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醫(yī)學(xué)免費論文:丹參對凍融小鼠卵巢同種異體移植后卵泡存活率的影響及其作用機制
【摘要】 目的 ：探討丹參注射液對凍融小鼠卵巢同種異體移植缺血低氧損傷的保護作用及可能機制。方法：收集B6C2F1第1天新生(D1)小鼠卵巢，慢凍速融后移植至B6C2F1代8～12周雄鼠的腎被膜下，移植后小鼠按給藥不同分為對照組和丹參組，分別于移植后1 d（24 h)、2 d(48 h)和14 d回收移植物，進行超氧化物歧化酶（SOD)及誘導(dǎo)型一氧化氮合酶（iNOS)的RT-PCR半定量測定，同時移植后14 d回收移植物做HE染色全卵巢卵泡計數(shù)及PCNA免疫組化分析。結(jié)果：移植后各時間段卵巢組織SOD mRNA的表達，丹參組均高于對照組，而iNOS mRNA的表達則均低于對照組。移植后14 d回收全卵巢總卵泡數(shù)，丹參組（2 709±234)多于對照組（1 889±202)，P＜0.05；卵泡存活率，丹參組（64.1%)顯著高于對照組（44.7%)，P＜0.05。結(jié)論 ：丹參注射液可能通過上調(diào)SOD mRNA的表達，抑制iNOS mRNA的表達而改善凍融小鼠卵巢同種異體移植缺血低氧性損傷，從而提高卵泡的存活率。
【關(guān)鍵詞】 丹參；卵巢；冷凍保存；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醫(yī).學(xué).全.在.線m.gydjdsj.org.cn
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
半個世紀(jì)以來的研究表明，卵巢組織的冷凍保存和卵巢移植是目前保留癌癥患者生育能力的較好方法[1]。移植后缺血低氧損傷導(dǎo)致的大量卵泡丟失仍然是阻礙移植卵巢組織功能恢復(fù)的關(guān)鍵問題[2]，其機制可能是移植物中脂質(zhì)過氧化和其他氧自由基所造成的缺血低氧損傷[3]。丹參是一種常用的活血化瘀中藥，它能清除自由基、改善血流變化和微循環(huán)狀態(tài)。許多研究表明，丹參對心、肝、肺等多種組織器官的缺血損傷具有明顯的改善作用，但對于凍融卵巢組織移植過程中缺血低氧導(dǎo)致卵泡損傷的保護作用尚未見報道。本實驗觀察丹參注射液對凍融小鼠卵巢移植早期缺血低氧卵泡損傷的保護作用及相關(guān)基因超氧化物歧化酶（SOD)和誘導(dǎo)型一氧化氮合酶（iNOS)的表達影響，探討丹參的外源性保護作用及其與移植物低氧相關(guān)基因表達之間的關(guān)系。為進一步探討如何降低凍融卵巢組織移植早期缺血低氧損傷機制提供實驗依據(jù)。
