Hydrogen sulfide regulates vascular endoplasmic reticulum stress in apolipoprotein E knockout mice
Hydrogen sulfide regulates vascular endoplasmic reticulum stress in apolipoprotein E knockout mice
复制标题
硫化氢调节载脂蛋白E基因敲除小鼠血管内质网应激
DOI：
10.3760/cma.j.issn.0366-6999.2011.21.005
复制
发表时间：
2011-11-05
影响因子：
6.1
通讯作者：
Jin Hong-fang
中科院分区：
文献类型：
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作者：
Chen Zhi-fang;Zhao Bin;Jin Hong-fang
研究背景动脉粥样硬化是一种重要的心血管疾病，在发达国家已成为日益严重的健康问题。然而，可能的潜在机制并不完全清楚。2009年，本课题组首次发现硫化氢(H_2S)作为一种新型的胃递质具有重要的抗动脉粥样硬化作用。为探讨硫化氢对载脂蛋白E基因敲除小鼠(apoE(-/-))内质网应激(ERS)的调节作用，采用西式饲料喂养C57BL/6小鼠和apoE(-/-)纯合子小鼠。对照组：C57BL/6小鼠腹腔注射生理盐水(5ml/(kg·d))。ApoE(-/-)组为apoE(-/-)+NaHS组，生理盐水剂量与apoE(-/-)组相同；apoE(-/-)+NaHS组为硫化氢供体(NaHS，56mU/kg/d)；10周后处死小鼠，检测血脂。用HE和油红0染色观察主动脉根部的动脉粥样硬化病变。用光镜和电子显微镜观察主动脉的超微结构。免疫组织化学方法检测大鼠主动脉组织葡萄糖调节蛋白78(GRP78)、半胱氨酸天冬氨酸氨基转移酶-12(caspase-12)、铜锌超氧化物歧化酶(铜/锌超氧化物歧化酶)和锰超氧化物歧化酶(MnSOD)蛋白的表达。结果与对照组相比，apoE(-/-)小鼠血浆总胆固醇(TC)、甘油三酯(TG)和低密度脂蛋白(LDL)水平升高，高密度脂蛋白(HDL)水平降低，主动脉斑块增大，主动脉组织超微结构破坏，GRP78和caspase-12蛋白表达增加。与apoE(-/-)小鼠相比，硫化氢供体处理的apoE(-/-)小鼠血浆低密度脂蛋白水平降低，斑块坏死减轻，主动脉超微结构紊乱减轻。硫化氢供体处理可诱导主动脉病变GRP78表达，但抑制caspase-12表达。然而，与apoE(-/-)小鼠相比，PPG治疗的apoE(-/-)小鼠表现为斑块扩大，主动脉组织超微结构紊乱更严重，主动脉病变中GRP78染色减少。ApoE(-/-)+PPG大鼠与apoE(-/-)-小鼠的血脂和caspase-12染色无显著差异。结论硫化氢对高脂饲料喂养的apoE(-/-)小鼠的主动脉ERS具有调节作用，可减轻动脉粥样硬化病变。中华医学杂志2011；124(21)：3460-3467
Background Atherosclerosis is an important cardiovascular disease, becoming a major and increasing health problem in developed countries. However, the possible underlying mechanisms were not completely clear. In 2009, our research group first discovered that hydrogen sulfide (H2S) as a novel gastrotransmitter played an important anti-atherosclerotic role. The study was designed to examine the regulatory effect of hydrogen sulfide (H2S) on endoplasmic reticulum stress (ERS) in apolipoprotein E knockout (apoE(-/-)) mice fed a Western type diet.Methods C57BL/6 mice and homozygous apoE(-/-) mice were fed a Western type diet. C57BL/6 mice were injected intraperitoneally with normal saline (5 ml/kg per day) as control group. The apoE(-/-) mice were treated with the same dose of normal saline as the apoE(-/-) group, injected intraperitoneally with sodium hydrosulfide (NaHS, an H2S donor, 56 mu mol/kg per day) as the apoE(-/-)+NaHS group and injected intraperitoneally with DL-propargylglycine (PPG, a cystathionine-gamma-lyase inhibitor, 50 mg/kg, per day) as the apoE(-/-) +PPG group. After 10 weeks, the mice were sacrificed and the plasma lipids were detected. Sections of aortic root from these animals were examined for atherosclerotic lesions by HE and oil red 0 staining. The aortic ultrastructure and microstructure were analyzed with the help of light and electronic microscope. Glucose-regulated protein 78 (GRP78), caspase-12, copper-andzinc-containing superoxide dismutase (Cu/ZnSOD) and Mn-containing superoxide dismutase (MnSOD) protein expression in aortic tissues were detected with immunohistochemistry. The level of intracellular reactive oxygen species (ROS) were measured by using a commercial assay kit.Results Compared with control mice, apoE(-/-) mice showed increased plasma levels of total cholesterol (TC), triglyceride (TG) and low density lipoprotein (LDL), decreased high density lipoprotein (HDL), increased aortic plaque size, destroyed ultra-structure of aortic tissue, and increased expression of GRP78 and caspase-12 proteins. Compared with apoE(-/-) mice, H2S donor-treated apoE(-/-) mice showed a decreased plasma LDL level, lessened plaque necrosis and attenuated aortic ultra-structural disorder. H2S donor-treatment induced GRP78 expression but suppressed caspase-12 expression in aortic lesions. However, compared with apoE(-/-) mice, PPG treated apoE(-/-) mice showed enlarged plaque size, more severe ultrastructural disorder of the aortic tissue and reduced GRP78 staining in aortic lesions. The plasma lipids and the staining of caspase-12 in apoE(-/-) + PPG rats did not significantly differ from those in the apoE(-/-)-mice. Consistently, H2S induced SOD expression, accompanied by a reduced level of ROS.Conclusion H2S plays a regulatory role in aortic ERS and reduces atherosclerotic lesions in apoE(-/-) mice fed with a Western type diet. Chin Med J 2011;124(21):3460-3467