Ethanol Withdrawal Provokes Opening of the Mitochondrial Membrane Permeability Transition Pore in an Estrogen-Preventable Manner
Ethanol Withdrawal Provokes Opening of the Mitochondrial Membrane Permeability Transition Pore in an Estrogen-Preventable Manner
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DOI：
10.1124/jpet.108.146829
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发表时间：
2009-03-01
影响因子：
3.5
通讯作者：
Simpkins, James W.
中科院分区：
文献类型：
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作者：
Jung, Marianna E.;Wilson, Andrew M.;Simpkins, James W.
我们已经报道，主要的内源性雌激素，17 β-雌二醇（E2），防止氧化损伤乙醇戒断（EW）在培养的海马细胞系（HT 22）。在这里，我们调查是否亲氧化性质的EW介导开放的线粒体膜通透性转换孔（PTP）的方式保护E2。过度的PTP开放引起线粒体膜肿胀（MMS）和膜电位的崩溃（Delta Psi m）。在乙醇暴露（100 mM）24 h结束时或EW 4 h时收集HT 22细胞，以通过监测540 nm处的吸光度下降来评估MMS，并使用流式细胞术评估Δ Psi m。将E2对PTP的保护作用与抗氧化剂丁基化羟基甲苯（BHT）和E2类似物ZYC 26 [（3-羟基-2-金刚烷基（1）-4-甲基-雌甾-1，3，5（10）-17-酮]进行了比较，其抗氧化效力高于E2。为了评估PTP开放的细胞后果，使用钙黄绿素测定评估PTP抑制剂（环孢菌素A）对EW诱导的细胞死亡的影响。主要结论如下：1）EW导致快速MMS和Δ Psim崩溃; 2）环孢菌素A减弱EW诱导的细胞死亡;和3）限制于EW期的E2处理比BHT更显著地保护PTP开放，并且达到与ZYC 26相似的程度。这些研究结果表明，EW引起PTP开放部分但不完全通过促氧化剂的性质，E2抵消EW相关的因素，以防止PTP开放。
We have reported that the major endogenous estrogen, 17 beta-estradiol (E2), protects against oxidative injury during ethanol withdrawal (EW) in a cultured hippocampal cell line (HT22). Here, we investigated whether the pro-oxidant nature of EW mediates opening of the mitochondrial membrane permeability transition pore (PTP) in a manner protected by E2. Excess PTP opening provokes mitochondrial membrane swelling (MMS) and the collapse of membrane potential (Delta Psi m). HT22 cells were collected at the end of ethanol exposure (100 mM) for 24 h or at 4 h of EW to assess MMS by monitoring absorbance decline at 540 nm and to assess Delta Psi m using flow cytometry. Protective effects of E2 on PTP were compared with an antioxidant butylated hydroxytoluene (BHT) and an E2 analog, ZYC26 [( 3-hydroxy-2-adamantyl(1)-4-methyl-estra-1,3,5(10)-17-one], with higher antioxidant potency than E2. To assess cellular consequences of PTP opening, effects of a PTP inhibitor (cyclosporin A) on EW-induced cell death were assessed using the calcein assay. Major findings were that: 1) EW resulted in rapid MMS and Delta Psi m collapse; 2) cyclosporin A attenuated EW-induced cell death; and 3) E2 treatment restricted to the EW phase protected against the PTP opening more prominently than BHT and to a similar degree to ZYC26. These findings suggest that EW provokes PTP opening partly but not entirely through the pro-oxidant nature and that E2 counteracts EW-associated factors to protect against the PTP opening.