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CdCl2与尼古丁对精原细胞联合作用方式的初步研究
作者:
作者单位:

1.山西医科大学公共卫生学院;2.深圳市疾病预防控制中心深圳市现代毒理学重点实验室

基金项目:

深圳市医疗卫生三名工程项目(SZSM202211010),深圳市医学重点学科建设经费资助项目(SZXK069)


Preliminary studies on the mode of CdCl2 in combination with nicotine on spermatogonial cells
Author:
Affiliation:

1.College of Public Health, Shanxi Medical University,Shenzhen Key Labora⁃ tory of Modern Toxicology,Shenzhen Medical Key Discipline of Health Toxicology;2.Shenzhen Key Labora⁃ tory of Modern Toxicology,Shenzhen Medical Key Discipline of Health Toxicology

Fund Project:

Supported by Sanming Project of Medicine in Shenzhen（SZSM202211010）,Shenzhen Medical Key Discipline Construction Funding Program(SZXK069)

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    摘要:

    目的：探讨CdCl2与联合尼古丁对小鼠精原GC-1细胞周期和凋亡的影响及其联合作用方式。方法：CCK-8法检测CdCl2组、尼古丁组以及联合作用组24h时对GC-1细胞增殖的抑制,流式细胞术检测CdCl2组、尼古丁组以及联合作用组24h时对GC-1细胞周期和凋亡的影响。 结果：随着CdCl2、尼古丁浓度的升高,对GC-1细胞增殖的抑制作用增加,CdCl2对GC-1细胞的24h IC50为5.409μmol/L,尼古丁对GC-1细胞的24h IC50为2814μmol/L。CdCl2和尼古丁联用时,尼古丁浓度为0.175mmol/L时,GC-1细胞的IC50为4.422μmol/L；尼古丁浓度为0.35mmol/L时,GC-1细胞的IC50为4.532μmol/L；尼古丁浓度为0.7mmol/L时,GC-1细胞的IC50为3.309μmol/L,尼古丁浓度为1.4mmol/L时,GC-1细胞的IC50为2.532μmol/L,均显著低于CdCl2单独做用组；CdCl2浓度为2.5μmol/L时联用组IC50浓度低于尼古丁单独作用浓度,CdCl2和尼古丁联合作用时具有协同作用。细胞周期结果显示,CdCl2浓度为2.5 μmol/L和尼古丁浓度为1.4mmol/L时G0/G1期细胞占比下降,与CdCl2组、尼古丁组比较,CdCl2和尼古丁联合作用组G0/G1期细胞占比下降；CdCl2浓度为2.5μmol/L时G2/M期细胞占比上升,与CdCl2组、尼古丁组比较,CdCl2和和尼古丁联合作用时将细胞阻滞在G2/M期。CdCl2和尼古丁联合作用时对细胞周期的影响具有协同作用,且CdCl2对于协同作用的影响更强。细胞凋亡结果显示,CdCl2浓度为2.5μmol/L时,细胞凋亡比例增加,与CdCl2组、尼古丁组单独作用时比较,CdCl2和尼古丁联合作用组细胞凋亡比例显著增加。CdCl2和尼古丁联合作用时对细胞凋亡的影响具有协同作用,且CdCl2对于协同作用的影响更强。结论：CdCl2和尼古丁联合时具有协同作用,使细胞毒性作用增强,细胞周期发生阻滞,促进细胞的凋亡,并且CdCl2对协同作用的影响更强。

    Abstract:

    Objective：To investigate the combined effect of CdCl2 and nicotine on the cell cycle and apoptosis of mouse spermatogonium(GC-1).Methods：CCK-8 assay was used to detect the inhibition of GC-1 cell proliferation in the CdCl2 groups, the nicotine groups, and the combined groups at 24 hours. Flow cytometry was used to detect the effects of the CdCl2 groups, the nicotine groups, and the combined groups at 24 hours on the cell cycle and apoptosis of GC-1 cells. Results：The inhibitory effect on the proliferation of GC-1 cells was increased with the rise of CdCl2 and nicotine concentrations , the IC50 values of CdCl2 and nicotine alone was 5.409μmol/L, 2814μmol/L. When CdCl2 and nicotine were used in combination, the IC50 value of GC-1 cells was 4.422 μmol/L for 0.175 mmol/L nicotine, 4.532 μmol/L for 0.35 mmol/L nicotine, 3.309 μmol/L for 0.7 mmol/L nicotine, and 2.532 μmol/L for 1.4 mmol/L nicotine ,all of which were lower than that of the groups in which CdCl2 was used alone. The combination of CdCl2 and nicotine had a synergistic effect. At a CdCl2 concentration of 2.5 μmol/L, the IC50 concentration of the combination group was lower than that of nicotine alone. The results showed that percentage of G0/G1 phase cells was decreased in the 2.5 μmol/L CdCl2 group and 1.4 mmol/L nicotine group; compared with the CdCl2 groups and the nicotine groups, percentage of G0/G1 phase cells was decreased in the CdCl2 and nicotine combination groups; percentage of G2/M phase cells was increased in the 2.5 μmol/L CdCl2 group, and compared with the CdCl2 groups and nicotine groups, the cells were blocked in G2/M phase in the combined groups. Combined effects of CdCl2 and nicotine on the cell cycle had a synergistic effect , and the effect of CdCl2 on the synergistic effect was stronger. Proportion of cell apoptosis was increased in 2.5μmol/L CdCl2 group. Compared with the CdCl2 groups and the nicotine groups, percentage of cell apoptosis in the CdCl2 and nicotine combined groups were increased significantly, and the effect of CdCl2 was stronger for the synergistic effect. Conclusion：Combination of CdCl2 and nicotine had a synergistic effect on mouse spermatogonium(GC-1) which could lead to enhanced cytotoxicity, G2/M cell cycle arrests, and cell apoptosis. And CdCl2 was the main effect factor of synergistic effect.

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  • 收稿日期:2024-05-09
  • 最后修改日期:2024-06-26
  • 录用日期:2024-07-11
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