本文關(guān)鍵詞： 膿毒癥 心肌損傷 線粒體自噬 微管相關(guān)蛋白輕鏈(LC) pink parkin　出處：《細(xì)胞與分子免疫學(xué)雜志》2016年02期 　論文類型：期刊論文

【摘要】：目的探討膿毒癥心肌損傷時(shí)心肌細(xì)胞及線粒體自噬水平的變化。方法雄性C57BL/J小鼠隨機(jī)分為空白對(duì)照組(NC)、LPS處理6、12、24、36 h組。LPS處理組小鼠腹腔注射10 mg/kg LPS建立膿毒癥模型,空白對(duì)照組注射等量生理鹽水。分別于以上時(shí)間點(diǎn)處死小鼠,收集血液及心臟組織,提取心肌組織胞質(zhì)蛋白、線粒體及線粒體蛋白,另取心肌組織進(jìn)行冰凍切片;應(yīng)用ELISA試劑盒測定血清心肌肌鈣蛋白I(c Tn I)含量變化;JC-1染色結(jié)合熒光酶標(biāo)儀檢測LPS刺激后不同時(shí)間點(diǎn)線粒體膜電位變化;Western blot法檢測自噬相關(guān)蛋白微管相關(guān)蛋白1輕鏈3(LC3)、PTEN誘導(dǎo)激酶1(pink1)、E3泛素連接酶帕金森病蛋白(parkin)水平;免疫熒光組織化學(xué)染色檢測心肌組織LC3、pink1/parkin蛋白的定位及其在不同處理組的表達(dá)差異。結(jié)果與對(duì)照組相比,LPS誘導(dǎo)的膿毒癥小鼠血清c Tn I在6 h即開始明顯升高;LPS處理組線粒體膜電位下降,在LPS 12 h組最低;細(xì)胞自噬相關(guān)蛋白LC3-Ⅱ/LC3-Ⅰ在腹腔注射LPS 12 h明顯升高,然后逐漸下降;pink1/parkin在腹腔注射LPS 6 h明顯升高,然后逐漸下降。結(jié)論膿毒癥心肌損傷時(shí)心肌細(xì)胞及線粒體自噬水平升高,自噬應(yīng)激可能更早發(fā)于心肌線粒體。
[Abstract]:Objective to investigate the changes of cardiac myocyte and mitochondrial autophagy during myocardial sepsis. Methods male C57BL / J mice were randomly divided into two groups: control group (n = 36) treated with LPS-treated with LPS-61212O / 24 h. LPS-treated mice were injected intraperitoneally with 10 mg/kg LPS to establish septic model. Mice were killed at the above time points, blood and heart tissues were collected, myocardial cytoplasmic protein, mitochondria and mitochondrial protein were extracted, and myocardial tissue was taken for frozen section. Detection of changes of serum cardiac troponin I c Tn I by ELISA kit and detection of mitochondrial membrane potential at different time points after LPS stimulation by using JC-1 staining and fluorescence enzyme labeling. Western blot method for the detection of autophagy associated protein microtubule-associated protein 1. The level of PTEN-induced kinase 1 Pink1PK E3 ubiquitin ligase Parkinsonase (Parkinin) was detected in light chain L3 LC3 and PTEN. Immunofluorescence histochemical staining was used to detect the localization of LC3pink1 / parkin protein and its expression in different treatment groups. Results compared with the control group, the serum c TN I of LPS induced sepsis mice began to increase significantly at 6 h after lipopolysaccharide (LPS) treatment. The mitochondrial membrane potential decreased in the group, In LPS 12h group, LC3- 鈪，

本文編號(hào)：1502449