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补肾益精汤对弱精症大鼠精子线粒体功能的影响及其相关机制研究
王世平,李倩云,崔言秀,吕涛,徐光玉,王策正
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济南市人民医院泌尿外科济南 271100;山东淄博中心医院泌尿外科山东淄博 263000
摘要:
目的:研究补肾益精汤对大鼠精子线粒体呼吸功能及能量代谢的影响,并从基因水平分析其可能的相关机制。方法:随机数字表法将100 只SD 雄性大鼠分为正常对照组(A 组)、雷公藤多苷组(B 组)、生理盐水组(C 组)、左卡尼汀组(D组)及补肾益精汤组(E 组),每组20 只。A 组大鼠每日灌胃生理盐水 [1 mL/(kg · d)],B、C、D、E 组每日灌胃雷公藤多苷 [20 mg/(kg · d)],共21 d,处死A、B 两组大鼠,然后C 组每日灌胃生理盐水[1 mL/(kg · d)],D 组每日灌胃左卡尼汀[50 mg/(kg · d)],E 组每日灌胃补肾益精汤[3 g/(kg · d)],共35 d,处死C、D、E 三组大鼠。获取各组大鼠精液标本,检测精液精子运动参数、细胞线粒体呼吸控制率(RCR)及精子细胞ATP 含量,同时检测各组精子细胞特异性钙通道CatSper1基因含量,分析E 组RCR、ATP 分别与CatSper1 RNA 含量的相关性。结果:与A 组比较,B 组的精子密度、a+b 级百分率及存活率均明显下降(P <0.01)。与C 组比较,D 组、E 组a+b 级精子百分率明显提高(P <0.01)。RCR、ATP 含量及CatSper1 RNA 均提高。D、E 组的精子RCR 分别为(4.39±0.42 vs 5.15±0.39)μmol · min-1 · g-1,ATP 含量分别为(4.02±0.50 vs 5.82±0.40)μg/g,CatSper1 RNA 分别为(2.18±0.28 vs 2.64±0.28)ng/μL,差异均具有统计学意义(P <0.01)。E 组大鼠的RCR、ATP 含量与CatSper1 RNA 均呈正相关(r =0.912、0.893,P < 0.05)。结论:雷公藤多苷可成功制备弱精子症大鼠模型;补肾益精汤可明显提高弱精症大鼠的精子活动力,增加其精子呼吸功能及能量代谢,其机制可能与增加特异性钙通道CatSper1 RNA 基因的表达有关。
关键词:  补肾益精汤  弱精症  呼吸控制率  ATP  CatSper1
DOI:10.3969/j.issn.1007-6948.2020.06.004
投稿时间:2019-01-28
基金项目:国家自然科学青年基金(81202016);山东省中医药科技发展计划项目(2017-451)
Related Mechanism
WANG Shi-ping,LI Qian-yun,CUI Yan-xiu
Abstract:
Objective To study the effects of Bushen Yijing Decoction (BYD) on respiratory function and energy metabolism of sperm mitochondria in rats, and analyze the possible mechanism on the gene level. Methods One hundred male rats were randomly divided into control group (group A), Tripterygium wilfordii group (group B), saline group (group C), levocarnitine group (group D) and BYD group (group E),20 in each group. The rats in group A were given normal saline [1 mL/(kg · d)], while the rats in other groups were given Tripterygium wilfordii polysaccharide [20 mg/(kg · d)], by gavage all for 21 d. Then the rats in groups A and B were sacrificed, and the rats in groups C, D and E were ig given normal saline [1 mL/(kg · d)],levocarnitine [50 mg/(kg · d)], and BYD [3 g/(kg · d)] respectively, once daily, for 35 d. Then the rats in groups C, D and E were sacrificed. The sperm motility parameters, mitochondrial respiratory control rate (RCR) and the ATP content of sperm cells were measured, and the gene content of specific calcium channel CatSper1 in sperm cells in each group was detected. Meanwhile, the correlation among the three indicators above was analyzed. Results The percentage of sperm a+b and sperm survival rate were significantly lower in group B than those in group A (P <0.01). But there was no significant difference in sperm density between two groups (P>0.05). Compared with group C, the percentage of sperm a+b in groups D and E increased signi?cantly (P<0.01), and the contents of RCR, ATP and CatSper1 RNA also increased, while those in group E increased more obviously. The results of groups D and E were as follows: RCR (4.39 ±0.42 vs 5.15± 0.39) μmol · min-1 ·g-1, ATP content (4.02±0.50 vs 5.82±0.40) μg/g and CatSper1 RNA (2.18±0.28 vs 2.64±0.28) ng/μL, and the differences were statistically significant between the two groups (P<0.01). Conclusion Tripterygium wilfordii can made asthenospermia rats model successfully. BYD can significantly increase the sperm motility of asthenospermia rats and increase their function of sperm respiration and energy metabolism through increasing the expression of CatSper1 RNA gene.
Key words:  Bushen Yijing Decoction  asthenospermia  respiratory control rate  ATP  CatSper1

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