Huang J, Roby K F, Pace J L, Russell S W, Hunt J S
Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City 66160-7400.
J Leukoc Biol. 1995 Jan;57(1):27-35. doi: 10.1002/jlb.57.1.27.
Nitric oxide (NO), a potent and versatile free radical, is synthesized in macrophages and mast cells as well as in other types of cells by the inducible form of nitric oxide synthase (iNOS). In this study, cells containing iNOS were identified in the uteri of cycling mice by using a rabbit antibody generated to an iNOS-specific peptide. Macrophages were identified in semiserial sections of the same tissues with the monoclonal antibody, F4/80, and mast cells were identified by toluidine blue staining. In tissue sections of uteri obtained from mice in the four stages of the estrous cycle (8 to 11 mice per stage), iNOS immunoreactivity was strongest in diestrus-I uteri and weakest in diestrus-II uteri. Myometrial mast cells and endometrial epithelial cells were prominent locations of iNOS, and specific protein was also present in myometrial smooth muscle and macrophage-like cells in the endometrial stroma. Because cyclic variations suggested regulation of iNOS expression by ovarian steroid hormones, studies were done using ovariectomized mice. Seven days after ovariectomy, immunoreactive iNOS was low but detectable in mast cells and luminal epithelial cells. In the uteri of ovariectomized, estradiol-17 beta (E2)-treated mice, mast cells were iNOS+ after 24 h whereas epithelial cells were negative; the reverse was observed in progesterone (P4)-treated mice. Both mast cells and epithelial cells were iNOS+ in the uteri of mice that had received a combination of E2 + P4. These results indicate that several types of uterine cells produce iNOS and that expression of this enzyme in specific cell lineages is governed by ovarian steroid hormones. The data are consistent with the postulate that NO derived from uterine leukocytes and other types of cells plays a role in uterine cyclicity and preparation for pregnancy.
一氧化氮（NO）是一种强效且多功能的自由基，可由诱导型一氧化氮合酶（iNOS）在巨噬细胞、肥大细胞以及其他类型的细胞中合成。在本研究中，通过使用针对iNOS特异性肽产生的兔抗体，在处于发情周期的小鼠子宫中鉴定出了含有iNOS的细胞。使用单克隆抗体F4/80在相同组织的半连续切片中鉴定巨噬细胞，并用甲苯胺蓝染色鉴定肥大细胞。在从发情周期四个阶段的小鼠获得的子宫组织切片中（每个阶段8至11只小鼠），iNOS免疫反应性在动情间期I子宫中最强，在动情间期II子宫中最弱。子宫肌层肥大细胞和子宫内膜上皮细胞是iNOS的主要定位部位，并且在子宫肌层平滑肌和子宫内膜基质中的巨噬细胞样细胞中也存在特异性蛋白。由于周期性变化提示iNOS表达受卵巢甾体激素调节，因此使用去卵巢小鼠进行了研究。去卵巢7天后，肥大细胞和腔上皮细胞中的免疫反应性iNOS较低但可检测到。在去卵巢并用雌二醇-17β（E2）处理的小鼠子宫中，24小时后肥大细胞为iNOS阳性而上皮细胞为阴性；在孕激素（P4）处理的小鼠中观察到相反的情况。在接受E2 + P4联合处理的小鼠子宫中，肥大细胞和上皮细胞均为iNOS阳性。这些结果表明，几种类型的子宫细胞可产生iNOS，并且该酶在特定细胞谱系中的表达受卵巢甾体激素控制。这些数据与以下假设一致，即源自子宫白细胞和其他类型细胞的NO在子宫周期性和妊娠准备中起作用。
