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[Effects of triptolide on the activity of TM3 Leydig cells and AMPK/Akt/mTOR pathway].

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.