established in this study, to observe the anti-aging effect of extract from maize embryo, and clarify the potential mechanism of extract from maize embryo on reproductive system aging. Methods Ninety male adult Wistar rats(grade SPF, 180～220 g) were administrated by intraperitoneal injection with D-galactose(60 mg/kg·bw) and gavaged with AlCl_3(200 mg/kg·bw) for 8 weeks to establish a combined aging model. The blank control group, Peiyuan Capsule control group, aging model group, the low, middle and high dose Peiyuan Capsule intervention group were included in the study. The morphological changes of testis were observed by HE staining. The expression levels of Bax and Bcl-2 in testis were determined by ICH and some indexes of oxidative stress such as GSTs were determined by biochemical assays. Testosterone and other serum hormone indicators were determined by ELISA. The level of Nrf2 mRNA in testis was detected by real-time PCR. Result Compared with the blank control group, the luminal cells of the seminiferous tubules in the testicular tissue were thinning and more disorganized in the aging model group. For the low, middle and high dose groups, the defluxion and dissolution of Leydig cells decreased. In the aging model group, expression level of Bax protein increased and the expression level of Bcl-2 protein was inhibited, while in low, middle and high dose groups, the expression level of Bcl-2 increased, and the expression level of Bax decreased. Compared with the control group, Nrf2 mRNA level in the aging model group was reduced, while in the low, middle and high dose groups, Nrf2 mRNA level was up-regulated, which can increase the antioxidant ability, and promote the resistance of the reproductive system to aging. Conclusion The intervention of extract from maize embryo can improve the sparsity and abscission of the testis cells of the male aging model rats. The intervention of extract from maize embryo can reduce apoptosis. The intervention of extract from maize embryo can regulate the aging process in reproductive system by the alteration of oxidative stress.
引文
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[7] Yunhui Z, Renshan G, Hardy MP.Androgen-forming stem leydig cells: identification, function and therapeutic potential[J].Disease Markers,2008,24(4-5):277-286.
[8] Gaumer S,Guénal I,Brun S,et al.Bcl-2 and Bax mammalian regulators of apoptosis are functional in Drosophila[J]. Cell Death Differ,2000,7(9):804-814.
[9] 董雅洁,高维娟.Bcl-2、bax、caspase-3在细胞凋亡中的作用及其关系[J].中国老年学杂志,2012,32(21):4828-4830.
[10] Wulf D,Schipper HM.Oxidative stress and aberrant signaling in aging and cognitive decline[J]. Aging Cell,2007,6(3):361-370.
[11] Raghunath A,Sundarraj K,Nagarajan R,et al.Antioxidant response elements:Discovery, classes, regulation and potential applications[J]. Redox Biol,2018,17:297-314.
[2] 耿广琴,杨雅丽,李扬,等.党参水提物对D-半乳糖致衰老模型小鼠肝、肾线粒体呼吸链复合体Ⅰ、Ⅳ酶活性及线粒体结构的影响[J].中医研究,2015,28(1):67-70.
[3] 李江源,李小鹰,李明,等.血清游离睾酮水平和睾酮分泌指数随年龄老化而降低[J].中华男科学杂志, 2006, 12(6):555-558.
[4] 任婷婷,石亚丽,李书国.玉米胚食品加工技术的研究进展[J].粮食与饲料工业,2012,12(5):28-30.
[5] 董文彦,马熙媛,张东平, 等.玉米胚的营养成分及其控制体重、预防心肌病变作用研究[J].食品科学,2003,24(6):132-135.
[6] 董文彦,马熙媛,张东平,等.玉米胚延缓衰老作用的研究[J].食品科学,2002,23(7):95-97.
[7] Yunhui Z, Renshan G, Hardy MP.Androgen-forming stem leydig cells: identification, function and therapeutic potential[J].Disease Markers,2008,24(4-5):277-286.
[8] Gaumer S,Guénal I,Brun S,et al.Bcl-2 and Bax mammalian regulators of apoptosis are functional in Drosophila[J]. Cell Death Differ,2000,7(9):804-814.
[9] 董雅洁,高维娟.Bcl-2、bax、caspase-3在细胞凋亡中的作用及其关系[J].中国老年学杂志,2012,32(21):4828-4830.
[10] Wulf D,Schipper HM.Oxidative stress and aberrant signaling in aging and cognitive decline[J]. Aging Cell,2007,6(3):361-370.
[11] Raghunath A,Sundarraj K,Nagarajan R,et al.Antioxidant response elements:Discovery, classes, regulation and potential applications[J]. Redox Biol,2018,17:297-314.