Mikulska D
Katedry i Zakładu Histologii i Embriologii Pomorskiej Akademii Medycznej w Szczecinie.
Ann Acad Med Stetin. 1995;41:131-43.
It is apparent from numerous communications that hysterosalpinogography (HSG), besides its diagnostic value, may also claim therapeutic activity. After such examinations, more frequent pregnancies were recorded in previously infertile women without resorting to supplementary treatment. Pregnancies were more numerous when applying oil-based contrast media than water-based contrast ones. Hypothesis has been put forward as to the possibility of modulating activity of these agents exerted upon the peritoneal and oviductal macrophages. Increased amount of these cells as well as excessive phagocytosis of spermatozoa was shown in case of endometriosis, one of the most frequent causes of infertility. It is supposed that excessive phagocytosis of spermatozoa by peritoneal macrophages (PM) may be the cause of infertility. Studies were performed to determine the effect of oil- and water-soluble contrast media, on sperm phagocytosis by PM in vitro. PM were obtained from Wistar female rats. Subsequently, the cells were centrifuged, washed and suspended in culture medium. Next 1 million PM were allowed to attach to the cover glass for one hour under standard conditions of incubation. Dilutions of 0.25%, 0.5%, 1.0% Lipiodol Ultra--Fluid (Byk Gulden Konstanz) and Uropolinum 60% (Polfa) were added to chambers with incubated PM for next one hour. The controls were cultured in the same conditions without contrast medium added. Sperm were isolated from the epididymal cauda of male rats and subsequently suspended in culture medium at a concentration 1 million/ml and added to equivalent volume of cultured macrophages for 1.5 hours. After the exposure time the cultures were washed and stained. The spermiophagic index (SPI) was determined. SPI = number of phagocytosed sperm cells/number of macrophages x 100. Statistical analysis was performed by means of Student t-test. Additional histochemical reactions were made and scanning as well as transmission electron microscopy studies were accomplished. The established results (Fig. 1-9) indicate that lipiodol inhibits phagocytosis of spermatozoa by PM stronger (1% lipodol SPI = 1.99 + 0.94 < 0.0001) than uropolinum (1% uropolinum SPI = 5.07 +/- 1.02 p < 0.0001). In control studies, without contrast medium added, SPI was equal to 14.66 +/- 3.12 (p < 0.00001). Marked inhibition involving phagocytosis of spermatozoa was detected already after the treatment with lipiodol in low concentration (0.25% lipiodol SPI = 3.73 +/- 0.89 p < 0.0001). It failed to be so distinct after treating macrophages with uropolinum in low concentration (0.25% uropolinum SPI = 8.34 +/- 1.50 p < 0.0001). Scanning electron microscopy studies have disclosed that in the cultures of macrophages with spermatozoa, the macrophages spread well over the glass bottom. The macrophages' cytoplasmic membrane was highly folded with numerous protrusions different in size and shape. They covered the digested parts of spermatozoon causing its fragmentation. In cultures of macrophages with spermatozoa and 1% uropolinum, the structure of macrophages was similar to that in the control group. Accumulations and conglomerations of a large number of macrophages with spermatozoa were additionally revealed. In cultures of macrophages with spermatozoa and 1% lipiodol, the macrophages were not spread, but rather spherical. Also the cytoplasmic membrane protrusions were small, short, fine and appeared in smaller number. Occasionally areas of smooth cytoplasmic membrane without any folds were seen. Such macrophages did not phagocytose sperm. Transmission electron microscopy studies of macrophages cultured with sperm have disclosed that the macrophages' cytoplasmic membrane was highly folded with numerous protrusions different in size and shape. The latter encircled and closed parts of sperm (head, midpiece or tail) inside inside endosomes, finally linking the latter with lysosomes. The ultrastructure of macrophages cultured with spermatozoa and 1% uropolinum was s
从大量交流中可以明显看出，子宫输卵管造影术（HSG）除了具有诊断价值外，还可能具有治疗作用。在进行此类检查后，以前不孕的女性在未采取辅助治疗的情况下受孕更为频繁。使用油基造影剂时的受孕次数比使用水基造影剂时更多。有人提出了关于调节这些药物对腹膜和输卵管巨噬细胞作用活性的可能性的假设。子宫内膜异位症是最常见的不孕原因之一，在这种情况下，这些细胞的数量增加以及精子的过度吞噬现象较为明显。据推测，腹膜巨噬细胞（PM）对精子的过度吞噬可能是不孕的原因。进行了研究以确定油溶性和水溶性造影剂对体外PM吞噬精子的影响。PM取自Wistar雌性大鼠。随后，将细胞离心、洗涤并悬浮于培养基中。接下来，在标准培养条件下，让100万个PM附着在盖玻片上1小时。将0.25%、0.5%、1.0%的超液态碘油（拜耳先灵康斯坦茨公司）和60%的泛影葡胺（波法公司）稀释液加入培养有PM的培养室中，再培养1小时。对照组在相同条件下培养，但不添加造影剂。从雄性大鼠附睾尾部分离精子，随后将其以100万/ml 的浓度悬浮于培养基中，并加入等量体积的培养巨噬细胞中，共培养1.5小时。暴露时间结束后，对培养物进行洗涤和染色。测定精子吞噬指数（SPI）。SPI = 吞噬精子细胞数/巨噬细胞数×100。采用学生t检验进行统计分析。还进行了额外的组织化学反应，并完成了扫描电子显微镜和透射电子显微镜研究。已得出的结果（图1 - 9）表明，碘油比泛影葡胺更能强烈抑制PM对精子的吞噬作用（1%碘油SPI = 1.99 + 0.94 < 0.0001，1%泛影葡胺SPI = 5.07 +/- 1.02 p < 0.0001）。在对照研究中，不添加造影剂时，SPI等于14.66 +/- 3.12（p < 0.00001）。在低浓度碘油处理后（0.25%碘油SPI = 3.73 +/- 0.89 p < 0.0001）就已检测到对精子吞噬的明显抑制。而低浓度泛影葡胺处理巨噬细胞后，这种抑制并不那么明显（0.25%泛影葡胺SPI = 8.34 +/- 1.50 p < 0.0001）。扫描电子显微镜研究表明，在含有精子的巨噬细胞培养物中，巨噬细胞在玻璃底部铺展良好。巨噬细胞的细胞质膜高度折叠，有许多大小和形状各异的突起。它们覆盖精子的消化部分，导致精子碎片化。在含有精子和1%泛影葡胺的巨噬细胞培养物中，巨噬细胞的结构与对照组相似。还额外发现大量含有精子的巨噬细胞聚集和成团现象。在含有精子和1%碘油的巨噬细胞培养物中，巨噬细胞没有铺展，而是呈球形。细胞质膜突起也小、短、细且数量较少。偶尔可见没有任何褶皱的光滑细胞质膜区域。这样的巨噬细胞不吞噬精子。对与精子一起培养的巨噬细胞进行透射电子显微镜研究表明，巨噬细胞的细胞质膜高度折叠，有许多大小和形状各异的突起。这些突起在内体内部环绕并封闭精子的部分（头部中段或尾部），最终将其与溶酶体连接起来。与精子和1%泛影葡胺一起培养的巨噬细胞的超微结构……
