医学免费论文:丹参对冻融小鼠卵巢同种异体移植后卵泡存活率的影响及其作用机制
【摘要】 目的 :探讨丹参注射液对冻融小鼠卵巢同种异体移植缺血低氧损伤的保护作用及可能机制。方法:收集B6C2F1第1天新生(D1)小鼠卵巢,慢冻速融后移植至B6C2F1代8~12周雄鼠的肾被膜下,移植后小鼠按给药不同分为对照组和丹参组,分别于移植后1 d(24 h)、2 d(48 h)和14 d回收移植物,进行超氧化物歧化酶(SOD)及诱导型一氧化氮合酶(iNOS)的RT-PCR半定量测定,同时移植后14 d回收移植物做HE染色全卵巢卵泡计数及PCNA免疫组化分析。结果:移植后各时间段卵巢组织SOD mRNA的表达,丹参组均高于对照组,而iNOS mRNA的表达则均低于对照组。移植后14 d回收全卵巢总卵泡数,丹参组(2 709±234)多于对照组(1 889±202),P<0.05;卵泡存活率,丹参组(64.1%)显著高于对照组(44.7%),P<0.05。结论 :丹参注射液可能通过上调SOD mRNA的表达,抑制iNOS mRNA的表达而改善冻融小鼠卵巢同种异体移植缺血低氧性损伤,从而提高卵泡的存活率。
【关键词】 丹参;卵巢;冷冻保存;移植
Effects of danshen injection on the survival rates of follicle in heterotopically grafted newborn mouse ovaries and the relative mechanism LIN Hai-yan, XIONG Cheng-rong, ZHU Qin, WU Dan-dan,TANG Fei,ZHOU Ying. Department of Reproduction Center,the First Affiliated Hospital of Wenzhou Medical College, Wenzhou,325000医.学.全.在.线www.lindalemus.com
Abstract: Objective: To explore the protective effects of danshen injection on ischemia injury in heterotopically grafted newborn mouse ovaries. Methods:Ovaries from B6C2F1 newborn mice were cryopreservation and the thawed ovaries were transplantated into kidney capsules of 8~12 w adult male mice.The mice were divided into two groups:control group and danshen group.The recipient mice were sacrificed on 1 d,2 d and 14 d respectively after transplantation. The grafts from 1 d, 2 d and 14 d were removed to assess the mRNA expressions of SOD and iNOS with RT-PCR. Meanwhile,the grafts from 14 d were processed histologically for follicle counting and immunohistochemically for PCNA. Results: SOD-mRNA expression of ovarian tissue in the duration period after transplantation were higher than that of danshen group, and the iNOS-mRNA expression were lower than that of control group. Number of follicles in 2 week old grafts after transplan-tation of danshen group (2 709±234) was more than that of control group (1 889±202).There was an significant improvement in survival rate from 44.7% to 64.1% if host animals were to be administered with danshen injection. Conclusion: Danshen injection may improved the survival rate of ovaries by reducing ischeamia injury in heterotopically grafted newborn mouse ovaries with the increase of SOD-mRNA expression and the decrease of iNOS-mRNA expression.
Key words: danshen;ovary;cryopreservation;transplantation
半个世纪以来的研究表明,卵巢组织的冷冻保存和卵巢移植是目前保留癌症患者生育能力的较好方法[1]。移植后缺血低氧损伤导致的大量卵泡丢失仍然是阻碍移植卵巢组织功能恢复的关键问题[2],其机制可能是移植物中脂质过氧化和其他氧自由基所造成的缺血低氧损伤[3]。丹参是一种常用的活血化瘀中药,它能清除自由基、改善血流变化和微循环状态。许多研究表明,丹参对心、肝、肺等多种组织器官的缺血损伤具有明显的改善作用,但对于冻融卵巢组织移植过程中缺血低氧导致卵泡损伤的保护作用尚未见报道。本实验观察丹参注射液对冻融小鼠卵巢移植早期缺血低氧卵泡损伤的保护作用及相关基因超氧化物歧化酶(SOD)和诱导型一氧化氮合酶(iNOS)的表达影响,探讨丹参的外源性保护作用及其与移植物低氧相关基因表达之间的关系。为进一步探讨如何降低冻融卵巢组织移植早期缺血低氧损伤机制提供实验依据。