食品科学 ›› 2020, Vol. 41 ›› Issue (9): 98-104.doi: 10.7506/spkx1002-6630-20190426-343

• 营养卫生 • 上一篇    下一篇

葡萄籽原花青素对顺铂诱导小鼠睾丸支持细胞TM4细胞凋亡的影响

杜月梅,韩何丹,郭卓雨,高丽萍   

  1. (1.北京联合大学生物化学工程学院,北京 100023;2.生物活性物质与功能食品北京市重点实验室,北京 100191)
  • 出版日期:2020-05-15 发布日期:2020-05-15
  • 基金资助:
    北京市自然科学基金项目(7163211);生物活性物质与功能食品北京市重点实验室开放课题(LDSP201801)

Effect of Grape Seed Proanthocyanidin Extract on Cisplatin-Induced Apoptosis of Mouse Testicular Sertoli TM4 Cells

DU Yuemei, HAN Hedan, GUO Zhuoyu, GAO Liping   

  1. (1. College of Biochemical Engineering, Beijing Union University, Beijing 100023, China; 2. Beijing Municipal Key Laboratory of Biologically Active Substances and Functional Food, Beijing 100191, China)
  • Online:2020-05-15 Published:2020-05-15

摘要: 目的:研究葡萄籽原花青素(grape seed proanthocyanidin extract,GSPE)对顺铂(cisdichlorodiamineplatinum (II),cDDP)诱导小鼠睾丸支持细胞(TM4)细胞凋亡、细胞凋亡形态及凋亡相关基因蛋白表达的影响。方法:体外培养正常TM4细胞,将实验细胞分为空白对照组、cDDP组(0.014 mg/mL)、GSPE(0.005 mg/mL)+cDDP组(0.014 mg/mL)、GSPE组(0.005 mg/mL)。流式细胞检测法测定GSPE对cDDP诱导TM4细胞凋亡率的影响,Hoechst染液染色法测定TM4细胞凋亡的形态学变化,Western blot蛋白免疫印迹法测定凋亡相关基因蛋白表达情况。结果:cDDP可显著增加TM4细胞凋亡,显著降低TM4细胞Bcl-2表达量以及处于酶原状态Pro-caspase3表达量,而明显增加TM4细胞Bax表达量,同时增加处于激活状态的Caspase-3表达量。GSPE可明显降低cDDP诱导的TM4细胞凋亡,降低Bax以及Caspase-3表达量,增加Bcl-2和Pro-caspase3表达量。结论:细胞凋亡在cDDP诱导TM4细胞毒性过程中起重要作用。Bcl-2、Bax、Caspase-3等蛋白参与cDDP诱导TM4细胞凋亡,GSPE通过抑制Bax及Caspase-3的蛋白表达及增强Bcl-2和Pro-caspase3的表达来阻抑cDDP诱导TM4细胞的凋亡,从而减轻cDDP诱导TM4细胞损伤。

关键词: 葡萄籽原花青素, 顺铂, 小鼠睾丸支持细胞, 细胞凋亡

Abstract: Objective: To study the effect of grape seed procyanidin extract (GSPE) combined with cisplatin (cisdichlorodiamineplatinum (II), cDDP) on cell apoptosis and apoptosis-related protein expression in mouse testicular sertoli TM4 cells. Methods: Normal TM4 cells were cultured in vitro and divided into control, cDDP (0.014 mg/mL), GSPE (0.005 mg/mL) + cDDP (0.014 mg/mL) and GSPE (0.005 mg/mL) group. Flow cytometry was used to determine the effect of GSPE on cDDP-induced cell apoptosis rate, Hoechst staining was used to determine the morphological change of TM4 cells, and Western blot was used to determine the protein expression levels of apoptosis-related genes. Results: cDDP significantly increased the apoptosis of TM4 cells, significantly decreased the expression of the anti-apoptosis proteins Bcl-2 and pro-caspase 3, significantly increased the expression of the pro-apoptosis protein Bax, and increased the expression of activated caspase-3. GSPE could significantly reduce cDDP-induced apoptosis, reduce the expression of Bax and caspase-3, and increase the expression of Bcl-2 and pro-caspase3. Conclusion: Apoptosis plays an important role in TM4 cytotoxicity induced by cDDP. Bcl-2, Bax, caspase-3 and other proteins are involved in the cDDP-induced apoptosis of TM4 cells, which can be inhibited by GSPE through inhibiting the protein expression of Bax and caspase-3 and enhancing the expression of Bcl-2 and pro-caspase 3, thereby reducing cDDP-induced damage to TM4 cells.

Key words: grape seed proanthocyanidin extract, cisplatin, mouse testicular sertoli TM4 cells, cell apoptosis

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