【摘要】 目的探讨哈蟆油(OR)对D-半乳糖所致雌性衰老大鼠子宫组织细胞增殖负性调控因子p16和正性调控因子cyclinD1蛋白表达的影响,进一步探讨OR延缓雌性大鼠生殖器官衰老机制。方法 SPF级SD雌性青年大鼠40只随机分为模型组(D-gal组)、维生素E(VE组)、哈蟆油高剂量组(OR-H组)、中剂量组(OR-M)、低剂量组(OR-L组),每组8只,D-半乳糖颈背部皮下注射42 d,建立亚急性衰老模型。另取雌性青年大鼠8只,同样部位每日注射生理盐水,作为空白组。第15天开始灌胃给药,给药时间28 d。给药结束后,免疫组化法检测衰老大鼠子宫组织p16和免疫印迹法检测cyclinD1蛋白的表达情况。结果雌性衰老大鼠子宫组织免疫组化结果表明,子宫组织p16阳性染色多为胞浆着色,见于子宫内膜、上皮细胞胞浆、子宫间质腺体腺上皮细胞胞浆,外膜上皮也有表达,弥漫性、灶性分布均有。D-gal组p16阳性细胞积分与空白组比较升高(P<0.01)。OR各剂量组与D-gal组相比,p16阳性细胞积分降低,差异有显著性(P均<0.01)。免疫印迹法结果表明D-gal组子宫组织cyclinD1蛋白表达与空白组比较降低,差异有显著性(P<0.01)。OR-H、M、L组cyclin D1蛋白表达与D-gal组比较,表达均升高(P值均<0.01),OR-H组尤为明显。结论应用OR可以减缓衰老子宫组织结构的损伤,改善衰老子宫萎缩和衰老程度,OR可降低雌性衰老大鼠子宫组织细胞增殖负性调控因子p16蛋白的高表达,同时显著提高细胞增殖正性调控因子cyclinD1蛋白的表达,促进衰老雌性大鼠子宫细胞增殖。哈蟆油延缓雌性生殖器官衰老作用可能通过调控子宫p16-cyclinD1信号通路来促进有关增殖调控蛋白表达发挥延缓衰老作用。更多还原

【Abstract】 Objective To investigate the expressions of uterus tissue cellular proliferation negativity regulating factor p16 and positive regulating factor cyclinD1 in aged female rats with Oviductus Ranae(OR) by D-galactose.Methods 40 young female SD rats(SPF grade) were randomly divided into 5 groups: the OR-H,OR-M,OR-L,Vitamin E and D-gal groups with 8 in each group.The 40 rats received subcutaneous injection of D-gal on the nape for 42 days to establish aging model.Another 8 rats were injected daily with normal saline(NS) at the same position as the normal control group.From the 15th day of the experiment,the rats were given oral OR(in OR group) or Vitamin E(in VE group) for 28 days.Then all the rats were sacrificed.p16 in uterus tissue was detected by immuno-histochemistry(IHC),and cyclinD1 was detected by Western-blotting method.Results The positive staining of the p16 protein in uterus tissue was mainly located in cytoplasm,which were on the endometrium,the cytoplasm of epithelial cells and interstitial glands and a few in uterine glandular epithelium,and their distribution was diffuse and focus.Positive cells integral of p16 in D-gal female aged rats was significantly increased than that of normal control group(P<0.01).After taking OR,the integral of p16-positive cell in all OR groups was significantly decreased than that of D-gal group(P<0.01).Expression of cyclinD1 protein in uterus tissue of D-gal female rats was decreased than that of normal control group and the difference was significant(P<0.01).The expression of cyclinD1 in all OR groups was significantly increased compared with D-gal group(P<0.01).The expression of cyclinD1 in OR-H group was the highest in all OR groups.Conclusions OR could restore impairment of pathology and maintain cellular normal form,alleviate the pathological change of uterus.OR could decrease the over-expression of p16 protein in uterus and promote uterus cell proliferation of aging female rats.Meanwhile,OR could enhance expression of cyclinD1 in uterus of female aged rats.The possible anti-aging effects of OR is producing regulation of p16,cyclinD1 signaling pathways to promote expression of the proliferation regulatory protein and the signaling pathways,which plays an important role in delaying aging.更多还原