实验性大鼠左侧精索静脉曲张对睾丸Leydig细胞的影响
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摘要
目的 建立精索静脉曲张(Varicocele，VC)大鼠模型，研究VC对血清睾酮和睾丸内睾酮的影响；研究VC对睾丸Leydig细胞凋亡的影响，以探讨VC是否引起Leydig细胞总量的变化；研究Leydig细胞内StAR mRNA表达的变化，以探讨单个Leydig细胞合成睾酮能力是否降低。以确定VC是否对睾丸Leydig细胞形态、功能产生影响。
方法 1．购买青春期SD大鼠，分为4周对照组、4周实验组；8周对照组、8周实验组，每组各10只。通过部分结扎左肾静脉造成左侧精索静脉曲张模型，曲张精索内静脉内径增粗约3倍者作为实验组。4周、8周时处死大鼠取出睾丸称重，4％多聚甲醛／0.1M固定液固定，做HE染色。Image-pro plus 4.5彩色图像分析系统下测量曲细精管直径。
2．4、8周时处死大鼠，取静脉血用放免法测定血清睾酮浓度；取部分睾丸组织称重，按1／3(W／V)比例用甲醇匀浆；匀浆转入15ml离心管中，室温过夜；1800×g离心30分钟，提取上清液；烘干；以400ul二乙醚提纯二次，提纯液进行放免法测定。
3．部分睾丸经3％戊二醛预固定，1％四氧化锇再固定，丙酮逐级脱水，Epon812包埋，半薄切片光学定位，超薄切片，醋酸铀及枸橼酸铅双重染色，日立H-600IV型投射电镜观察。
Abjective To study the effect of experimental varicocele on the plasma testostorone and on the testicular testostorone in adolescent rats; To study the effect of experimental varicocele on the Leydig cell apoptosis and investigate the total numbers of Leydig cells; We examined whether the mRNA for steroidogenic acute regulatory (StAR) protein, a crucial factor in the rate-limiting step of aldosterone biosynthesis, is expressed lowly to investigate the steroidogenic capacities of individual Leydig cell. To ensure the effect of VC on the morphology and function of Leydig cells in the testis. Methods 1. A varicocele model was created by partial ligation of the left renal vein in adolescent SD rats. The Sprague-Dawley rats which spermatic veins triples were called experimental groups. The experimental groups or control groups have 10 rats respectively. Four and eight weeks after creation of the varicocele, the histology of the testes was studied. The testes were weighed and fixed by 4% paraformaldehyde. The diameters were measured with Image-pro plus software.
2. Serum T concentrations were measured using a RIA. Intratesticular testosterone concentration was measured for RIA. In brief, testes were homogenized in 70% methanol(testis weight/methanol volume=1/3) using homogenizer. The homogenates were transferred to 15ml screw cap testubes.
方法 1．购买青春期SD大鼠，分为4周对照组、4周实验组；8周对照组、8周实验组，每组各10只。通过部分结扎左肾静脉造成左侧精索静脉曲张模型，曲张精索内静脉内径增粗约3倍者作为实验组。4周、8周时处死大鼠取出睾丸称重，4％多聚甲醛／0.1M固定液固定，做HE染色。Image-pro plus 4.5彩色图像分析系统下测量曲细精管直径。
2．4、8周时处死大鼠，取静脉血用放免法测定血清睾酮浓度；取部分睾丸组织称重，按1／3(W／V)比例用甲醇匀浆；匀浆转入15ml离心管中，室温过夜；1800×g离心30分钟，提取上清液；烘干；以400ul二乙醚提纯二次，提纯液进行放免法测定。
3．部分睾丸经3％戊二醛预固定，1％四氧化锇再固定，丙酮逐级脱水，Epon812包埋，半薄切片光学定位，超薄切片，醋酸铀及枸橼酸铅双重染色，日立H-600IV型投射电镜观察。
Abjective To study the effect of experimental varicocele on the plasma testostorone and on the testicular testostorone in adolescent rats; To study the effect of experimental varicocele on the Leydig cell apoptosis and investigate the total numbers of Leydig cells; We examined whether the mRNA for steroidogenic acute regulatory (StAR) protein, a crucial factor in the rate-limiting step of aldosterone biosynthesis, is expressed lowly to investigate the steroidogenic capacities of individual Leydig cell. To ensure the effect of VC on the morphology and function of Leydig cells in the testis. Methods 1. A varicocele model was created by partial ligation of the left renal vein in adolescent SD rats. The Sprague-Dawley rats which spermatic veins triples were called experimental groups. The experimental groups or control groups have 10 rats respectively. Four and eight weeks after creation of the varicocele, the histology of the testes was studied. The testes were weighed and fixed by 4% paraformaldehyde. The diameters were measured with Image-pro plus software.
2. Serum T concentrations were measured using a RIA. Intratesticular testosterone concentration was measured for RIA. In brief, testes were homogenized in 70% methanol(testis weight/methanol volume=1/3) using homogenizer. The homogenates were transferred to 15ml screw cap testubes.
引文
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