Malondialdehyde and 4-hydroxynonenal adducts are not formed on cardiac ryanodine receptor (RyR2) and sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA2) in diabetes.
Malondialdehyde and 4-hydroxynonenal adducts are not formed on cardiac ryanodine receptor (RyR2) and sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA2) in diabetes.
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DOI：
10.1007/s11010-013-1558-1
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发表时间：
2013-04
影响因子：
4.3
通讯作者：
Bidasee KR
中科院分区：
文献类型：
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作者：
Moore CJ;Shao CH;Nagai R;Kutty S;Singh J;Bidasee KR
最近，我们报道了链脲佐菌素（STZ）诱导的糖尿病大鼠心脏中心脏兰尼碱受体（RyR2）和肌（内）质网Ca2+-ATP酶（SERCA2）上葡萄糖衍生的羰基加合物水平升高。我们还表明这些加合物损害了 RyR2 和 SERCA2 活性，并改变了诱发的 Ca2+ 瞬变。尚不清楚的是，脂质衍生的丙二醛 (MDA) 和 4-羟基-2-壬烯醛 (4-HNE) 是否也会与糖尿病中的 RyR2 和 SERCA2 发生化学反应并损害其活性。本研究使用加合物特异性抗体和共聚焦显微镜进行蛋白质印迹测定，以评估 RyR2 和 SERCA2 上的 MDA、4-HNE、Nε-羧基（甲基）赖氨酸 (CML)、戊糖苷和吡咯啉加合物的水平，并在对照、糖尿病和治疗糖尿病大鼠的肌细胞中诱发细胞内瞬时 Ca2+ 动力学。在对照大鼠或诱发 Ca2+ 瞬变改变的 8 周糖尿病大鼠的 RyR2 和 SERCA2 上未检测到 MDA 和 4-HNE 加合物。然而，CML、戊糖素和吡咯啉加合物升高了 3-5 倍（p<0.05）。用吡哆胺（一种活性羰基物质清除剂，RCS）或氨基胍（一种混合活性氧-活性羰基物质清除剂）治疗糖尿病大鼠可减少 RyR2 和 SERCA2 上的 CML、戊糖苷和吡咯啉加合物，并减弱 SR Ca2+ 循环变化。用超氧化物歧化酶模拟 tempol 治疗糖尿病大鼠对 RyR2 和 SERCA2 上的 MDA 和 4-HNE 加合物以及 SR Ca2+ 循环没有影响。根据这些数据，我们得出结论，在该糖尿病模型中，脂质衍生的 MDA 和 4-HNE 加合物并未在 RyR2 和 SERCA2 上形成，因此不太可能直接导致 SR Ca2+ 失调。
Recently, we reported an elevated level of glucose-derived carbonyl adducts on cardiac ryanodine receptor (RyR2) and sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA2) in hearts of streptozotocin(STZ)-induced diabetic rats. We also showed these adduct impaired RyR2 and SERCA2 activities, and altered evoked Ca2+ transients. What is less clear is if lipid-derived malondialdehyde (MDA) and 4-hydroxy-2-nonenal (4-HNE) also chemically react with and impair RyR2 and SERCA2 activities in diabetes. This study used Western blot assays with adduct-specific antibodies and confocal microscopy to assess levels of MDA, 4-HNE, Nε-carboxy(methyl)lysine (CML), pentosidine and pyrraline adducts on RyR2 and SERCA2 and evoked intracellular transient Ca2+ kinetics in myocytes from control, diabetic and treated-diabetic rats. MDA and 4-HNE adducts were not detected on RyR2 and SERCA2 from control or 8 weeks diabetic rats with altered evoked Ca2+ transients. However, CML, pentosidine, and pyrraline adducts were elevated 3–5 fold (p<0.05). Treating diabetic rats with pyridoxamine (a scavenger of reactive carbonyl species, RCS) or aminoguanidine (a mixed reactive oxygen species-reactive carbonyl species scavenger) reduced CML, pentosidine and pyrraline adducts on RyR2 and SERCA2 and blunted SR Ca2+ cycling changes. Treating diabetic rats with the superoxide dismutase mimetic tempol had no impact on MDA and 4-HNE adducts on RyR2 and SERCA2, and on SR Ca2+ cycling. From these data we conclude that lipid-derived MDA and 4-HNE adducts are not formed on RyR2 and SERCA2 in this model of diabetes, and are therefore unlikely to be directly contributing to the SR Ca2+ dysregulation.