天然产物研究与开发 ›› 2018, Vol. 30 ›› Issue (5): 767-772.doi: 10.16333/j.1001-6880.2018.5.007

• 研究论文 • 上一篇    下一篇

齐墩果酸对自然衰老大鼠睾丸DNA损伤保护作用及机制研究

马娜1,张长城1,陈茜1,刘珍财1,韩贵芳1,刘静1,袁丁2,赵海霞*   

  1. 1三峡大学医学院,宜昌 443002;2三峡大学仁和医院,宜昌 443002
  • 出版日期:2018-06-04 发布日期:2018-06-04
  • 基金资助:

    国家自然科学基金(81503334;81573931;81774316;81373881);三峡大学学位论文培优基金项目资助(2018SSPY095)

Protective Effects and Mechanism of Oleanolic Acid on Testicular DNA Damage in Natural Aging Rats

MA Na1, ZHANG Chang-cheng1, CHEN Qian1, LIU Zhen-cai1, HAN Gui-fang1, LIU Jing1, YUAN Ding2, ZHAO Hai-xia1*   

  1. 1Medical College of China Three Gorges University,Yichang 443002,China; 2Ren-He Hospital of China Three Gorges University,Yichang 443001,China
  • Online:2018-06-04 Published:2018-06-04

摘要: 探讨齐墩果酸(Oleanolic Acid,OA)对自然衰老大鼠睾丸DNA损伤的保护作用及其可能机制。将30只SPF级18月龄SD大鼠随机分为三组:自然衰老组、OA低、高剂量组,每组10只,另取10只3月龄大鼠作为青年对照组。OA低、高剂量组分别灌胃给予OA 5、25 mg/kg,自然衰老组和青年对照组给予生理盐水,每周灌胃6天,周日不给药,连续给药6个月。末次给药后禁食不禁水12 h,称重,麻醉后处死大鼠,快速取出睾丸组织。HE染色观察睾丸组织形态变化,免疫组织化学法检测DNA损伤相关蛋白53BP1、ATR的表达,Western Blot检测睾丸组织中衰老相关蛋白p21和DNA损伤相关蛋白γ-H2AX、Chk1、p-p53的表达水平。HE结果表明,OA能显著改善增龄所致的睾丸组织形态变化;与青年对照组相比,自然衰老组睾丸组织DNA损伤标志蛋白γ-H2AX表达水平明显升高,而给予OA干预后其表达显著降低;此外,与青年对照组比较,自然衰老组睾丸组织中衰老相关蛋白p21的表达显著增高,DNA损伤相关蛋白53BP1、ATR的阳性灶点数及其下游Chk1、p-p53的蛋白表达明显升高,OA低、高剂量组均能显著降低上述蛋白的表达。以上结果表明OA对自然衰老大鼠睾丸组织DNA损伤有较好的保护作用,机制与调节ATR/Chk1/P53信号通路有关。

关键词: 齐墩果酸, 衰老, 睾丸, DNA损伤

Abstract: To study the protective effects of Oleanolic Acid on Testicular DNA Damage in natural aging rats,and to explore the possible mechanism.The 18-month old male SD rats were randomly divided into natural aging group,OA low and high dose group,with 10 rats in each group.Another 10 3-month old rats were used as young control group.OA low and high dose groups were given gastric gavage of OA 5 and 25 mg/kg,respectively,while the young control group and the natural aging group were given saline six times per week,lasting for 6 months.After fasting for 12 hours,the rats were sacrificed.Then,the testes were immediately removed and weighed.The change of testicular tissue morphology was observed by HE staining.The expression and localization of DNA damage-associated protein 53BP1 and ATR were observed by Immunohistochemistry.The relative expression levels of senescence associated protein p21 and DNA damage-related proteins γ-H2AX,Chk1 and p-p53 in testes were detected by Western Blot.HE showed that OA significantly improved the testicular tissue structure of natural aging rats.Compared with young control group,the levels of γ-H2AX protein in testis were significantly upregulated in natural aging group.However,the expression of γ-H2AX significantly decreased in OA-treated groups.In addition,the numbers of ATR-and 53BP1-positive cells,and the protein expression levels of p21,Chk1,and p-p53 were significantly upregulated in testis of rats in natural aging group when compared with young control group.As expected,OA significantly restored these changes.Taken together,OA protected against testicular DNA damage of natural aging rats,which may be associated with ATR/Chk1/P53 signaling.

Key words: Oleanolic acid, Aging, Testis, DNA damage

中图分类号: 

R961