梓醇对顺铂诱导大鼠卵巢颗粒细胞损伤的保护作用
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摘要
目的探讨梓醇对顺铂诱导大鼠卵巢颗粒细胞损伤的保护作用及作用机制。方法收集大鼠卵巢颗粒细胞,经原代培养后设为对照组、模型组(顺铂50 nmol/L)和梓醇50、100、200μmol/L组,各组细胞按相应方式进行处理。采用MTT法测定细胞活力;流式细胞术Annexin V/PI双染法考察细胞凋亡情况;免疫组化法观察细胞增殖能力,WB法定量细胞凋亡蛋白的分子水平。结果顺铂可显著抑制卵巢颗粒细胞活力、增殖,诱导细胞凋亡;在给予梓醇进行干预后,大鼠卵巢颗粒细胞活力增强、增殖上调,细胞凋亡减弱;经顺铂进行刺激损伤,卵巢颗粒细胞中caspase-3、caspase-9的活化蛋白表达量显著上升,而不同浓度的梓醇可显著下调细胞中cleaved caspase-3、cleaved caspase-9的蛋白表达量。结论梓醇可以改善大鼠细胞活力、提高细胞增殖和抗凋亡能力,对顺铂诱导的卵巢颗粒细胞损伤产生明显的保护作用。
Objective: To investigate protective effects of catalpolon on apoptosis of rat ovarian granulosa cells induced by cisplatin, and explore its mechanisms. Methods Rat ovarian granulosa cells were cultured and divided into control group, model group(cisplatin 50 nmol/L), and catalpol(50, 100, and 200 μmol/L) groups. Then cells in each group were treated in the corresponding ways. Cell viability was determined by MTT method. Apoptosis was detected by double staining flow cytometry Annexin V/PI. Western blotting method was used to detect expressions of apoptosis related protein. Results Cisplatin could significantly decrease the viability of rat ovarian granulosa cells and induce apoptosis. After treated by catalpol, the activity of ovarian granulosa cells was increased, proliferation was up-regulated, and apoptosis was decreased. After treated by cisplatin, the expressions of the active form of cleaved caspase-3 and cleaved caspase-9 protein were significantly increased. Catalpol could significantly decrease the expressions of cleaved caspase-3 and cleaved caspase-9 in cells. Conclusion The addition of catalpol to rat ovarian granulosa cells can significantly change the anti-apoptotic and anti-oxidative ability, and play a protective role in the apoptosis induced by cisplatin. Catalpol can improve cell viability, enhance cell proliferation and anti apoptosis ability, and protect apoptosis of ovarian granulosa cells in rats induced by cisplatin.
Objective: To investigate protective effects of catalpolon on apoptosis of rat ovarian granulosa cells induced by cisplatin, and explore its mechanisms. Methods Rat ovarian granulosa cells were cultured and divided into control group, model group(cisplatin 50 nmol/L), and catalpol(50, 100, and 200 μmol/L) groups. Then cells in each group were treated in the corresponding ways. Cell viability was determined by MTT method. Apoptosis was detected by double staining flow cytometry Annexin V/PI. Western blotting method was used to detect expressions of apoptosis related protein. Results Cisplatin could significantly decrease the viability of rat ovarian granulosa cells and induce apoptosis. After treated by catalpol, the activity of ovarian granulosa cells was increased, proliferation was up-regulated, and apoptosis was decreased. After treated by cisplatin, the expressions of the active form of cleaved caspase-3 and cleaved caspase-9 protein were significantly increased. Catalpol could significantly decrease the expressions of cleaved caspase-3 and cleaved caspase-9 in cells. Conclusion The addition of catalpol to rat ovarian granulosa cells can significantly change the anti-apoptotic and anti-oxidative ability, and play a protective role in the apoptosis induced by cisplatin. Catalpol can improve cell viability, enhance cell proliferation and anti apoptosis ability, and protect apoptosis of ovarian granulosa cells in rats induced by cisplatin.
引文
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[11]Wang Z,Liu Q,Zhang R.Catalpol ameliorates beta amyloidinduced degeneration of cholinergic neurons by elevating brain-derived neurotrophic factors[J].Neuroscience,2009,163(4):1363-1372.
[12]Zhang X,Jin C,Li Y,et al.Catalpol improves cholinergic function and reduces inflammatory cytokines in the senescent mice induced by D-galactose[J].Food Chem Toxicol,2013,58(7):50-55.
[13]Choi H J,Jang H J,Chung T W,et al.Catalpol suppresses advanced glycation end-products-induced inflammatory responses through inhibition of reactive oxygen species in human monocytic THP-1 cells[J].Fitoterapia,2013,86(1):19-28.
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[16]Gao N,Tian J X,Shang Y H,et al.Catalpol suppresses proliferation and facilitates apoptosis of OVCAR-3ovarian cancer cells through upregulating micro RNA-200and downregulating MMP-2 expression[J].Int J Mol Sci,2014,15(11):19394-19405.
[17]马丽灵,阮祥燕.卵巢早衰病因及治疗研究进展[J].医学综述,2008,14(23):3557-3560.
[18]王静欢,邹利,万东,等.梓醇多效性相关信号通路研究进展[J].中国药理学通报,2015,31(9):1189-1194.
[19]董炤,陈长勋.梓醇药理作用的研究进展[J].中成药,2013,35(5):1047-1051.
[20]刘艳茹.梓醇预处理对脑缺血再灌注损伤的神经保护作用研究[D].郑州:郑州大学,2014.
[21]刘晨阳,白宽,于顾然.梓醇对纤维状Aβ1-42诱导的b End.3细胞凋亡及过度自噬的影响[J].中国实验方剂学杂志,2017,23(20):108-113.
[22]Scholzen T,Gerdes J.The Ki-67 protein:from the known and the unknown[J].J Cell Physiol,2000,182(3):311-322.
[23]Meral O,Ozgur E,Kismali G,et al.GSM-like radiofrequency exposure induces apoptosis via caspase-dependent pathway in infant rabbits[J].Bratisl Lek Listy,2016,117(11):672-676.
[24]金娟,马林沁,邹国良,等.参芪益心方对慢性心力衰竭大鼠心肌组织Caspase-3及Caspase-9蛋白表达的影响[J].世界中西医结合杂志,2014,9(2):151-155.
[2]Dzafic E,Stimpfel M,Virant-Klun I.Plasticity of granulosa cells:on the crossroad of stemness and transdifferentiation potential[J].J Assist Reprod Genet,2013,30(10):1255-1261.
[3]艾浩,牛建昭,薛晓鸥,等.顺铂致小鼠卵巢功能早衰肝肾阴虚证机制研究[J].北京中医药大学学报,2006,29(6):401-403.
[4]Sanchez A M,Giorgione V,ViganòP,et al.Treatment with anticancer agents induces dysregulation of specific Wnt signaling pathways in human ovarian luteinized granulosa cells in vitro[J].Toxicol Sci,2013,136(1):183-192.
[5]Sun L,Li D,Song K,et al.Exosomes derived from human umbilical cord mesenchymal stem cells protect against cisplatin-induced ovarian granulosa cell stress and apoptosis in vitro[J].Sci Rep,2017,7(1):2552.
[6]石彩歌.中西医结合治疗卵巢早衰的临床观察[J].中医临床研究,2011,03(15):34-35.
[7]中国药典[S].一部.2010:115-116.
[8]Li D Q,Duan Y L,Bao Y M,et al.Neuroprotection of catalpol in transient global ischemia in gerbils[J].Neurosci Res,2004,50(2):169-177.
[9]Li D Q,Bao Y M,Zhao J J,et al.Neuroprotective properties of catalpol in transient global cerebral ischemia in gerbils:dose-response,therapeutic time-window and long-term efficacy[J].Brain Res,2004,1029(2):179-185.
[10]石桂芳,王贺双,毛玉荣,等.梓醇对鱼藤酮所致小鼠脑线粒体损伤的保护作用[J].现代生物医学进展,2012,12(29):5661-5664.
[11]Wang Z,Liu Q,Zhang R.Catalpol ameliorates beta amyloidinduced degeneration of cholinergic neurons by elevating brain-derived neurotrophic factors[J].Neuroscience,2009,163(4):1363-1372.
[12]Zhang X,Jin C,Li Y,et al.Catalpol improves cholinergic function and reduces inflammatory cytokines in the senescent mice induced by D-galactose[J].Food Chem Toxicol,2013,58(7):50-55.
[13]Choi H J,Jang H J,Chung T W,et al.Catalpol suppresses advanced glycation end-products-induced inflammatory responses through inhibition of reactive oxygen species in human monocytic THP-1 cells[J].Fitoterapia,2013,86(1):19-28.
[14]Bi J,Jiang B,Zorn A,et al.Catalpol inhibits LPS plus IFN-γ-induced inflammatory response in astrocytes primary cultures[J].Toxicol in Vitro,2013,27(2):543-550.
[15]Pungitore C R,León L G,García C,et al.Novel antiproliferative analogs of the Taq DNA polymerase inhibitor catalpol[J].Bioorg Med Chem Lett,2007,17(5):1332-1335.
[16]Gao N,Tian J X,Shang Y H,et al.Catalpol suppresses proliferation and facilitates apoptosis of OVCAR-3ovarian cancer cells through upregulating micro RNA-200and downregulating MMP-2 expression[J].Int J Mol Sci,2014,15(11):19394-19405.
[17]马丽灵,阮祥燕.卵巢早衰病因及治疗研究进展[J].医学综述,2008,14(23):3557-3560.
[18]王静欢,邹利,万东,等.梓醇多效性相关信号通路研究进展[J].中国药理学通报,2015,31(9):1189-1194.
[19]董炤,陈长勋.梓醇药理作用的研究进展[J].中成药,2013,35(5):1047-1051.
[20]刘艳茹.梓醇预处理对脑缺血再灌注损伤的神经保护作用研究[D].郑州:郑州大学,2014.
[21]刘晨阳,白宽,于顾然.梓醇对纤维状Aβ1-42诱导的b End.3细胞凋亡及过度自噬的影响[J].中国实验方剂学杂志,2017,23(20):108-113.
[22]Scholzen T,Gerdes J.The Ki-67 protein:from the known and the unknown[J].J Cell Physiol,2000,182(3):311-322.
[23]Meral O,Ozgur E,Kismali G,et al.GSM-like radiofrequency exposure induces apoptosis via caspase-dependent pathway in infant rabbits[J].Bratisl Lek Listy,2016,117(11):672-676.
[24]金娟,马林沁,邹国良,等.参芪益心方对慢性心力衰竭大鼠心肌组织Caspase-3及Caspase-9蛋白表达的影响[J].世界中西医结合杂志,2014,9(2):151-155.