| 张利敏,刘雨佳,陈云秋,张苗苗,张 甦,赵 帅,王新琢,张文妍,郝丽英,焦光宇.钙蛋白酶抑制剂Calpeptin在脓毒症膈肌功能障碍中的作用研究[J].,2022,(5):808-813 |
| 钙蛋白酶抑制剂Calpeptin在脓毒症膈肌功能障碍中的作用研究 |
| The Effect of Calpeptin Inhibitor on Diaphragmatic Dysfunction under Sepsis in Rats |
| 投稿时间：2021-09-23 修订日期：2021-10-18 |
| DOI：10.13241/j.cnki.pmb.2022.05.002 |
| 中文关键词: 脓毒症 膈肌 钙蛋白酶抑制剂 作用机制 保护 |
| 英文关键词: Sepsis Diaphragm Calpeptin Mechanism Protection |
| 基金项目:国家自然科学基金项目（21677030）；辽宁省重点研发攻关项目 (2017225009)；沈阳市科技计划项目(202054021) |
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| 中文摘要: |
| 摘要 目的：探究钙蛋白酶抑制剂Calpeptin在脓毒症膈肌功能障碍中的保护作用及其分子机制。方法：通过腹腔内注射8 mg/kg LPS的方法构建大鼠脓毒症模型，将雄性SD大鼠分为3组（n=6/组）：正常对照组（Con组）、脓毒症组（Sepsis组）和钙蛋白酶抑制剂预处理组（Calpeptin组）。处死大鼠，快速分离大鼠膈肌组织，采用苏木精-伊红(HE)染色法检测膈肌组织病理学改变，通过实时荧光定量PCR法（qRT-PCR）分别检测膈肌组织中钙蛋白酶μ-Calpain、凋亡相关基因天冬氨酸特异性半胱氨酸蛋白酶-3（Caspase-3）、自噬相关蛋白Beclin-1、肿瘤坏死因子-α（TNF-α）和白介素-6（IL-6）mRNA表达水平。结果：与正常对照组相比，LPS处理24 h的脓毒症组大鼠膈肌组织HE染色未见明显膈肌萎缩改变，但膈肌收缩力下降，这与我们以往的研究结果一致。qRT-PCR法检测到脓毒症组大鼠膈肌组织中上述基因mRNA表达量明显增加（P<0.05），而Calpeptin预处理后，上述基因mRNA表达水平显著下降（P<0.05）。结论：脓毒症时膈肌发生功能障碍，钙蛋白酶抑制剂Calpeptin可显著减轻LPS诱导的炎症、凋亡、自噬的激活。 |
| 英文摘要: |
| ABSTRACT Objective: This investigation was under taken to explore probable mechanisms and signal pathways involved in diaphragm dysfunction induced by Sepsis. Methods: The rat sepsis model was constructed by injecting 8 mg/kg LPS into the abdominal cavity. Male SD rats were divided into 3 groups (n=6/group): comparing control group (Con group), LPS group (Sepsis group) and calcium Protease inhibitor pretreatment group(Calpeptin group). Each group of rats was sacrificed, and then their diaphragm tissues were quickly separated. The pathological changes of the diaphragm tissue were observed by using the hematoxylin-eosin (HE) staining method. Additionally, the mRNA expression levels of μ-Calpain, caspase-3, Beclin-1, TNF-α and IL-6 in diaphragm tissue were detected by real-time fluorescence quantitative PCR(qRT-PCR). Results: Under the light microscope, we could see that the diaphragm cross-section in the Con group was regular, with full and orderly muscle fibers, closely spaced muscle fibers, and nuclei distributed along the edge of the lens. In the diaphragm muscle cells of the Con group, there were intact capillary endothelium and no interstitial hyperplasia. Compared with the Con group, HE staining of the sepsis group rats with LPS for 24 h showed no obvious changes in diaphragm atrophy, but the contractiity of the diaphragm decreased, which was consistent with our previous research results. The mRNA expression levels of the above genes in sepsis group were significantly increased (P<0.05), and were significantly decreased after Calpeptin pretreatment(P<0.05) by qRT-PCR. Conclusion: Diaphragm muscles are prone to dysfunction during sepsis. Calpeptin, a calpain inhibitor, can significantly reduce the activation of inflammation, apoptosis and autophagy pathways induced by LPS. |
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