摘要
目的:探讨携带Bcl-2基因的慢病毒局部注射修复用环磷酰胺(CTX)损伤卵巢结构功能的作用。方法:将Wistar大鼠分为空白对照组、CTX组、Bcl-2组、EGFP组、空载体+CTX组、实验组(注射pGC-FU-Bcl-2+CTX)6个组,每组16只;将纯化的慢病毒悬液经开腹注射入双侧卵巢内,CTX、空载体+CTX及实验组动物采用CTX经腹腔注射的方式建立POF动物模型。注射结束后1、15、30、45天及60天留取各组血标本,并于此5个时间点分批处死大鼠留取卵巢。观察绿色荧光及其分布以及卵巢结构的变化情况,阴道脱落细胞涂片观察大鼠动情周期的变化,以放射免疫法检测雌二醇(E2)及卵泡刺激素(FSH)浓度。结果:卵巢内直接注射慢病毒后,可见卵巢间质组织及卵泡壁上出现明亮的绿色荧光,卵泡内见不到绿色荧光。空载+CTX组及CTX组可见皮质和髓质组织中细胞明显受损,CTX注射结束后15天实验组大鼠皮质和髓质细胞形态基本正常,血清E2浓度比其余各化疗组显著升高,FSH浓度升高不明显,与正常大鼠的激素水平仍有显著性差异。结论:携带Bcl-2基因的慢病毒可防护CTX对卵巢组织结构的损伤,部分改善卵巢功能,Bcl-2基因对CTX诱导的大鼠卵巢损伤有一定的保护作用。
Objective： To explore the effect of lentivirus - mediated Bcl - 2 gene for repair of ovarian structural and functional in- jury induced by eyclophosphamide （ CTX ） in rats. Methods ： Wistar rats were divided into 6 groups ： blank control group, CTX group, Bcl - 2 group, EGFP group, empty vector ＋ CTX group, experimental group （receiving injection of pGC - FU - Bcl - 2 ＋ CTX） , 16 rats in each group; the purified lentivirus suspension was injected into bilateral ovaries transabdominally, premature ovarian failure （POF） models of rats in CTX group, empty vector ＋ CTX group, and experimental group were established by transabdominal injection of CTX, blood sam- ples were obtained at 1, 15, 30, 45, and 60 days, and ovarian samples were obtained at the above - mentioned rice time points after killing the rats. The distribution of green fluorescence and the changes of ovarian structure were observed, the change of oestrous cycles was ob- served by vaginal exfoliative cells smears, RIA was used to detect the contents of estradiol and follicle - stimulating hormone （FSH） . Re- sults： Light green fluorescence was found in interstitial tissue and follicular wall, and it was not observed in follicle; obvious injury of cells in ovarian cortex and medulla in empty vector ＋ CTX group and CTX group was observed, cell morphology of ovarian cortex and medulla cells at 15 days were basically normal, the serum estradiol level were significantly higher than those in the other chemotherapy groups, the increase of FSH was not obvious, compared with normal rats, there was statistically significant difference. Conclusion ： Lentivirus - mediated Bcl - 2 gene can prevent injury of CTX to ovarian structure, and improve ovarian function partially, Bcl -2 gene has a certain protective effect on CTX - induced ovarian injury of rats.
出处
《中国妇幼保健》
CAS
北大核心
2013年第1期143-147,共5页
Maternal and Child Health Care of China
基金
2010年国家自然基金项目〔81041101〕
2010年广东省自然科学基金项目〔10451051501004704〕
