Tetrahydrocurcumin ameliorates homocysteinylated cytochrome-c mediated autophagy in hyperhomocysteinemia mice after cerebral ischemia.
Tetrahydrocurcumin ameliorates homocysteinylated cytochrome-c mediated autophagy in hyperhomocysteinemia mice after cerebral ischemia.
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DOI：
10.1007/s12031-011-9695-z
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发表时间：
2012-05
期刊：
影响因子：
--
通讯作者：
Tyagi SC
中科院分区：
文献类型：
--
作者：
Tyagi N;Qipshidze N;Munjal C;Vacek JC;Metreveli N;Givvimani S;Tyagi SC
高水平的同型半胱氨酸（Hcy）被称为高同型半胱氨酸血症（hyperhomocysteine-mia，HHcy），有助于自噬和缺血/再灌注损伤（ischemia/reperfusion injury，I/R）。既往研究表明I/R损伤和高同型半胱氨酸可导致脑血管通透性增加，但相关机制尚不清楚。有趣的是，在HHcy期间，细胞色素c变成同型半胱氨酸化（Hcy-cyto-c）。细胞色素c（cyto-c）传输电子并促进系统中的生物能量学。然而，其在缺血/再灌注损伤中的自噬作用尚不清楚。四氢姜黄素（THC）是一种主要的草药抗氧化剂和抗炎剂。因此，本研究的目的是确定THC是否通过减少高同型半胱氨酸血症病理条件下cyto-c的同型半胱氨酸化来改善缺血/再灌注损伤期间的自噬。为了验证这一假设，我们采用了8-10周龄的雄性胱硫醚-β-合酶杂合子敲除（CBS+/−）小鼠（遗传性高同型半胱氨酸小鼠）。实验组为：CBS+/-、CBS+/-+THC（25 mg/kg，含0.1%DMSO剂量）; CBS（+/-）/I/R和CBS（+/-）/I/R+THC（25 mg/kg，含0.1%DMSO剂量）。缺血30 min，再灌注72 h。腹腔内（I. P.）缺血30天后每天一次，持续3天。用氯化三苯基四氮唑染色法测量梗死面积。通过脑水肿和伊文思蓝外渗测定渗透性。通过Western blot分析脑组织的氧化应激、基质金属蛋白酶-9（MMP-9）、损伤调节的自噬调节剂（DRAM）和微管相关蛋白1轻链3（LC 3）。采用实时定量聚合酶链反应检测S-腺苷-L-同型半胱氨酸水解酶（SAHH）和亚甲基四氢叶酸还原酶（MTHFR）基因mRNA水平。采用免疫共沉淀法测定cyto-c的同型半胱氨酸化。我们发现，与假手术组相比，I/R+THC治疗组的脑水肿和伊文思蓝渗漏减少沿着脑梗死面积减少。与假手术组相比，THC还减少了I/R组的氧化损伤，并通过激活MMP-9部分改善了细胞C的同型半胱氨酸化，从而导致自噬。这项研究表明，饮食THC在脑缺血中具有潜在的治疗作用。
High levels of homocysteine (Hcy) known as hyperhomocysteine-mia (HHcy), contribute to autophagy and ischemia/reperfusion injury (I/R). Previous studies have shown that I/R injury and HHcy cause increased cerebrovascular permeability; however, the associated mechanism remains obscure. Interestingly, during HHcy, cytochome-c becomes homocysteinylated (Hcy-cyto-c). Cytochrome-c (cyto-c) transports electrons and facilitates bioenergetics in the system. However, its role in autophagy during ischemia/reperfusion injury is unclear. Tetrahydrocurcumin (THC) is a major herbal antioxidant and anti-inflammatory agent. Therefore, the objective of this study was to determine whether THC ameliorates autophagy during ischemia/reperfusion injury by reducing homocysteinylation of cyto-c in hyperhomocysteinemia pathological condition. To test this hypothesis we employed 8–10 weeks old male cystathionine-beta-synthase heterozygote knockout (CBS+/−) mice (genetically hyperhomocystemic mice). Experimental group was: CBS+/−, CBS+/−+THC (25mg/kg in 0.1%DMSO dose); CBS (+/−)/I/R and CBS (+/−)/I/R+THC(25mg/kg in 0.1%DMSO dose). Ischemia was performed for 30 min and reperfusion for 72 hours. THC was injected intra-peritoneally (I.P.) once daily for a period of 3 days after 30 of ischemia. The infarct area was measured using 2,3,5-triphenyltetrazolium chloride staining. Permeability was determined by brain edema and Evans Blue extravasation. The brain tissues were analyzed for oxidative stress, matrix metalloproteinase-9 (MMP-9), damage-regulated autophagy modulator (DRAM), and microtubule-associated protein 1 light chain 3 (LC3) by Western blot. The mRNA levels of S-Adenosyl-L-homocysteine hydrolases (SAHH) and Methylenetetrahydrofolate reductase (MTHFR) genes were measured by Quantitative real-time polymerase chain reaction. Co-immunoprecipitation was used to determine the homocysteinylation of cyto-c. We found that brain edema and Evans Blue leakage were reduced in I/R+THC treated groups as compared to sham operated groups along with reduced brain infarct size. THC also decreased oxidativedamage and ameliorated the homocysteinylation of cyto-c in-part by MMP-9 activation which leads to autophagy in I/R groups as compared to sham operated groups. This study suggests a potential therapeutic role of dietary THC in cerebral ischemia.
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影响因子：
4.1
作者：
Lee, JE;Yoon, YJ;Yenari, MA
通讯作者：
Yenari, MA
影响因子：
2.9
作者：
Belayev, L;Busto, R;Ginsberg, MD
通讯作者：
Ginsberg, MD
DOI：
10.1097/01.wcb.0000088762.02615.30
发表时间：
2003-10-01
影响因子：
6.3
作者：
Fukui, S;Fazzina, G;Marmarou, A
通讯作者：
Marmarou, A
影响因子：
5.3
作者：
Lee, SR;Tsuji, K;Lo, EH
通讯作者：
Lo, EH
影响因子：
6.8
作者：
Jakubowski, Hieronim
通讯作者：
Jakubowski, Hieronim