姜黄素对水牛卵巢颗粒细胞体外培养的影响
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摘要
本试验研究了添加姜黄素对水牛卵巢颗粒细胞体外培养过程中增殖、凋亡自噬及间隙连接等生物学过程的影响。设置不同浓度(1,5,10,25,50,100μg/m L)的姜黄素,对颗粒细胞的凋亡、增殖、自噬和间隙连接等基因与蛋白的表达进行定性与定量检测,发现高浓度姜黄素会导致细胞凋亡标记Caspase-3及自噬基因Beclin-1的基因表达量上升,间隙连接蛋白Cx43基因表达量下降,细胞增殖标记PCNA表达量无显著变化。研究结果为姜黄素在调节卵巢颗粒细胞功能提供了一定的理论依据。
In this research,we studied the effects of curcumin on the proliferation,apoptosis autophagy and connexin of buffalo ovarian granule cells in vitro. By setting different concentrations(1,5,10,25,50,100 μg/m L),the gene and protein expressions including apoptosis,proliferation,autophagy and connexin of granulosa cells were identified through qualitative and quantitative analyses. The results showed that the expression of apoptotic marker Caspase-3 and autophagy gene Beclin-1 increased and the expression of the connexin-Cx43 gene decreased with the higher concentration of curcumin. The expression of PCNA showed no significant alteration. The present findings indicated that high concentration of curcumin(100 μg/m L) induced the apoptosis and cell death of buffalo ovarian granule cells in vitro,and also affected the growth of cells in vitro.
In this research,we studied the effects of curcumin on the proliferation,apoptosis autophagy and connexin of buffalo ovarian granule cells in vitro. By setting different concentrations(1,5,10,25,50,100 μg/m L),the gene and protein expressions including apoptosis,proliferation,autophagy and connexin of granulosa cells were identified through qualitative and quantitative analyses. The results showed that the expression of apoptotic marker Caspase-3 and autophagy gene Beclin-1 increased and the expression of the connexin-Cx43 gene decreased with the higher concentration of curcumin. The expression of PCNA showed no significant alteration. The present findings indicated that high concentration of curcumin(100 μg/m L) induced the apoptosis and cell death of buffalo ovarian granule cells in vitro,and also affected the growth of cells in vitro.
引文
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[14] Li M Q,Luo A L,Zhao P W,et al. Nanos2 is a molecular marker of inchoate buffalo spermatogonia[J]. Animal Reproduction Science,2017:44-51.
[15] Durlinger A L,Kramer P,Karels B,et al. Apoptotic and proliferative changes during induced atresia of pre-ovulatory follicles in the rat[J]. Human Reproduction,2000,15:2504-2511.
[16] Tiu V K,Bryzhina T M,Sukhina V S,et al. Effect of NF-kappaB activation inhibitor curcumin on the oogenesis and follicular cell death in immune ovarian failure in mice[J]. Fiziologicheskii Zhurnal Imeni I. M. Sechenova,2010,56:96-101.
[17] Kádasi A,MaruniakováN,tochmaíováA,et al. Direct effect of curcumin on porcine ovarian cell functions[J]. Animal Reproduction Science,2017:77-83.
[18] Shang J H,Yang C Y,Zheng H Y,et al. Effects of curcumin on buffalo embryonic development in vitro[J]. Buffalo Bulletin,2014,32:456-459.
[2] Ak T,Gulcin I. Antioxidant and radical scavenging properties of curcumin[J]. Chemico-Biological Interactions,2008,174:27-37.
[3] Tiwari-Pandey R,Ram S M. Modulation of ovarian structure and abdominal obesity in curcumin-and flutamide-treated aging FSH-R haploinsufficient mice[J]. Reproductive Sciences, 2009,16:539.
[4] Terlikowska K M,Witkowska A M,Zujko M E,et al. Potential application of curcumin and its analogues in the treatment strategy of patients with primary epithelial ovarian cancer[J]. International Journal of Molecular Sciences,2014,15:21703-21722.
[5] Shehzad A,Wahid F,Lee Y S. Curcumin in cancer chemoprevention:molecular targets,pharmacokinetics,bioavailability,and clinical trials[J]. Archiv Der Pharmazie,2010,343:489.
[6] Watson J L,Greenshields A,Hill R,et al. Curcumin-induced apoptosis in ovarian carcinoma cells is p53-independent and involves p38 mitogen-activated protein kinase activation and downregulation of Bcl-2 and survivin expression and Akt signaling[J]. Molecular Carcinogenesis,2010,49:13.
[7] Vallianou N G,Evangelopoulos A,Schizas N,et al. Potential anticancer properties and mechanisms of action of curcumin[J]. Anticancer Research,2015,35:645.
[8] Seo J A,Kim B,Dhanasekaran D N,et al. Curcumin induces apoptosis by inhibiting sarco/endoplasmic reticulum ca(2+)ATPase activity in ovarian cancer cells[J]. Cancer Letters,2016,371:30.
[9] Lin L,Zakeri Z F. Apoptosis in development[J]. Nature,2000,136:107.
[10]李玲,徐小洁,叶棋浓.细胞自噬与肿瘤[J].中国生物化学与分子生物学报,2013,29:995-1001.
[11] Levine E M,Becker Y,Boone C W,et al. Contact inhibition,macromolecular synthesis,and polyribosomes in cultured human diploid fibroblasts[J]. Proc Natl Acad Sci U S A,1965,53:350-356.
[12] Mehta P P,Bertram J S,Loewenstein W R. Growth inhibition of transformed cells correlates with their junctional communication with normal cells[J]. Cell,1986,44:187.
[13] Zheng H Y,Yang C Y,Shang J H,et al. The impact of linolenic acid on in vitro development of buffalo embryos[J]. Buffalo Bulletin,2014,32:432-435.
[14] Li M Q,Luo A L,Zhao P W,et al. Nanos2 is a molecular marker of inchoate buffalo spermatogonia[J]. Animal Reproduction Science,2017:44-51.
[15] Durlinger A L,Kramer P,Karels B,et al. Apoptotic and proliferative changes during induced atresia of pre-ovulatory follicles in the rat[J]. Human Reproduction,2000,15:2504-2511.
[16] Tiu V K,Bryzhina T M,Sukhina V S,et al. Effect of NF-kappaB activation inhibitor curcumin on the oogenesis and follicular cell death in immune ovarian failure in mice[J]. Fiziologicheskii Zhurnal Imeni I. M. Sechenova,2010,56:96-101.
[17] Kádasi A,MaruniakováN,tochmaíováA,et al. Direct effect of curcumin on porcine ovarian cell functions[J]. Animal Reproduction Science,2017:77-83.
[18] Shang J H,Yang C Y,Zheng H Y,et al. Effects of curcumin on buffalo embryonic development in vitro[J]. Buffalo Bulletin,2014,32:456-459.