Tanaka N, Espey L L, Okamura H
Department of Biology, Trinity University, San Antonio, Texas 78284.
Biol Reprod. 1989 Apr;40(4):762-8. doi: 10.1095/biolreprod40.4.762.
This study quantifies ovarian blood volume in Wistar rats by measuring the optical density (414 nm) of hemoglobin in ovarian extracts and comparing this measurement to the optical density of known amounts of whole blood. Immature rats were primed with pregnant mare's serum gonadotropin (PMSG), 10 IU s.c., at 23 days of age. On Day 25, the ovulatory process was initiated by human chorionic gonadotropin (hCG), 10 IU s.c., and ova began to appear in the oviducts 10 h later. At 2-h intervals, the ovaries were extirpated and homogenized in 1.0 ml of 0.05 M tris (hydroxymethyl)aminomethane buffer (pH 7.4) for 30 s. Homogenates were centrifuged for 20 min and the supernatant fluids were analyzed with a Gilford RESPONSE UV/VIS spectrophotometer. The hemoglobin in these ovarian extracts had the same peak absorbance of 414 nm characteristic of oxyhemoglobin in whole blood taken by cardiac puncture of the rats. There was a linear relationship between the absorbance and the volume of whole blood in the samples. The volume of blood per ovary from groups of 8 rats was 0.60 +/- 0.07 microL at 0 h after hCG. The volume increased to 1.37 +/- 0.26 microL at 4 h after hCG and reached a peak of 4.55 +/- 0.72 microL at 10 h. Indomethacin treatment (0.3-10.0 mg/rat, s.c.) partially inhibited this 7-fold increase in ovarian blood volume. In conclusion, the increase in ovarian blood volume during ovulation may reflect the vasodilation and hyperemia that are characteristic of inflamed tissues.
本研究通过测量卵巢提取物中血红蛋白的光密度（414nm），并将该测量值与已知量全血的光密度进行比较，对Wistar大鼠的卵巢血容量进行量化。未成熟大鼠在23日龄时皮下注射10IU孕马血清促性腺激素（PMSG）进行预处理。在第25天，皮下注射10IU人绒毛膜促性腺激素（hCG）启动排卵过程，10小时后卵子开始出现在输卵管中。每隔2小时，摘除卵巢并在1.0ml 0.05M三（羟甲基）氨基甲烷缓冲液（pH 7.4）中匀浆30秒。匀浆物离心20分钟，上清液用吉尔福德RESPONSE紫外/可见分光光度计分析。这些卵巢提取物中的血红蛋白与通过大鼠心脏穿刺采集的全血中氧合血红蛋白具有相同的414nm特征峰吸光度。样品中吸光度与全血体积之间存在线性关系。hCG注射后0小时，8只大鼠一组的每个卵巢血容量为0.60±0.07μL。hCG注射后4小时，血容量增加到1.37±0.26μL，并在10小时达到峰值4.55±0.72μL。吲哚美辛处理（0.3 - 10.0mg/大鼠，皮下注射）部分抑制了卵巢血容量7倍的增加。总之，排卵期间卵巢血容量的增加可能反映了炎症组织特有的血管舒张和充血。
