本文關(guān)鍵詞： 球狀脂聯(lián)素 卵巢 血管新生 血管內(nèi)皮細(xì)胞 AMPK　出處：《南方醫(yī)科大學(xué)學(xué)報》2017年05期 　論文類型：期刊論文

【摘要】：目的分離并鑒定小鼠卵巢微血管內(nèi)皮細(xì)胞,并利用卵巢微血管內(nèi)皮細(xì)胞研究球狀脂聯(lián)素對小鼠卵巢血管新生的影響。方法采用Percoll密度梯度離心法分離純化小鼠卵巢微血管內(nèi)皮細(xì)胞,并采用免疫熒光檢測細(xì)胞表面的卵泡刺激素受體、黃體生成素受體以及血管內(nèi)皮細(xì)胞標(biāo)記v WF;通過血管生成因子(VEGFA)處理檢測卵巢微血管內(nèi)皮細(xì)胞在Matrigel上的毛細(xì)管形成特性。重組的球狀脂聯(lián)素蛋白處理卵巢微血管內(nèi)皮細(xì)胞,通過MTS檢測細(xì)胞增殖;細(xì)胞劃痕愈合法檢測細(xì)胞遷移;Matrigel基質(zhì)膠的血管形成,Western blot檢測單磷酸腺苷活化蛋白激酶(AMPK)磷酸化激活。結(jié)果分離出的細(xì)胞表現(xiàn)為卵泡刺激素受體陰性,黃體生成素受體陽性,v WF陽性,且VEGFA能夠促進(jìn)血管生長,具有卵巢特異的血管內(nèi)皮生長特性。高劑量濃度(1μg/m L和3μg/m L)的重組的脂聯(lián)素球狀結(jié)構(gòu)域蛋白處理后卵巢微血管內(nèi)皮細(xì)胞數(shù)量分別增加(158.72±14.50)%和(186.50±4.20)%,促進(jìn)細(xì)胞增殖(P0.01);高劑量濃度(1μg/m L和3μg/m L)的重組的脂聯(lián)素球狀結(jié)構(gòu)域蛋白處理后卵巢微血管劃痕愈合率分別為(49.43±3.43)%(P0.05)和(69.67±1.2)%(P0.01);3μg/m L的脂聯(lián)素處理卵巢微血管內(nèi)皮細(xì)胞形成的毛細(xì)管狀長度為對照組的7.63±0.66倍(P0.01)。3μg/m L的球狀脂聯(lián)素蛋白處理饑餓處理后的卵巢微血管內(nèi)皮細(xì)胞培養(yǎng)15 min后p AMPK/AMPK、30 min后p AMPK/AMPK顯著高于未加入蛋白處理的對照(P0.01)。Compound C抑制了球狀脂聯(lián)素促進(jìn)的卵巢微血管內(nèi)皮細(xì)胞的管狀結(jié)構(gòu)形成及AMPK磷酸化激活。結(jié)論成功分離了卵巢微血管內(nèi)皮細(xì)胞,球狀脂聯(lián)素蛋白能夠通過激活A(yù)MPK信號途徑促進(jìn)卵巢微血管內(nèi)皮細(xì)胞的血管新生。
[Abstract]:Objective to isolate and identify mouse ovarian microvascular endothelial cells. The effects of globular adiponectin on ovarian angiogenesis in mice were studied by using ovarian microvascular endothelial cells. Methods Mouse ovarian microvascular endothelial cells were isolated and purified by Percoll density gradient centrifugation. The follicle stimulating hormone receptor, luteinizing hormone receptor and vascular endothelial cell labeled vWF were detected by immunofluorescence. The capillary formation characteristics of ovarian microvascular endothelial cells on Matrigel were detected by VEGF treatment. Recombinant spherical adiponectin was used to treat ovarian microvascular endothelial cells. Cell proliferation was detected by MTS. Cell migration was detected by scratch healing method. Angiogenesis of Matrigel matrix glue. The phosphorylation of adenosine monophosphate activated protein kinase (AMPK) was detected by Western blot. The results showed that the isolated cells were negative for follicle stimulating hormone receptor and positive for luteinizing hormone receptor. V WF was positive and VEGFA could promote blood vessel growth. Ovarian specific vascular endothelial growth characteristics. High dose concentration of 1 渭 g / mL and 3 渭 g / mL). The number of ovarian microvascular endothelial cells was increased by 158.72 鹵14.50% and 186.50 鹵4.20%, respectively. Promote cell proliferation (P0.01); The healing rate of ovarian microvascular scratches after treatment with 1 渭 g / mL and 3 渭 g / mL recombined adiponectin globular domain proteins was 49.43 鹵3.43% (P < 0.05). P0.05) and 69.67 鹵1.2% P0.01C; The capillary length of ovarian microvascular endothelial cells treated with 3 渭 g / mL adiponectin was 7.63 鹵0.66 times longer than that of control group (P 0.01). 3 渭 g / mL of globular adiponectin treated with starvation treated ovarian microvascular endothelial cells cultured for 15 min, p AMPK/AMPK. After 30 min, p AMPK/AMPK was significantly higher than that of control without protein treatment (P0.01). Compound C inhibited tubular structure formation and AMPK phosphorylation activation of ovarian microvascular endothelial cells stimulated by globular adiponectin. Conclusion OVECs were isolated successfully. Spherical adiponectin can promote angiogenesis of ovarian microvascular endothelial cells by activating AMPK signaling pathway.
【作者單位】： 暨南大學(xué)附屬第一醫(yī)院麻醉科;暨南大學(xué)再生醫(yī)學(xué)教育部重點實驗室;暨南大學(xué)生命科學(xué)技術(shù)學(xué)院發(fā)育與再生生物學(xué)系;
【基金】：國家自然科學(xué)基金(30900232) 中央高�；究蒲袠I(yè)務(wù)費專項資金(21610202) 天河區(qū)科技計劃項目(201404KW041) 廣東省優(yōu)秀博士學(xué)位論文作者資助項目(sybzzxm201033)~~
【分類號】：R614
【正文快照】： Effect of globular adiponectin on proliferation,migration and tube formation of ovarian mi-crovascular endothelial cellsCHEN Lei1,LU Xiaosheng2,3,LI Yalan1,MAO Zhoufei2,3,XIAO Luanjuan2,3,YU Yanhong2,3Department of Anesthesia,First Affiliated Hospital1,K

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