高尿酸血症对小鼠生精功能和精子质量的影响及其机制

doi:10.11958/20221083

天津医药 ›› 2023, Vol. 51 ›› Issue (4): 376-381.doi: 10.11958/20221083

高尿酸血症对小鼠生精功能和精子质量的影响及其机制

江晓翠(), 田代志, 赵敏, 龚健, 余何, 姜兴宇, 萧闵()

湖北中医药大学中医药实验中心（邮编430065）

收稿日期:2022-07-08 修回日期:2022-11-10 出版日期:2023-04-15 发布日期:2023-04-20
通讯作者: 萧闵 E-mail:280185855@qq.com;531637551@qq.com
作者简介:江晓翠（1989），女，实验师，博士在读，主要从事动物模型及中西医结合防治不孕不育方面研究。E-mail：280185855@qq.com
基金资助:
国家自然科学基金青年项目(82104704);2021年度湖北省教育厅重点项目(D20212001)

Effects and mechanism of hyperuricemia on spermatogenesis and sperm quality in mice

JIANG Xiaocui(), TIAN Daizhi, ZHAO Min, GONG Jian, YU He, JIANG Xingyu, XIAO Min()

Experimental Center of Chinese Medicine, Hubei University of Chinese Medicine, Wuhan 430065, China

Received:2022-07-08 Revised:2022-11-10 Published:2023-04-15 Online:2023-04-20
Contact: XIAO Min E-mail:280185855@qq.com;531637551@qq.com

江晓翠, 田代志, 赵敏, 龚健, 余何, 姜兴宇, 萧闵. 高尿酸血症对小鼠生精功能和精子质量的影响及其机制[J]. 天津医药, 2023, 51(4): 376-381.

JIANG Xiaocui, TIAN Daizhi, ZHAO Min, GONG Jian, YU He, JIANG Xingyu, XIAO Min. Effects and mechanism of hyperuricemia on spermatogenesis and sperm quality in mice[J]. Tianjin Medical Journal, 2023, 51(4): 376-381.

摘要/Abstract

摘要：

目的探讨高尿酸血症（HUA）通过氧化损伤诱导睾丸细胞凋亡、降低小鼠生精功能和精子质量的机制。方法 36只雄性昆明小鼠随机分6组：氧嗪酸钾1 d、7 d、14 d组，对照1 d、7 d、14 d组。其中氧嗪酸钾组腹腔注射氧嗪酸钾悬液600 mg/（kg·d）。生化法检测血清尿酸（UA）、肌酐（Cre）、尿素氮（BUN）、丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、黄嘌呤氧化酶（XO）和睾丸组织超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、丙二醛（MDA）；HE染色法观察肝、肾、睾丸组织病理学变化并对睾丸HE切片进行生精功能评分；全自动精子分析仪检测精子密度和活动率；蛋白免疫印迹法检测睾丸组织B淋巴细胞瘤-2（Bcl-2）、Bcl-2相关蛋白X（Bax）、胱天蛋白酶3（Caspase-3）的表达。结果与各自对照组比较，氧嗪酸钾1 d组UA升高，氧嗪酸钾7 d组UA、BUN、AST、XO均升高，氧嗪酸钾14 d组UA、BUN、AST均升高（均P<0.01）。氧嗪酸钾7 d、14 d组肝、肾组织形态均有病变，氧嗪酸钾7 d组较14 d组轻微。与对照7 d组比较，氧嗪酸钾7 d组睾丸组织有明显病变，生精功能、精子密度和活动率降低（P<0.05），MDA含量升高，SOD、CAT活性降低（P<0.05），Bax、Caspase-3蛋白表达量及Bax/Bcl-2比例上升（P<0.05），Bcl-2蛋白表达量下降（P<0.05）。结论 HUA小鼠生精功能和精子质量降低，其机制可能为睾丸氧化损伤诱导细胞凋亡。

关键词: 高尿酸血症, 睾丸, 精液分析, 精子发生, 氧化性应激, 细胞凋亡

Abstract:

Objective To investigate the mechanism of hyperuricemia (HUA) inducing testicular cell apoptosis and reducing spermatogenesis and sperm quality in mice through oxidative damage. Methods Thirty-six Kunming male mice were divided into 6 groups by random number method: the potassium oxyazinate groups for 1 d, 7 d and 14 d, and the control groups for 1 d, 7 d and 14 d. The potassium oxyazinate group was intraperitoneally injected with potassium oxyazinate suspension 600 mg/(kg·d). Serum levels of uric acid (UA), creatinine (Cre), urea nitrogen (BUN), alanine aminotransferase (ALT), aspartate aminotransferase (AST), xanthine oxidase (XO), and superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) in testicular tissue were detected by biochemical method. Eosin-hematoxylin (HE) staining was used to observe histopathological changes of liver, kidney and testis, and the spermatogenic function of testicular HE sections was scored. Automatic sperm analyzer detected sperm density and motility rate. The expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and Caspase-3 protein in testicular tissue were detected by Western blotting. Results Compared with the control group, UA was significantly increased in the potassium oxazinate 1 d group (P<0.01). The levels of UA, BUN, AST and XO were significantly increased in the potassium oxazinate 7 d group (P<0.01). The levels of UA, BUN and AST were significantly increased in the potassium oxazinate 14 d group (P<0.01). The pathological changes in liver and kidney tissue were observed in the potassium oxazinate 7 d and 14 d groups, and the lesion was milder in the potassium oxazinate 7 d groug than that of the potassium oxazinate 14 d group. Compared with the control 7 d group, there were obvious pathological changes and significantly decreased spermatogenic function in the potassium oxazinate 7 d group (P<0.01). The content of MDA was significantly increased (P<0.05), while the activities of SOD and CAT were significantly decreased (P<0.05). The protein expression levels of Bax and Caspase-3 were significantly increased (P<0.05), the ratio of Bax to Bcl-2 was also increased (P<0.05), and the protein expression level of Bcl-2 was significantly decreased (P<0.05). Conclusion The spermatogenic function and sperm quality are decreased in HUA mice, which may be caused by testicular oxidative damage induced apoptosis.

Key words: hyperuricemia, testis, semen analysis, spermatogenesis, oxidative stress, apoptosis

中图分类号:

R698.2

图/表 9

表1 各组小鼠血清UA、BUN、Cre水平比较（n=6，$\bar{x} \pm s$）

Tab.1 The serum levels of UA, BUN and Cre in six groups of mice

组别	UA（μmol/L）	BUN（mmol/L）	Cre（μmol/L）
对照1 d组	38.65±9.36	3.30±0.75	43.95±3.73
氧嗪酸钾1 d组	147.86±37.33	4.00±0.73	47.45±4.63
t	7.009^**	1.634	1.444
对照7 d组	36.51±7.09	3.67±0.26	45.02±5.52
氧嗪酸钾7 d组	178.95±37.21	5.94±1.01	50.26±4.21
t	9.210^**	5.342^**	1.848
对照14 d组	39.15±7.94	4.46±0.65	45.40±2.37
氧嗪酸钾14 d组	190.97±53.07	6.90±1.03	48.75±3.63
t	6.930^**	6.947^**	1.887

表2 各组小鼠血清AST、ALT和XO水平比较（n=6，U/L，$\bar{x} \pm s$）

Tab.2 The serum levels of AST, ALT and XO in six groups of mice

组别	AST	ALT	XO
对照1 d组	109.33±18.25	20.83±2.54	30.76±2.61
氧嗪酸钾1 d组	123.78±16.26	21.01±1.87	32.71±2.08
t	1.448	0.140	1.432
对照7 d组	107.25±14.51	19.17±1.89	30.52±2.92
氧嗪酸钾7 d组	150.41±20.54	20.82±3.33	40.02±4.67
t	4.203^**	1.055	4.225^**
对照14 d组	107.59±16.76	20.47±2.05	32.50±1.63
氧嗪酸钾14 d组	165.93±19.57	22.98±3.24	37.26±5.00
t	5.546^**	1.602	2.215

图1 氧嗪酸钾不同注射天数对各组小鼠肝组织病理的影响（HE染色）

Fig.1 Effects of different days of potassium oxazinate injection on pathological changes of liver tissue in mice (HE staining)

图2 氧嗪酸钾不同注射天数对各组小鼠肾组织病理的影响（HE染色）

Fig.2 Effects of different days of potassium oxazinate injection on renal histopathological changes of mice in each group (HE staining)

图3 氧嗪酸钾7 d 对小鼠睾丸组织的影响（HE染色）

Fig.3 Effects of potassium oxazinate on testicular tissue of mice for 7 days (HE staining)

表3 2组小鼠睾丸生精功能变化（n=6，$\bar{x} \pm s$）

Tab.3 Changes of testicular spermatogenic function in the two groups

组别	D（μm）	M（μm）	P（分）	S（分）	TMS（分）
对照组	86.04±8.21	3.14±0.50	4.46±0.40	4.88±0.20	17.33±0.67
模型组	66.52±6.10	3.92±0.35	3.65±0.24	3.96±0.19	12.30±0.84
t	4.269^**	2.855^*	3.883^**	7.389^**	10.433^**

表4 2组小鼠精子密度和活动率变化（n=6，$\bar{x} \pm s$）

Tab.4 Changes of sperm density and motility in the two groups

组别	精子密度（×10⁶/mL）	精子活动率（%）
对照组	52.83±6.61	56.83±7.50
模型组	32.50±9.73	44.01±5.48
t	4.235^**	3.378^**

图4 2组小鼠睾丸组织氧化应激指标变化

Fig.4 Changes of oxidative stress index in testicular tissue in the two groups A：SOD；B：MDA；C：CAT；n=6，*P<0.05。

图5 2组小鼠睾丸组织Bax、Bcl-2、Caspase-3蛋白表达量变化 A：蛋白免疫印迹结果；B~E分别为组间Bax、Bcl-2、Caspase-3、Bax/Bcl-2比较；n=3，*P<0.05。

Fig.5 Changes of Bax, Bcl-2 and Caspase-3 protein expression in testicular tissue of the two groups

参考文献 22

[1]	LIU L, JIANG S, LIU X, et al. Inflammatory response and oxidative stress as mechanism of reducing hyperuricemia of gardenia jasminoide-poria cocos with network pharmacology[J]. Oxid Med Cell Longev, 2021, 2021:8031319. doi:10.1155/2021/8031319.
[2]	KIMURA Y, TSUKUI D, KONO H. Uric acid in inflammation and the pathogenesis of atherosclerosis[J]. Int J Mol Sci, 2021, 22(22):12394. doi:10.3390/ijms222212394.
[3]	PIAO W, ZHAO L, YANG Y, et al. The prevalence of hyperuricemia and its correlates among adults in China: Results from CNHS 2015-2017[J]. Nutrients, 2022, 14(19):4095. doi:10.3390/nu14194095.
[4]	左俊, 李坤, 邓大同, 等. 2017-2020年成人体检人群高尿酸血症患病率调查分析[J]. 安徽医专学报, 2021, 20(4):10-15.
	ZUO J, LI K, DENG D T, et al. Investigation and analysis of the prevalence of hyperuricemia in adult physical examination population from 2017 to 2020[J]. Journal of Anhui Medical College, 2021, 20(4):10-15. doi:10.3969/j.issn.2097-0196.2021.04.005.
[5]	LEISEGANG K, SENGUPTA P, AGARWAL A, et al. Obesity and male infertility:Mechanisms and management[J]. Andrologia, 2021, 53(1):e13617. doi:10.1111/and.13617.
[6]	FACONDO P, DI LODOVICO E, DELBARBA A, et al. The impact of diabetes mellitus type 1 on male fertility:Systematic review and meta-analysis[J]. Andrology, 2022, 10(3):426-440. doi:10.1111/andr.13140.
[7]	韩瑞钰, 马婧, 崔彤, 等. 高尿酸血症对不育男性精液参数的影响[J]. 中华男科学杂志, 2018, 24(12):1069-1072.
	HAN R Y, MA J, CUI T, et al. Hyperuricemia reduces semen parameters in infertile men[J]. Chinese Journal of Andrology, 2018, 24(12):1069-1072. doi:10.13263/j.cnki.nja.2018.12.003.
[8]	MA J, HAN R, CUI T, et al. Effects of high serum uric acid levels on oxidative stress levels and semen parameters in male infertile patients[J]. Medicine (Baltimore), 2022, 101(3):e28442. doi:10.1097/MD.0000000000028442.
[9]	LIN G, YU Q, XU L, et al. Berberrubine attenuates potassium oxonate- and hypoxanthine-induced hyperuricemia by regulating urate transporters and JAK2/STAT3 signaling pathway[J]. Eur J Pharmacol, 2021, 912:174592. doi: 10.1016/j.ejphar.2021.174592.
[10]	PAI H L, HSIEH S M, SU Y S, et al. Short-term hyperuricemia leads to structural retinal changes that can be reversed by serum uric acid lowering agents in mice[J]. Invest Ophthalmol Vis Sci, 2022, 63(10):8. doi:10.1167/iovs.63.10.8.
[11]	萧闵, 王威, 魏巍, 等. 疏肝补肾毓麟汤调节VDAC2基因甲基化影响弱精症大鼠精子线粒体功能改善精子质量[J]. 中国实验方剂学杂志, 2021, 27(23):72-79.
	XIAO M, WANG W, WEI W, et al. Shugan Bushen Yulin Decoction regulates VDAC2 gene methylation to affect sperm mitochondrial function and improve sperm quality in asthenospermia rats[J]. Chinese Journal of Experimental Traditional Medical Formulae, 2021, 27(23)72-79. doi:10.13422/j.cnki.syfjx.20212338.
[12]	YANG C, LI P, LI Z. Clinical application of aromatase inhibitors to treat male infertility[J]. Hum Reprod Update, 2021, 28(1):30-50. doi:10.1093/humupd/dmab036.
[13]	李昕, 李豫, 邵骏, 等. 中国有生育力男性精子浓度35年变化趋势分析[J]. 中华男科学杂志, 2021, 27(7):645-648.
	LI X, LI Y, SHAO J, et al. Trend of change in sperm concentration in Chinese fertile males from 1984 to 2019[J]. National Journal of Andrology, 2021, 27(7):645-648. doi:10.13263/j.cnki.nja.2021.07.012.
[14]	ANTINOZZI C, LISTA M, CAPONECCHIA L, et al. Exploratory analysis in the differences in blood serum and seminal plasma of adipose-tissue related peptides in obese and non-obese men and their correlations with semen parameters[J]. Front Endocrinol(Lausanne), 2021, 12:681939. doi:10.3389/fendo.2021.681939.
[15]	NETHENGWE M, OKAIYETO K, OGUNTIBEJU O O, et al. Ameliorative effects of Anchomanes difformis aqueous extract against oxidative stress in the testes and epididymis of streptozotocin-induced diabetic male Wistar rats[J]. Saudi J Biol Sci, 2022, 29(5):3122-3132. doi:10.1016/j.sjbs.2022.03.015.
[16]	NIU Y, TANG Q, ZHAO X, et al. Obesity-induced insulin resistance is mediated by high uric acid in obese children and adolescents[J]. Front Endocrinol (Lausanne), 2021, 12:773820. doi:10.3389/fendo.2021.773820.
[17]	CHEN M, JI H, SONG W, et al. Anserine beneficial effects in hyperuricemic rats by inhibiting XOD,regulating uric acid transporter and repairing hepatorenal injury[J]. Food Funct, 2022, 13(18):9434-9442. doi:10.1039/d2fo01533a.
[18]	WANG F, ZHAO X, SU X, et al. Isorhamnetin,the xanthine oxidase inhibitor from Sophora japonica,ameliorates uric acid levels and renal function in hyperuricemic mice[J]. Food Funct, 2021, 12(24):12503-12512. doi: 10.1039/d1fo02719k.
[19]	谭璐嫔, 唐凤, 韩萍萍, 等. 二甲双胍对高尿酸诱导HepG2细胞脂质沉积的作用及其机制研究[J]. 中华糖尿病杂志, 2021, 13(6):618-625.
	TAN L P, TANG F, HAN P P, et al. Effects of metformin on lipid deposition in HepG2 cells induced by high uric acid[J]. Chinese Journal of Diabetes Mellitus, 2021, 13(6):618-625. doi:10.3760/cma.j.cn115791-20210107-00009.
[20]	谢超, 张胜健, 谭嘉莉, 等. 血尿酸升高是导致肾功能下降的独立危险因素[J]. 中华肾脏病杂志, 2019, 35(2):100-105.
	XIE C, ZHANG S J, TAN J L, et al. Elevated serum uric acid is an independent risk factor for the loss of renal function[J]. Chinese Journal of Nephrology, 2019, 35(2):100-105. doi: 10.3760/cma.j.issn.1001-7097.2019.02.004.
[21]	乔美玲, 骆骄阳, 杨明翰, 等. 多伞阿魏对人胃癌MGC-803裸鼠移植瘤生长及凋亡的影响[J]. 中国中药杂志, 2019, 44(13):2827-2834.
	QIAO M L, LUO J Y, YANG M H, et al. Effect of Ferula ferulaeoides on growth and apoptosis of human gastric cancer MGC-803 transplantation tumor in nude mice[J]. China Journal of Chinese Materia Medica, 2019, 44(13):2827-2834. doi:10.19540/j.cnki.cjcmm.20190401.401.
[22]	KARAM Z M, BABA SALARI M, ANJOM SHOAA A, et al. Impact of oxidative stress SNPs on sperm DNA damage and male infertility in a south-east Iranian population[J]. Reprod Fertil Dev, 2022, 34(8):633-643. doi:10.1071/RD21305.

高尿酸血症对小鼠生精功能和精子质量的影响及其机制

Effects and mechanism of hyperuricemia on spermatogenesis and sperm quality in mice

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