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[Effects of triptolide on the activity of TM3 Leydig cells and AMPK/Akt/mTOR pathway]
1Center of Reproduction and Genetics, Department of Gynecology and Obstetrics, Peking University First Hospital, Beijing 100034, China.
Objective:
To investigate the effects of triptolide (TP) on the activity of TM3 Leydig cells and the AMPK/Akt/mTOR pathway.
Methods:
We treated TM3 Leydig cells with TP at 50, 100 and 200 nmol/L, respectively, and with 0.1% DMSO as the control, and cultured them in a 37℃ thermostat container for 24 hours. Then we measured the damage to the cell membrane and apoptosis of the cells using lactate dehydrogenase (LDH) activity assay and the apoptosis assay kit, and detected the changes in the AMPK/Akt/mTOR pathway-related proteins by Western blot.
Results:
The LDH activity was significantly increased in the 50, 100 and 200 nmol/L TP groups in a dose-dependent manner compared with that in the DMSO control (［163.4±33.6］%, ［346.8±148.8］% and ［422.8±113.9］% vs ［157.5±20.3］%, P < 0.01), and so was the apoptosis of the cells (［189.9±73.5］%, ［284±103.5］% and ［419.2±155.7］% vs ［6.27±1.41］%, P < 0.01). The TP-treated cells showed a significantly decreased p-AMPK/AMPK ratio in comparison with the control (P < 0.01), but an increased p-Akt/Akt ratio (P < 0.05) and an elevated level of p-mTOR phosphorylation (P < 0.05).
Conclusions:
Triptolide can decrease the activity and induce the apoptosis of TM3 Leydig cells, and meanwhile inhibit the AMPK and activate the Akt/mTOR pathway.
Insights
Triptolide (TP) reduces TM3 Leydig cell activity and induces apoptosis. It inhibits the AMPK pathway while activating the Akt/mTOR pathway, impacting cell function.
Area of Science:
- Cell Biology
- Molecular Biology
- Endocrinology
Background:
- TM3 Leydig cells are crucial for testosterone production.
- The AMPK/Akt/mTOR pathway plays a vital role in cellular metabolism and survival.
- Triptolide is a compound with known biological activities.
Purpose of the Study:
- To examine the impact of triptolide on TM3 Leydig cell viability.
- To investigate the effects of triptolide on the AMPK/Akt/mTOR signaling pathway.
Main Methods:
- TM3 Leydig cells were treated with varying concentrations of triptolide (50-200 nmol/L).
- Cell membrane damage was assessed via lactate dehydrogenase (LDH) activity.
- Apoptosis and protein expression (AMPK, Akt, mTOR) were analyzed using Western blot.
Main Results:
- Triptolide significantly increased LDH activity and induced apoptosis in a dose-dependent manner.
- A decrease in the p-AMPK/AMPK ratio was observed.
- An increase in p-Akt/Akt ratio and p-mTOR levels indicated pathway activation.
Conclusions:
- Triptolide impairs TM3 Leydig cell function and promotes cell death.
- Triptolide disrupts the AMPK pathway while activating the Akt/mTOR signaling cascade.
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