何首乌饮对衰老大鼠生精细胞线粒体凋亡通路关键基因表达的影响

安福丽 王玉娟 刘昊坤 杜云帆 秦瑄希 牛嗣云* 高福禄*

doi:10.16098/j.issn.0529-1356.2017.03.007

PDF(1534 KB)

解剖学报 ›› 2017, Vol. 48 ›› Issue (3) : 287-295. DOI: 10.16098/j.issn.0529-1356.2017.03.007

细胞和分子生物学

何首乌饮对衰老大鼠生精细胞线粒体凋亡通路关键基因表达的影响

安福丽¹王玉娟¹ 刘昊坤² 杜云帆²秦瑄希²牛嗣云^3* 高福禄^4*

作者信息 +

Effect of Heshouwuyin on regulating the key genes expression of mitochondrial apoptotic pathway in aging rats’spermatogenic cells#br#

AN Fu-li¹WANG Yu-juan¹ LIU Hao-kun²DU Yun-fan² QIN Xuan-xi² NIU Si-yun ^3* GAO Fu-lu^4*

Author information +

文章历史 +

摘要

目的探讨何首乌饮对衰老大鼠睾丸生精细胞线粒体凋亡通路关键基因表达的影响。方法选用12月龄Wistar雄鼠45只，随机分为3组：青年对照组、自然衰老组、何首乌饮组，每组15只，何首乌饮组和自然衰老组于16个月开始分别灌胃何首乌饮和等量蒸馏水，连续60d。青年对照组于12月龄，其余两组于18月龄分别进行实验，通过Real-time PCR、免疫荧光染色和免疫印迹法，观察各组大鼠睾丸组织线粒体凋亡通路关键基因DR6、BAX、Caspase-3、Cyt-C、14-3-3σ的表达变化。结果自然衰老大鼠线粒体凋亡通路关键基因14-3-3σ 表达明显低于青年对照组，DR6、BAX、Caspase-3、Cyt-C表达明显高于青年对照组；而何首乌饮用药后能明显逆转上述现象。结论何首乌饮提高衰老大鼠精子质量与线粒体凋亡通路有关。

Abstract

Objective To investigate the effect of Heshouwuyin on regulating the key genes expression of mitochondrial apoptotic pathway in aging rats’ spermatogenic cells. Methods Forty-five 12-month-old male Wistar rats were randomly divided into three groups: young control group(YCG), natural aging group(NAG) and Heshouwuyin group(SWYG), 15 rats per group. From 16monthold, SWYG and NAG were intragastrically given Heshouwuyin and equal amount of normal saline respectively. Gavage treatment kept for 60 days. YCG at 12-month-old and the other two groups at 18-month-old entered experiment respectively. Real-time PCR, immunofluorescence staining, and Western blotting were used to detect the expression of DR6, BAX, Caspase-3, Cyt-C and 14-3-3σ. Results The expression of 14-3-3σ, a key factor in the mitochondrial apoptotic pathway, was significantly lower and the expressions of DR6, BAX, Caspase-3, and Cytc were significantly higher in the NAG than in the YCG, but intervention with Heshouwuyin significantly reversed this phenomenon. Conclusion Heshouwuyin improves the spermatogenic function of the testes by regulating expression of key genes in the mitochondrial apoptosis pathway.

导出引用

安福丽王玉娟刘昊坤杜云帆秦瑄希牛嗣云高福禄. 何首乌饮对衰老大鼠生精细胞线粒体凋亡通路关键基因表达的影响[J]. 解剖学报. 2017, 48(3): 287-295 https://doi.org/10.16098/j.issn.0529-1356.2017.03.007

AN Fu-li WANG Yu-juan LIU Hao-kun DU Yun-fan QIN Xuan-xi NIU Si-yun GAO Fu-lu. Effect of Heshouwuyin on regulating the key genes expression of mitochondrial apoptotic pathway in aging rats’spermatogenic cells#br#[J]. Acta Anatomica Sinica. 2017, 48(3): 287-295 https://doi.org/10.16098/j.issn.0529-1356.2017.03.007

参考文献

［1］Luo RZh, Jia W, Zhao LB, et al.General review on Radix Plygoni Multif lori ［J］.Chinese Herbal Medicines, 2005,36（7）:142-145. (in Chinese)

罗瑞芝，贾伟，赵利斌，等.何首乌研究进展［J］. 中草药，2005，36（7）：142-145.

［2］Niu S, Chen J, Duan F, et al. Possible mechanism underlying the effect of Heshouwuyin, a tonifying kidney herb, on sperm quality in aging rats［J］.BMC Complementary and Alternative Medi, 2014, 14:250.

［3］Li MY, Han X. Mechanism of kidney asthenia and aging research ［J］. Journal of Liaoning University of Traditinal Chinese, 2011, 13(11):108-110. (in Chinese)

刘梦云，韩旭. 肾虚与衰老机理中西医研究概况［J］.辽宁中医药大学学报，2011，13（11）：108-110.

［4］Yuan D, Wang H, He H, et al. Protective effects of total flavonoids from Epimedium on the male mouse reproductive system against cyclophosphamide-induced oxidative injury by up-regulating the expressions of SOD3 and GPX1［J］.Phytother Res,2014,28(1):88-97.

［5］Yan X, Jiang Z, Bi L, et al. Salvianolic acid A attenuates TNF-α and D-GalN-induced ER stress-mediated and mitochondrial-dependent apoptosis by modulating Bax/Bcl-2 ratio and calcium release in hepatocyte LO2 cells［J］.Naunyn-Schmiedeberg’s Arch Pharmacol,2015,388(8):817-830.

［6］Guo Y, Niu SY, Gao FL, et al. Effect of Heshowuyin on Bax and Bcl-2 expression of testicular Leydig cells in the overtraining rat testis［J］. Lishizhen Medicine and Materia Medica Research, 2012,23(3):589-590. (in Chinese)

郭银，牛嗣云，高福禄，等.何首乌饮对过度训练大鼠睾丸间质细胞Bcl-2、Bax 的影响［J］.时珍国医国药，2012，23（3）：589-590.

［7］Chen J, Wang Y, Hui C,et al. Mechanisms of Heshouwuyin in regulating apoptosis of testicular cells in aging rats through mitochondrial pathway［J］.BMC Complement Altern Med,2016,16:337.

［8］Bott RC, Clopton DT, Fuller AM, et al. KDR-LacZ-expressing cells are involved in ovarian and testis-specific vascular development, suggesting a role for VEGFA in the regulation of this vasculature［J］.Cell Tissue Res,2010,342(1):117-130.

［9］Liu YL, Yang SS, Chen SJ, et al.OSR1 and SPAK cooperatively modulate Sertoli cell support of mouse spermatogenesis［J］.Sci Rep,2016,6:37205.

［10］Rao M, Xia W, Yang J, et al. Transient scrotal hyperthermia affects human sperm DNA integrity, sperm apoptosis, and sperm protein expression［J］.Andrology, 2016,4(6):1054-1063.

［11］Dai JJ, Niu YF, Wu CF, et al. Both death receptor and mitochondria mediated apoptotic pathways participated the occurrence of apoptosis in porcine vitrified mii stage oocytes［J］.Cryo Letters,2016,37(2):129-136.

［12］Xu W, Guo G, Li J, et al. Activation of Bcl-2-Caspase-9 apoptosis pathway in the testis of asthmatic mice［J］.PLoS One,2016,11(3):e0149353.

［13］Zhang ChCh, Zhou AF, Zhang ML, et al. Effects of experimental varicocele on the apoptosis of spermatogenic cells in rats ［J］. Zhonghua Nankeuxe, 2003, 9(7):507-511. (in Chinese)

张长城，周安方，张茂林，等．实验性精索静脉曲张对大鼠睾丸生精细胞凋亡的影响［J］．中国男科学杂志，2003，9（7）：507-511.

［14］Pan G, Bauer J H, Haridas V, et al. Identification and functional characterization of DR6,a novel death domain -containing TNF-receptor［J］.FEBS Lett,1998,431(3):351-356.

［15］Zeng L, Li T, Xu DC, et al. Death receptor 6 induces apoptosis not through type I or type II pathways, but via a unique mitochondria-dependent pathway by interacting with bax protein［J］.J Biol Chem,2012,287(34): 29125-29133.

［16］Jia Y, Lee KW, Swerdloff R,et al.Interaction of insulin-like growth factor-binding protein-3 and BAX in mitochondria promotes male germ cell apoptosis［J］. J Biol Chem,2010, 285(3):1726-1732.

［17］Harris MH, Thompson CB. The role of the Bcl-2 family in the regulation of outer mitochondrial membrane permeability［J］. Cell Death Differ,2000,7(12):1182-1191.

［18］Ricci JE, Munoz-Pinedo C, Fitzgerald P, et al. Disruption of mitochondrial function during apoptosis is mediated by Caspase cleavage of the p75 subunit of complex I of the electron transport chain［J］. Cell 2004,117(6):773-786.

［19］Lin L, Boran Ni, Hongmei Lin, et al. Traditional usages, botany, phytochemistry, pharmacology and toxicology of polygonum multiflorum thunb: a review［J］. J Ethnopharmacol, 2015,159: 158-183.

［20］Kuang R, Sun Y, Yuan W, et al. Protective effects of echinacoside, one of the phenylethanoid glycosides, on H2O2-induced cytotoxicity in PC12 cells［J］.Planta Med, 2009,75(14):1499-1504.

［21］Zhang D,Wang Z, Sheng C, et al. Icariin prevents amyloid beta-induced apoptosis via the PI3K/Akt pathway in PC-12 cells［J］. Evid Based Complement Alternat Med, 2015,2015:235-265.

［22］Li J, He C,Tong W, et al. Tanshinone IIA blocks dexamethasone-induced apoptosis in osteoblasts through inhibiting Nox4-derived ROS production［J］.Int J Clin Exp Pathol, 2015, 8(10): 13695-13706.

［23］Li XM, Han F, Shi YX. Mitochondrial apoptotic pathway is involved in apoptosis in hippocampus of post-traumatic stess disorder rats［J］.Acta Anatomica Sinica,2010,41,(2):201-205. (in Chinese)

李晓明，韩芳，石玉秀. 线粒体凋亡路径参与创伤后应激障碍大鼠海马神经元凋亡的调控［J］.解剖学报, 2010，41，（2）：201-205.