Analysis on tyrosinekinase receptor expression and tubal infertility

(研究一)分析兔输卵管结扎后30min至14d不同时间点输卵管的组织学和生理学变化。前3d输卵管血流量减少，输卵管积液中TUNEL阳性上皮细胞和NO含量增加。L-NAME注射液能降低NO水平和液体蓄积。而L-NAME使TUNEL阳性的平滑肌细胞增多。上述结果提示，兔输卵管结扎后NO水平的快速升高可能在输卵管组织损伤和重塑中起重要作用。(研究2)建立小鼠子宫缺血再灌注模型，观察子宫和输卵管的病理变化。缺血诱导子宫内膜和输卵管细胞凋亡。月经过多与人类月经期间子宫内膜相似的病理特征再灌流后子宫组织中肿瘤坏死因子α的表达水平升高。肿瘤坏死因子受体p55缺陷小鼠的细胞凋亡率减少，证实了肿瘤坏死因子α在损伤诱导细胞凋亡中的重要作用。我们的模型似乎适合于研究人类月经和输卵管性不孕。(研究3)采用小鼠子宫缺血再灌注模型，分析受精和胚胎发育情况。在该模型中，受精率和胚胎发育率都有所下降。而注射超氧化物歧化酶后则升高。这表明氧化应激是胚胎发育的重要因素。该模型可用于输卵管性不孕症的研究。(研究4)分析了小鼠正常输卵管和缺血输卵管的酪氨酸激酶受体图谱。利用简并引物RT-PCR，最具特征的受体是c-Met。C-met基因在小鼠输卵管缺血再灌流后的mRNA和免疫组织化学中表达。注射抗c-met抗体后，缺血后输卵管再生减少。提示c-met和肝细胞生长因子在输卵管再生中起重要作用。

(study1) We analyzed histological and physiological changes in the rabbit oviduct at various time-points from 30 min to 14 day after ligation of both ends of oviduct. In the first 3days, the blood flow decreased, and TUNEL positive epithelial cells and NO level in the oviduct fluid increased. L-NAME injection reduced NO level and the accumulation of fluid. However, L-NAME increased TUNEL positive smooth muscle cells. These results indicated that the rapid increase in NO level after ligation may play a major role in the tissue injury and remodeling in rabbit oviduct. (study2) We established a uterine ischemia-reperfusion model in mice to investigate the pathological changes in the uterus and oviduct. Ischemia induced apoptosis in endometrium and oviduct. The pathological features similar to those seen in the human endometrium during menstruation. The expression levels of TNF α mRNAs in uterus increased after reperfusion. Apoptosis decreased in TNF-R p55-deficient mice, confirming the important role of TNF α in injury-induced apoptosis. Our model seems suitable for investigating human menstruation and tubal infertility. (study3) We analyzed fertilization and embryonal development with mouse uterine ischemic reperfusion model. In this model, the fertilization rate and embryonal development decreased. However, it was increased after injection of superoxide dismutase. This suggest that oxidative stress is the important factor in embryonal development. This model is useful in study of tubal infertility. (study4) We analyzed tyrosine kinase receptor profiles of mouse normal oviduct and ischemic oviduct. Using degenerated primer RT-PCR, the most characteristic receptor was c-met. C-met was expressed in mouse oviduct after ischemia reperfusion in mRNA and immunohistochemistry. After injection of anti c-met antibody, oviduct regeneration after ischemia was decreased. This suggest that c-met and hepatocyte growth factor was important in the oviduct regeneration.