红景天甙干预可改善慢性间断性缺氧模型小鼠的肺损伤
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摘要
背景:阻塞性睡眠呼吸暂停低通气综合征和慢性阻塞性肺疾病是呼吸系统常见疾病,二者并存发生率高,临床上称之为"重叠综合征",慢性间断性缺氧是重叠综合征的基本病变过程。目的:探讨红景天甙对慢性间断性缺氧小鼠细胞凋亡和氧化应激肺损伤的作用机制。方法:实验项目经西南医科大学动物实验伦理委员会批准。取野生型C57BL/6雄性小鼠随机分为正常对照组、红景天甙组、慢性间断性缺氧组、慢性间断性缺氧+红景天甙组,后两组模拟慢性间断性缺氧模型:即将小鼠放入缺氧舱,调节初始氧浓度8%-10%维持1min后迅速转为常氧(19%-21%)持续2min为一个完整循环,每天循环8 h,连续30 d。缺氧当天起红景天甙组、慢性间断性缺氧+红景天甙组小鼠给予红景天甙50 mg/(kg·d)灌胃处理连续30 d。光镜下观察肺组织病理变化;化学荧光法定量、定性检测小鼠肺组织氧自由基活性;Western-blot法检测肺组织凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶3(caspase-3)、Bax、Bcl-2水平及Bax/Bcl-2比值。结果与结论:(1)小鼠发生慢性间断性缺氧时激活肺氧化应激途径并启动细胞凋亡,表现为与正常对照组相比,慢性间断性缺氧组病理损伤显著,肺动脉管壁增厚,肺泡大小不均,活性氧活性升高,Bax/Bcl-2比值、caspase-3蛋白水平升高(P<0.05);而给予红景天甙处理可有效改善慢性间断性缺氧造成的小鼠肺损伤,与慢性间断性缺氧组相比,慢性间断性缺氧+红景天甙组肺病理损伤减轻,活性氧活性降低、凋亡蛋白Bax/Bcl-2比值、caspase-3水平下降(P <0.05);正常对照组与红景天甙组各项指标无明显差异(P> 0.05);(2)结果说明,红景天甙可以通过抑制氧化应激反应和细胞凋亡途径改善小鼠慢性间断性缺氧肺损伤。
BACKGROUND: Obstructive sleep apnea hypopnea syndrome and chronic obstructive pulmonary disease are common diseases in respiratory system. The incidence of coexistence is high, which is called "overlap syndrome" in clinical practice. Chronic intermittent hypoxia is the basic pathological process of overlap syndrome.OBJECTIVE: To investigate the mechanism of salidroside affecting the apoptosis and oxidative stress in lung injury caused by chronic intermittent hypoxia in mice.METHODS: The study was approved by the Ethics Committee of Laboratory Animal Center of Southwest Medical University. Wild-type C57 BL/6 male mice were randomly divided into normal control group, salidroside group, chronic intermittent hypoxia group and chronic intermittent hypoxia+salidroside group. The mice in the latter two groups were placed in an anoxic chamber. The complete cycle of hypoxia was adjusted to the initial oxygen concentration of 8% to 10% for 1 minute, and then rapidly converted to normoxia(19%-21%) for 2 minutes.The daily cycle of hypoxia was 8 hours for 30 days. On the day of hypoxia, mice in the salidrosidel and salidrosidel groups were given salidroside(50 mg/(kg·d) for 30 consecutive days. The pathological changes of lung tissue were observed under a light microscope.Quantitative and qualitative detection of oxygen free radical activity in mouse lung tissue was performed by chemical fluorescence method.The apoptosis related protein caspase-3, Bax, Bcl-2 levels, and Bax/Bcl-2 ratio were detected by western blot assay.RESULTS AND CONCLUSION:(1) The occurrence of chronic intermittent hypoxia activated the lung oxidative stress pathway and initiated cell apoptosis. The lung pathological damage in the chronic intermittent hypoxia group was more serious than that in the normal control group.Its pulmonary artery wall thickened in the normal control group, the alveolar size was uneven, the oxygen free radical activity was increased,the ratio of Bax/Bcl-2 and the protein level of caspase-3 were significantly increased(P < 0.05). However, the degree of lung pathological damage in the chronic intermittent hypoxia+salidroside group was significantly reduced and the above indicators in the chronic intermittent hypoxia+salidroside group were significantly lower than those in the chronic intermittent hypoxia group(P < 0.05). There was no significant difference in each indicator between normal control and salidroside groups(P > 0.05).(2) Therefore, salidroside may improve chronic intermittent hypoxic lung injury in mice by inhibiting oxidative stress response and apoptotic pathways.
BACKGROUND: Obstructive sleep apnea hypopnea syndrome and chronic obstructive pulmonary disease are common diseases in respiratory system. The incidence of coexistence is high, which is called "overlap syndrome" in clinical practice. Chronic intermittent hypoxia is the basic pathological process of overlap syndrome.OBJECTIVE: To investigate the mechanism of salidroside affecting the apoptosis and oxidative stress in lung injury caused by chronic intermittent hypoxia in mice.METHODS: The study was approved by the Ethics Committee of Laboratory Animal Center of Southwest Medical University. Wild-type C57 BL/6 male mice were randomly divided into normal control group, salidroside group, chronic intermittent hypoxia group and chronic intermittent hypoxia+salidroside group. The mice in the latter two groups were placed in an anoxic chamber. The complete cycle of hypoxia was adjusted to the initial oxygen concentration of 8% to 10% for 1 minute, and then rapidly converted to normoxia(19%-21%) for 2 minutes.The daily cycle of hypoxia was 8 hours for 30 days. On the day of hypoxia, mice in the salidrosidel and salidrosidel groups were given salidroside(50 mg/(kg·d) for 30 consecutive days. The pathological changes of lung tissue were observed under a light microscope.Quantitative and qualitative detection of oxygen free radical activity in mouse lung tissue was performed by chemical fluorescence method.The apoptosis related protein caspase-3, Bax, Bcl-2 levels, and Bax/Bcl-2 ratio were detected by western blot assay.RESULTS AND CONCLUSION:(1) The occurrence of chronic intermittent hypoxia activated the lung oxidative stress pathway and initiated cell apoptosis. The lung pathological damage in the chronic intermittent hypoxia group was more serious than that in the normal control group.Its pulmonary artery wall thickened in the normal control group, the alveolar size was uneven, the oxygen free radical activity was increased,the ratio of Bax/Bcl-2 and the protein level of caspase-3 were significantly increased(P < 0.05). However, the degree of lung pathological damage in the chronic intermittent hypoxia+salidroside group was significantly reduced and the above indicators in the chronic intermittent hypoxia+salidroside group were significantly lower than those in the chronic intermittent hypoxia group(P < 0.05). There was no significant difference in each indicator between normal control and salidroside groups(P > 0.05).(2) Therefore, salidroside may improve chronic intermittent hypoxic lung injury in mice by inhibiting oxidative stress response and apoptotic pathways.
引文
[1]Zhang S,ChenX,YangY,et al.Neuroprotection against cobalt chloride-induced cell apoptosis of primary cultured cortical neurons by salidroside.Mol Cell Biochem 2011;354(1-2):161-170.
[2]马生龙,边惠萍,朱芳一,等.红景天甙对低氧条件下H9C2心肌细胞凋亡及相关蛋白Caspase-3表达的影响[J].高原医学杂志,2016,26(1):1-4.
[3]史佩,杨立霞,王楠.红景天苷对低氧条件下大鼠心肌细胞凋亡及相关蛋白表达的影响[J].中华中医药杂志,2018,33(9):4195-4199.
[4]黄薇,荣黎,陈耀凯,欧阳净,等.红景天甙对人胃癌细胞增殖及凋亡影响的研究[J].中国免疫学杂志,2019,35(6):676-679+685.
[5]黄献欢,黄维,高兴华.红景天甙抑制大鼠肺纤维化机制研究[J].临床和实验医学杂志,2018,17(4):343-345.
[6]姚立军,刘媛.红景天甙对慢性阻塞性肺疾病小鼠保护作用机制研究[J].中国药学杂志,2017,52(17):44-47.
[7]Ding W,Cai Y,Wang W,et al.Adiponectin protects the kidney against chronic intermittent hypoxia-induced injury through inhibiting endoplasmic reticulum stress.Sleep Breath 2016;20(3):1069-1074
[8]赵岩,杨丹,于珊珊.黄芪甲苷对慢性心力衰竭大鼠心肌细胞凋亡及P-Cx43表达水平的影响[J].临床和实验医学杂志,2018,17(20):2143-2147.
[9]陈小雨,高飞,钱永常.姜黄素在肿瘤治疗中的应用机制[J].临床合理用药杂志,2019,12(9):175-176.
[10]吴万征,李朝晖,梁球.西藏红景天对小鼠辐射损伤的保护作用及其抗高原反应与低温环境的作用[J].中药材,2005,28(2):128-130.
[11]Wu XL,Zeng WZ,Wang PL,et al.Effect of compound rhodiolasachalinensis A Bor on CCl4-induced liver fibrosis in rats and its probable molecular mechanisms.World journal of gastroenterology.2003;9(7):1559-1562.
[12]欧阳金生,李玉苹,蔡畅,等.红景天苷对人肺动脉内皮细胞线粒体ROS及细胞分泌功能的影响[J].中华中医药学刊,2013,31(4):798-800.
[13]张金平,汪丽云,王永梅,等.红景天苷对缺氧诱导乳鼠海马神经元凋亡的抑制作用及机制[J].山东医药,2018,58(37):16-19.
[14]贾真.红景天苷对大鼠急性心肌缺血的保护作用[J].中医研究,2015,28(8):57-59
[15]Xu J,Qian J,Xie X,et al.High Density Lipoprotein Protects Mesenchymal Stem Cells from Oxidative Stress-Induced Apoptosis via Activation of the PI3K/Akt Pathway and Suppression of Reactive Oxygen Species.Int J Mol Sci.2012;13(12):17104-17120.
[16]ChereshP,KimSJ,TulasiramS,etal.Oxidative stress and pulmonary fibrosis.Biochimica Et Biophysica Acta.2013;1832(7):1028-1040.
[17]Santos JH,HunakovaL,ChenY,et al.Cell Sorting Experiments Link Persistent Mitochondrial DNA Damage with Loss of Mitochondrial Membrane Potential and Apoptotic Cell Death.JBiol Chem.2003;278(3):1728-1734.
[18]Sinha K,DasJ,PalPB,etal.Oxidative stress:The mitochondriadependent and mitochondria-independent pathways of apoptosis.ArchToxicol.2013;87(7):1157-1180.
[19]Pressly JD,Mustafa SM,AbidiA,etal.Selective cannabinoid 2 receptor stimulation reduces tubular epithelial cell damage following renal ischemia-reperfusion injury.JPharmacolExpTher.2017(2).
[20]赵诗洁.红景天甙对鸡胚肺血管内皮细胞?肺血管平滑肌细胞及心肌细胞中HIF-1α表达的影响[D].武汉:华中农业大学,2014.
[2]马生龙,边惠萍,朱芳一,等.红景天甙对低氧条件下H9C2心肌细胞凋亡及相关蛋白Caspase-3表达的影响[J].高原医学杂志,2016,26(1):1-4.
[3]史佩,杨立霞,王楠.红景天苷对低氧条件下大鼠心肌细胞凋亡及相关蛋白表达的影响[J].中华中医药杂志,2018,33(9):4195-4199.
[4]黄薇,荣黎,陈耀凯,欧阳净,等.红景天甙对人胃癌细胞增殖及凋亡影响的研究[J].中国免疫学杂志,2019,35(6):676-679+685.
[5]黄献欢,黄维,高兴华.红景天甙抑制大鼠肺纤维化机制研究[J].临床和实验医学杂志,2018,17(4):343-345.
[6]姚立军,刘媛.红景天甙对慢性阻塞性肺疾病小鼠保护作用机制研究[J].中国药学杂志,2017,52(17):44-47.
[7]Ding W,Cai Y,Wang W,et al.Adiponectin protects the kidney against chronic intermittent hypoxia-induced injury through inhibiting endoplasmic reticulum stress.Sleep Breath 2016;20(3):1069-1074
[8]赵岩,杨丹,于珊珊.黄芪甲苷对慢性心力衰竭大鼠心肌细胞凋亡及P-Cx43表达水平的影响[J].临床和实验医学杂志,2018,17(20):2143-2147.
[9]陈小雨,高飞,钱永常.姜黄素在肿瘤治疗中的应用机制[J].临床合理用药杂志,2019,12(9):175-176.
[10]吴万征,李朝晖,梁球.西藏红景天对小鼠辐射损伤的保护作用及其抗高原反应与低温环境的作用[J].中药材,2005,28(2):128-130.
[11]Wu XL,Zeng WZ,Wang PL,et al.Effect of compound rhodiolasachalinensis A Bor on CCl4-induced liver fibrosis in rats and its probable molecular mechanisms.World journal of gastroenterology.2003;9(7):1559-1562.
[12]欧阳金生,李玉苹,蔡畅,等.红景天苷对人肺动脉内皮细胞线粒体ROS及细胞分泌功能的影响[J].中华中医药学刊,2013,31(4):798-800.
[13]张金平,汪丽云,王永梅,等.红景天苷对缺氧诱导乳鼠海马神经元凋亡的抑制作用及机制[J].山东医药,2018,58(37):16-19.
[14]贾真.红景天苷对大鼠急性心肌缺血的保护作用[J].中医研究,2015,28(8):57-59
[15]Xu J,Qian J,Xie X,et al.High Density Lipoprotein Protects Mesenchymal Stem Cells from Oxidative Stress-Induced Apoptosis via Activation of the PI3K/Akt Pathway and Suppression of Reactive Oxygen Species.Int J Mol Sci.2012;13(12):17104-17120.
[16]ChereshP,KimSJ,TulasiramS,etal.Oxidative stress and pulmonary fibrosis.Biochimica Et Biophysica Acta.2013;1832(7):1028-1040.
[17]Santos JH,HunakovaL,ChenY,et al.Cell Sorting Experiments Link Persistent Mitochondrial DNA Damage with Loss of Mitochondrial Membrane Potential and Apoptotic Cell Death.JBiol Chem.2003;278(3):1728-1734.
[18]Sinha K,DasJ,PalPB,etal.Oxidative stress:The mitochondriadependent and mitochondria-independent pathways of apoptosis.ArchToxicol.2013;87(7):1157-1180.
[19]Pressly JD,Mustafa SM,AbidiA,etal.Selective cannabinoid 2 receptor stimulation reduces tubular epithelial cell damage following renal ischemia-reperfusion injury.JPharmacolExpTher.2017(2).
[20]赵诗洁.红景天甙对鸡胚肺血管内皮细胞?肺血管平滑肌细胞及心肌细胞中HIF-1α表达的影响[D].武汉:华中农业大学,2014.