【作者】 张琳；

【导师】 杨静；

【作者基本信息】 武汉大学 ， 药理学， 2005， 硕士

【摘要】 慢性子宫颈炎（宫颈糜烂）是女性生殖系统的一种难愈性炎症,目前对于宫颈糜烂的防治尚无安全有效的方法,开发防治宫颈糜烂的新药具有重要意义。本文研究了重组人表皮生长因子（recombinant human epidermal growth factor,rhEGF）对实验性大鼠子宫颈炎的治疗作用,并对其机制进行了探讨。 一、以苯酚胶浆诱发大鼠子宫颈炎模型,♀SD大鼠随机分为6组:正常对照组,模型组,阳性对照组(聚甲酚磺醛栓剂5000.0μg·kg^-1),rhEGF给药组(2.0、4.0和8.0μg·kg^-1)。采用阴道给药,1粒/次,1次/d,共12d,模型组给予空白基质栓剂。末次给药后d2处死动物,观察宫颈组织病理形态学的改变,计算机图像分析系统分析炎症细胞数量,放射免疫分析法测定宫颈组织IL-8和PGE₂含量。研究结果表明rhEGF 2.0、4.0和8.0μg·kg^-1组和阳性对照组较模型组子宫颈组织病理学评分、炎性细胞数量、IL-8和PGE₂含量均显著降低,以8.0μg·kg^-1组作用最强。 二、研究重组人表皮生长因子（rhEGF）对大鼠宫颈鳞状上皮细胞（rat cervical epithelial cells,RCEC）体外增殖的影响,采用MTT法检测细胞增殖;流式细胞术（flow cytometry,FCM）分析细胞周期;逆转录聚合酶链反应（reverse transcription polymerase chain reaction,RT-PCR）检测Cyclin D1 mRNA表达水平。研究结果表明rhEGF 0.1,1.0,10和100μg·L^-1可不同程度刺激RCEC的增殖（刺激强度分别可达36.1%,41.3%,56.8%和48.4%）,且呈时间依赖性,以rhEGF 10μg·L^-1于d3的作用最强。rhEGF 0.1～100μg·L^-1明显提高S期细胞分数和Cyclin D1 mRNA表达,以10μg·L^-1于d3的作用最强,最高分别可达48.1%和50.7%。 本研究结果表明重组人表皮生长因子栓剂对实验性大鼠子宫颈炎有一定的治疗作用,其机制可能与下调IL-8和PGE₂介导的炎症反应有关。体外研究结果亦表明,rhEGF对大鼠宫颈鳞状上皮细胞（RCEC）有促进增殖的更多还原

【Abstract】 Chronic cervicitis(cervical erosion) is a kind of gynecic genital disease which lacks effective therapeutic strategies until now. This study investigated the therapeutic effect of recombinant human epidermal growth factor(rhEGF) on experimental cervicitis rats and related mechanisms both in vivo and in vitro.1. An experimental model of cervicitis in rats was established by application of phenol to the distal vaginal lumen. SD rats were randomly divided into 6 groups: normal control group, model group, positive control group and rhEGF 2.0 4.0 and 8.0 μg·kg~-1 treatment group. rhEGF suppositories or control were administered once daily for 12 days respectively. The mucosal injury were characterized by using a way of microscopic damage score; The inflammatory cell numbers were detected by using computer imaging analysis; The IL-8 and PGE2 content in cervical specimens were determined by radioimmunoassay. The pathological score, inflammatory cell numbers, IL-8 and PGE2 contents in rhEGF 2.0-. 4.0 and 8.0 μg·kg~-1 groups significantly decreased than that in the model group, the effect of rhEGF 8.0μg·kg~-1 was the most strong.2. Rat cervical epithelial cells(RCEC) were isolated from rat and cultured, the cell purity was evaluated by immunohistochemical technique; the proliferation of RCEC was detected by MTT assay. Cell cycle distributions were measured by flow cytometric assay(FCM). The Cyclin Dl mRNA expression was detected by reverse transcription polymerase chain reaction(RT-PCR). MTT assay showed that rhEGF 0.1, 1.0, 10,100 μg·kg~-1 stimulated increase in RCEC by 36.1%, 41.3%, 56.8%, 48.4% respectively, and the time course revealed that the accelerative effect reached the maximum at the concentration of 10 μg·kg~-1 on d 3. rhEGF (0.1~ 10 μg·kg~-1) increased the S phase cell fraction and cyclin Dl mRNA expression in concentration-dependent manner compared with the control, and the maximum accelerative effect was 48.10 % and 50.70%respectively.The results of this study showed that rhEGF 2.0^8.0 ug?kg~’ play an active role in the treatment of experimental cervicitis rats, which may be related to downregulating the inflammatory reaction mediated by IL-8 and PGE2. Furthermore, the study in vitro also proved that rhEGF promoted RCEC proliferation in the dose-dependant and time-dependant manner, possibly by accelerating the cell progression into S phase in the cell cycle resulting from upregulating Cyclin Dl mRNA expression.更多还原