Cepharanthine induces autophagy via PI3K/AKT/mTOR signaling pathway in ovarian cancer SKOV3 cells     

XIANG Fu-ying  JIANG Sheng-lin  CHENG Xian-ying 《园艺学报》2019,35(5):940

AIM: To investigate the autophagy of human ovarian cancer SKOV3 cells induced by cepharanthine and to explore its mechanism. METHODS: The effect of cepharanthine on the viability of ovarian cancer SKOV3 cells was measured by CCK-8 assay. The SKOV3 cells were treated with cepharanthine, and then the formation of autophagosome was observed with acridine orange staining under fluorescence microscope. The protein levels of LC3, AKT, p-AKT, mTOR, p-mTOR and GAPDH in the SKOV3 cells treated with cepharanthine were determined by Western blot.RESULTS: Cepharanthine significantly inhibited the viability of ovarian cancer SKOV3 cells in a dose-dependent manner (P<0.05). The number of the intracellular acidic autophagosomes with bright red fluorescence was significantly increased after cepharanthine treatment in the SKOV3 cells. The expression of LC3-Ⅱ in SKOV3 cells was significantly enhanced after cepharanthine treatment. Furthermore, treatment with cepharanthine in the SKOV3 cells also resulted in a significant down-regulation of phosphorylated form of AKT and mTOR (P<0.01), while the total protein level was not changed. Combination of cepharanthine and 3-methyladenine resulted in a substantial decrease in the cell viability compared with using cepharanthine alone.CONCLUSION: Cepharanthine significantly inhibits the growth of human ovarian cancer SKOV3 cells and induces the autophagy, which may be correlated with down-regulation of PI3K/AKT/mTOR signaling pathway.  相似文献   

Follicullar Development and Embryo Recovery Following 3 versus 8 FSH Injections in Heifers     

B. Purwantara  M. Schmidt  H. Callesen  T. Greve 《Acta veterinaria Scandinavica》1994,35(1):89

Ovarian follicular dynamics and embryo yield were studied during 2 different FSH regimens for superovulation of cattle. Twenty heifers were given intramuscular injections of FSH (total of 35 mg NIH) either once daily for 3 days (Group 3×1) or twice daily for 4 days (Group 4×2). At 72 h after the first FSH injection, each animal was injected with 0.75 mg cloprostenol. Inseminations were performed at 12 h and 24 h after the onset of heat. Transrectal ultrasonography was performed on the day of the first FSH injection, the day of cloprostenol injection, the day of insemination and finally on the day of embryo recovery (day 6 or 7 after heat). The numbers of small (2–4 mm), medium (5–9 mm) and large (>10 mm) size follicles were recorded. The total number of corpora lutea, eggs and transferable embryos were recorded on the day of embryo recovery. No differences were found between the 2 groups in either of the parameters studied (p>0.05). It can be concluded that treatment with this FSH preparation once daily for 3 days gives a folliculogenic and superovulatory response similar to a treatment regimen where it is given twice daily for 4 days.  相似文献   

PGF2αeCG及hCG 3种生殖激素联用对母犬发情、卵泡发育及胚胎着床的影响   总被引：3，自引：0，他引：3  

谭建华  安铁洙  刘珠果  万忠海  周虚 《中国兽医学报》2002,22(4):394-396

给休情期性成熟母犬肌注 PGF2α(0 .5 m g/ kg) ,2 4 h后肌注 e CG(5 0 IU/ kg) ,再经 12 0 h后肌注 h CG(2 0 U/ kg) ,诱导其发情。将诱导发情的母犬与公犬交配 ,于交配后 16 d,摘取卵巢、子宫 ,观察其卵泡发育和胚胎着床情况。结果表明 ,药物处理可明显促使母犬提前结束休情期而进入发情期 (9/ 10 ) ,促进卵巢内卵泡发育 (P<0 .0 1) ,并使更多的胚胎着床 ,每只母犬子宫胚胎着床点为 (17.4± 2 .2 )个 ,提示 3种生殖激素联合应用可提高母犬繁殖率  相似文献   

鸡促性腺激素释放激素对卵泡膜分泌雌二醇的影响     

赵畅  汪琳仙 《中国农业大学学报》1993,(2):87-92

本实验选用性成熟的京白种蛋鸡,从同一产蛋顺序中取其发育不同阶段的各级卵泡(F_1～F_4),分离膜层,消化为单个的膜细胞,进行短期细胞培养,着重观察鸡促性腺激素释放激素(GnRH)对培养过程中膜细胞雌二醇分泌的影响.用放射免疫法测定细胞培养液中雌二醇的含量,得到以下结果:①未加外源激素处理的对照组细胞,随着卵泡从小到大的发育成熟过程,膜细胞雌二醇的分泌量逐渐降低;②适当剂量的鸡 GnRH-Ⅱ对各级卵泡膜细胞雌二醇的分泌均有促进作用,其中 F_4,F_3,F_2比 F_1更敏感;③加前体物(孕酮或雄烯二酮)之后,再加鸡 GnRH-Ⅱ比单加前体物或单加鸡 GnRH-Ⅱ,膜细胞雌二醇的分泌量增加更明显.实验结果提示,在体外细胞培养的条件下,GnRH-Ⅱ对膜细胞雌二醇的分泌不仅有促进作用,还可能促进雌二醇的合成。  相似文献   

牛腔前卵泡的简易机械分离方法   总被引：9，自引：0，他引：9  

高志花  周虚  高庆华 《中国草食动物》2003,23(4):3-5

采用两种机械法分离牛腔前卵泡。方法一 (M -1) ,皮肤移植刀切割、剪碎、过滤镜下直接捡卵法。方法二 (M -2 ) ,皮质片剪碎、离心、悬浮、过滤镜下捡卵法。M -1平均每卵巢采卵数为 46 9个± 18 2个 ,M -2为 6 8 4个± 12 5个 ;处理时间M -1为 2 5 4min± 6 7min ,M -2为 46 3min± 16 8min ,两种方法采集腔前卵泡数量及处理时间均存在显著差异。平均每小时采集卵泡数分别为 99 5和 87 2个 ,M -1多于M -2。两种方法获得腔前卵泡大小分布规律也有明显差异 ,M -1采集腔前卵泡直径为 6 0～ 15 0 μm ,绝大部分是次级卵泡 ,而M -2获取腔前卵泡则偏小。总的看来 ,M -1处理时间短 ,回收效率高 ,方法简便 ,且分离卵泡直径较大 ,适于体外培养 ,是机械分离牛腔前卵泡的一种理想方法。  相似文献   

Influence of organic phosphorus on reproductive performance and metabolic profiles of anoestrous Farafra ewes in subtropics at the end of breeding season          下载免费PDF全文

W Senosy  AY Kassab  HA Hamdon  AA Mohammed 《Reproduction in domestic animals》2018,53(4):904-913

The effect of organic phosphorus on metabolic, haematological and hormonal status, restoration of ovarian functions and conception rate in anoestrous Farafra ewes in subtropics were evaluated. Anoestrous Farafra ewes (n  = 24; 34.72 ± 0.52 kg body weight) were allocated into two equal groups: control and phosphorus groups. The ewes of phosphorus group were treated with sodium 4‐dimethylamino‐2‐methyl‐phenyl‐phosphonate as an organic bound phosphorous twice a week for successive 3 weeks. Ovarian follicle development and corpora lutea were checked three times a week till occurrence of oestrus using ultrasonography while pregnancy was confirmed at 30 days post‐service. Plasma metabolites, reproductive hormones, thyroid hormones and minerals were detected at weeks ?2, ?1, 0 (mating day) and + 4 weeks post‐oestrus. Phosphorus group had significantly (p  < .05) short interval to oestrous resumption if compared to control ewes (2.1 ± 0.8 weeks vs . 4.6 ± 1.1 weeks). In addition, phosphorous supplementation significantly (p  < .05) increased the number of antral follicles (developed and their sizes in addition to sizes of corpora lutea (8.72 ± 0.3 mm vs . 7.46 ± 0.9 mm) as well. Number of services per conception (2.6 vs . 1.4; p  < .01) was higher in control group than that of phosphorus group. Pregnancy rate (80 vs . 50%) was significantly (p  < .01) higher in phosphorus group when compared to control. White blood cells in treated ewes (10.8 ± 0.44; p  < .05) and monocytes (2.93 ± 0.13; p  < .01) were higher than that of control group (white blood cells; 9.53 ± 0.50 and monocytes; 2.24 ± 0.14). Metabolic parameters did not differ between phosphorus and control groups during different times of treatment. It could be concluded that phosphorous administration to anoestrous Farafra ewes in subtropics could improve reproductive performance and restore ovarian activity at the end of spring and early summer.  相似文献   

不同培养方法及培养密度对绵羊腔前卵泡发育的影响     

常迪  董晓晨  郭勇  刘云海  齐晓龙  盛熙辉  倪和民 《中国农学通报》2017,33(8):130-134

通过对绵羊腔前卵泡培养方法的研究,能够探索出适宜绵羊腔前卵泡生长发育的培养条件,以及为进一步揭示绵羊卵泡生长发育的规律奠定一些基础。试验一通过使用微滴法和琼脂培养基法培养腔前卵泡,观察其对绵羊腔前卵泡生长的影响;试验二通过绵羊腔前卵泡单独培养与群体培养,探讨不同培养密度对绵羊腔前卵泡生长的影响。试验结果显示,使用微滴法与琼脂培养基法培养的绵羊腔前卵泡,在培养3天时,卵泡直径增加值和存活率差异均不显著(P0.05);但培养6天时,琼脂培养基法培养的腔前卵泡的直径增加值和存活率均极显著高于微滴法培养的腔前卵泡(P0.01);在使用琼脂培养基培养的条件下,对绵羊腔前卵泡进行单独培养和群体培养,在培养的6天时间内二者卵泡直径增加值和存活率差异均不显著(P0.05)。试验结果表明,琼脂培养基法是较为适宜的绵羊腔前卵泡体外培养方法;培养密度对绵羊腔前卵泡体外培养并无显著影响。  相似文献   

NPFFR1基因对鹅卵泡颗粒细胞激素分泌和细胞凋亡的影响研究   总被引：1，自引：1，他引：0  

张克山  高广亮  李琴  赵献芝  李静  王启贵 《畜牧兽医学报》2021,52(10):2822-2831

神经相关肽受体（RFamide-related peptide receptor,NPFFR1）是促性腺激素抑制激素的主要亲和受体,它在调控动物繁殖方面起着重要作用。为了解NPFFR1对鹅卵巢卵泡发育的作用,本研究以42周龄健康产蛋四川白鹅为试验材料（n=9）,利用RT-qPCR法检测NPFFR1基因在等级前和等级卵泡颗粒细胞中的mRNA表达规律;在颗粒细胞中过表达NPFFR1基因,酶联免疫吸附法检测颗粒细胞上清液（n=9）中雌二醇（estradiol,E2）、孕酮（progesterone,P4）和抗缪勒管激素（anti-Mullerian hormone,AMH）的浓度变化,剩余贴壁细胞作一步法TUNEL检测细胞凋亡情况;转录组测序方法筛选大黄卵泡（8~10 mm）颗粒细胞过表达NPFFR1前后表达差异显著基因,并对差异表达基因进行功能聚类分析。结果显示,除F1等级外,其余等级卵泡颗粒细胞NPFFR1表达量均极显著高于等级前卵泡（P<0.01）;过表达NPFFR1后,等级颗粒细胞上清液中的E2和等级前颗粒细胞上清液AMH的含量显著（P<0.05）降低,但孕酮P4含量变化不显著（P>0.05）;转录组测序共筛选到267个差异表达基因（119个下调,148个上调）,这些基因主要富集在生物节律过程、繁殖进程等生物学过程中;同时,与对照组相比,差异基因AMH显著下调表达（P<0.05）,Clock（clock circadian regulator）、FOS（proto-oncogene,AP-1 trans-cription factor subunit）、Per（period circadian regulator）和ANTXR2（cell adhesion molecule 2）分别极显著（P<0.01）或显著（P<0.05）上调表达。上述试验结果提示,NPFFR1可从激素、细胞凋亡和生物节律等多个环节影响卵泡颗粒细胞,参与调控卵泡的时序等级发育。  相似文献   

獭兔毛密度变化及毛囊分化研究     

傅祥超  张凯  文斌  刘宁  刘汉中 《中国养兔杂志》2012,(1):4-7

为了了解獭兔在不同月龄毛囊和毛密度的变化,选择1～7月龄的獭兔为研究对象,通过活体取样,Sacpic法染色,对不同阶段切片进行观察统计,结果表明:毛密度在1～3月龄处于平台期,无显著变化(P0.05);4～7月龄毛密度处于一个波形变动期,其中通过切片观察5～7月龄为全休止期;4～6月龄为换毛期。本研究摸清了毛密度的变化趋势、变化阶段、各阶段毛囊群特点,为獭兔取皮时间的确定及提高毛密度的研究提供理论支撑。  相似文献   

猪卵巢卵母细胞的体外成熟和体外受精   总被引：8，自引：0，他引：8  

邓满齐  谢成侠 《畜牧兽医学报》1992,23(1):1-7

将从屠宰母猪卵巢采得的卵母细胞-卵丘细胞复合体(Oocyte-cumulus Cell Complex.OCC)在含PMSG的M199培养40～44小时,卵丘细胞大部分扩散(86.4％)。48.1％(142/295)的卵母细胞排出第一极体(PBI)。将体外成熟的卵母细胞与体外获能精子授精后30～70小时,80.5％(103/128)的卵母细胞受精并可在体外发育到2～8细胞甚至桑椹胚。本文还对裸卵母细胞的体外成熟和体外受精进行了研究,对体外受精卵的早期发育作了观察。实验结果表明:PMSG对诱导卵丘细胞扩散及卵母细胞的全面成熟有重要作用,在OCC中的卵母细胞成熟率高于裸卵母细胞体外授精后8～10小时将受精卵放入改良KRB液培养可使卵裂比例明显提高。  相似文献