Hixenbaugh E A, Sullivan T R, Strauss J F, Laposata E A, Komaromy M, Paavola L G
Department of Anatomy, Temple University School of Medicine, Philadelphia, Pennsylvania 19140.
J Biol Chem. 1989 Mar 5;264(7):4222-30.
Hepatic lipase is proposed to have a role in steroidogenesis through its involvement in the metabolism of high density lipoproteins. We examined the activity, synthesis, distribution, and uptake of this enzyme and assessed the content of its mRNA in luteinized ovaries. We found that during peak steroidogenesis, ovaries of pregnant mare's serum gonadotropin-human chorionic gonadotropin-treated immature rats contained heparin-releasable hepatic lipase-like activity which was neutralized in a dose-dependent manner by purified antibodies to hepatic lipase isolated from post-heparin perfusates of rat livers. Quantitative immunoelectron microscopy revealed that ovarian hepatic lipase occurred along endothelial cells and was 3-fold more abundant in blood vessels of corpora lutea than those of stroma. However, hepatic lipase was not synthesized by the ovary since radiolabeled enzyme was not immunoisolated from the medium of dispersed luteinized granulosa cells incubated with [35S]methionine whereas it was present in the medium of control cells (hepatocytes). Similarly, hepatic lipase mRNA was detectable in liver but not ovaries or kidneys by Northern or slot blot analyses or by the polymerase chain reaction. Finally, 125I-labeled hepatic lipase injected into tail veins was quickly cleared from the systemic circulation, accumulating in liver, ovaries, kidneys, and spleen. Subsequent heparin injection caused rapid reappearance of radioactivity in the bloodstream and a marked decline of radiolabel in liver and ovaries but a modest decrease of that in kidneys and none in spleen. Exogenous 125I-bovine serum albumin also accumulated in all four organs but was not displaced from liver or ovaries by subsequent administration of heparin. Taken together, these data suggest that steroidogenically active ovaries possess but do not synthesize hepatic lipase. Instead, hepatic lipase originating elsewhere, presumably in the liver, is accumulated from the circulation at heparin-sensitive sites in ovarian blood vessels.
肝脂酶被认为通过参与高密度脂蛋白的代谢在类固醇生成中发挥作用。我们研究了该酶的活性、合成、分布和摄取,并评估了其在黄体化卵巢中的mRNA含量。我们发现,在类固醇生成高峰期,用孕马血清促性腺激素-人绒毛膜促性腺激素处理的未成熟大鼠的卵巢含有可被肝素释放的肝脂酶样活性,该活性可被从大鼠肝脏肝素灌注液中分离出的肝脂酶纯化抗体以剂量依赖性方式中和。定量免疫电子显微镜显示,卵巢肝脂酶存在于内皮细胞中,在黄体血管中的含量比基质血管中的高3倍。然而,卵巢并不合成肝脂酶,因为在用[35S]甲硫氨酸孵育的分散黄体化颗粒细胞的培养基中未免疫分离到放射性标记的酶,而在对照细胞(肝细胞)的培养基中则存在。同样,通过Northern印迹或狭缝印迹分析或聚合酶链反应,在肝脏中可检测到肝脂酶mRNA,而在卵巢或肾脏中则未检测到。最后,经尾静脉注射的125I标记肝脂酶很快从体循环中清除,在肝脏、卵巢、肾脏和脾脏中蓄积。随后注射肝素导致放射性迅速重新出现在血流中,肝脏和卵巢中的放射性标记明显下降,但肾脏中的放射性标记略有下降,脾脏中则没有下降。外源性125I-牛血清白蛋白也在所有四个器官中蓄积,但随后给予肝素并未使其从肝脏或卵巢中排出。综上所述,这些数据表明,具有类固醇生成活性的卵巢拥有但不合成肝脂酶。相反,源自其他地方(可能是肝脏)的肝脂酶从循环中在卵巢血管中对肝素敏感的部位蓄积。