The Mechanism of Midazolam on Mouse Leydig Tumor Cell Apoptosis

• Main Authors: Man-ChiHuang, 黃滿旗
• Other Authors: Bu-Miin Huang
• Format: Others
• Language: en_US
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/54942736076042484756

碩士 === 國立成功大學 === 細胞生物與解剖學研究所 === 101 === Midazolam is widely used as sedative anesthetic induction agent, and it is a derivative from benzodiazepine drug. In previous study, we have found that midazolam could induce steroidogenesis in MA-10 mouse Leydig cells via PKA and PKC pathways along with the expression of PBR and StAR proteins1. In addition, midazolam at higher dosages induced rounding-up, membrane blebbing, and then cell death phenomenon in MA-10 cells. In fact, we have found that midazolam could stimulate caspase and MAPK, but inhibit Akt, pathways to induce apoptosis in MA-10 cells. In the present study, we would like to further investigate the detail mechanism activated by midazolam in MA-10 cells, and will focus on the apopototic pathway-, autophage pathway-, ER strees pathway-, and PKC pathway-related protein expressions. The results showed that midazolam induced the expression of Bax, cytochrome C and Light Chain 3 II (LC3II) proteins, but decreased the expression of Bid, phosphor-Akt and phosphor-mTOR proteins. These data illustrated that midazolam could activate intrinsic apopototic pathway plus autophagy in MA-10 cells. Moreover, midazolam treatment induced the cleavage of caspase-7,-12 and the phosphorylations of PKC, but decreased the expression of p53. These results demonstrated that midazolam might activate ER stress and PKC pathways to induce MA-10 cell death. We also hypothesized that midazolam could induce apoptosis via the mitochondrial pathway in MA-10 cell. We observated that the expressions of intrinsic apoptotic pathway proteins, other autophagy-related proteins, ER stress pathway-related proteins; activating transcription factor 6 (ATF6), X box-binding protein 1 (XBP1), eukaryotic translation initiator factor 2α (eIF2α), and different isoforms of PKC proteins were changed by treatment of midazolam. In conclusion, midazolam could activate apoptotic pathway, ER stress pathway, PKC pathway with autophage involvement to induce MA-10 cell apoptosis.