来曲唑对睾丸间质细胞系TM3 增殖、凋亡能力的影响及作用机制
| 作者: |
1王澍弘,
1李行,
2李晶,
1李美材,
1李晓霞,
1龙腾博,
1王顺德
1 重庆三峡中心医院男性科，重庆 404000 2 重庆三峡中心医院中心实验室，重庆 404000 |
| 通讯: |
王顺德
Email: 635784196@qq.com |
| DOI: | 10.3978/j.issn.2095-6959.2018.10.002 |
| 基金: | 重庆市卫生和计划生育委员会项目 (2016MSXM114)。 |
摘要
目的：研究来曲唑(letrozole，LE)对睾丸间质细胞系TM3增殖、凋亡能力和丝裂原活化蛋白激酶(mitogen activated protein kinase，MAPK)信号通路的影响。方法：采用不同浓度的LE(1，10， 100，1 000 μmol/L)处理小鼠睾丸间质细胞TM3，细胞计数(Cell Counting Kit-8，CCK-8)法检测细胞增殖。采用LE(100 μmol/L)和MAPK信号通路抑制剂APS-2-79(50 nmol/L)分别单独或联合作用于小鼠睾丸间质细胞TM3 48 h，CCK-8法检测细胞增殖，流式细胞术检测细胞凋亡，Western印迹检测MAPK信号通路关键蛋白总细胞外调节蛋白激酶(extracellular regulated protein kinases，ERK)和磷酸化细胞外调节蛋白激酶(phosphorylation of extracellular regulated protein kinases，p-ERK)的表达。结果：LE浓度在1.0~100.0 μmol/L时，从作用的第24小时开始，能够显著促进小鼠睾丸间质细胞TM3(P<0.05)，具有时间和剂量依赖效应。小鼠睾丸间质细胞TM3培养48 h，LE组细胞的OD450值显著高于对照组和LE+APS-2-79组(P<0.05)。而APS-2-79组的细胞的OD450值显著低于对照组(P<0.05)。LE组细胞凋亡率显著低于对照组和LE+APS-2-79组(P<0.05)。而APS-2-79组的细胞凋亡率显著高于对照组(P<0.05)。LE组ERK和p-ERK蛋白表达以及p-ERK与ERK比值显著高于对照组和LE+APS-2-79组(P<0.05)；而APS-2-79组的ERK和p-ERK蛋白表达以及p-ERK与ERK比值显著低于对照组(P<0.05)。结论：LE能够促进小鼠睾丸间质细胞TM3的增殖，抑制其凋亡，其作用机制可能与MAPK信号通路相关。
关键词：
来曲唑；增殖；凋亡；丝裂原活化蛋白激酶信号通路
Effect of letrozole on the proliferation and apoptosis of TM3 in mesenchymal cell line of testis and its mechanism
CorrespondingAuthor: WANG Shunde Email: 635784196@qq.com
DOI: 10.3978/j.issn.2095-6959.2018.10.002
Foundation: This work was supported by the Chongqing Health and Family Planning Commission Projects, China (2016MSXM114).
Abstract
Objective: To study the effect of letrozole (LE) on the proliferation, apoptosis and mitogen activated protein kinase (MAPK) signaling pathway of interstitial cell line TM3. Methods: Testicular mesenchymal cells (TM3) of mice were treated with LE at different concentrations (1, 10, 100, 1 000 μmol/L), and cell proliferation was detected by Cell Counting Kit-8 (CCK-8). LE (100 μmol/L) and MAPK signal pathway inhibitor APS-2-79 (50 nmol/L) were alone or combined respectively in mouse leydig cells TM3 48 h, CCK-8 method to detect cell proliferation and flow cytometry to detect cell apoptosis, Western blot to test key protein ERK and p-ERK. Results: When LE concentration was 1.0–100.0 μmol/L, TM3 of mouse testicular stromal cells (P<0.05) could be significantly promoted from the 24th h of the effect, with time and dose dependent effect. Mouse testicular mesenchymal cells were cultured for 48 h, and the OD450 value of the cells in the LE group was significantly higher than that in the control group and the LE+ APS-2-79 group (P<0.05). The OD450 value of cells in APS-2-79 group was significantly lower than that of control group (P<0.05). The apoptosis rate of cells in LE group was significantly lower than that in control group and LE+APS-2-79 group (P<0.05). The apoptosis rate of cells in APS-2-79 group was significantly higher than that of control group (P<0.05). The expression levels of ERK and p-ERK, the ratio of p-ERK and ERK in the LE group were significantly higher than those in the control group and the LE+APS-2-79 group (P<0.05), while the expression levels of ERK and p-ERK, the ratio of p-ERK and ERK in the APS-2-79 group were significantly lower than those in the control group. Conclusion: LE can promote the proliferation of mouse testicular interstitial cells TM3 and inhibit its apoptosis, and its mechanism may be related to MAPK signaling pathway.
Keywords：
letrozole; proliferation; apoptosis; mitogen activated protein kinase signaling pathway