参附注射液对内毒素休克模型大鼠肺组织炎症的改善作用及抗炎机制研究
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篇名: 参附注射液对内毒素休克模型大鼠肺组织炎症的改善作用及抗炎机制研究
TITLE:
摘要: 目的:研究参附注射液对内毒素休克模型大鼠肺组织炎症的改善作用及其抗炎机制。方法:将48只大鼠随机分为正常组、模型组、地塞米松组(阳性对照,1 mg/kg)和参附注射液低、中、高剂量组(5、10、15 mL/kg),每组8只。除正常组外,其余各组大鼠腹腔注射脂多糖(LPS)复制内毒素休克模型,造模后各组大鼠腹腔注射给药1次。给药24 h后,苏木精-伊红(HE)染色后观察大鼠肺组织病理学变化并进行病理学评分;逆转录-聚合酶链式反应(RT-PCR)法测定肺组织中核转录因子κB(NF-κB)信号通路相关蛋白P65、P50 mRNA的表达水平;Western blot法测定肺组织中P65、P50蛋白的表达水平,并检测肺组织细胞核和细胞浆中P65蛋白的表达水平;采用酶联免疫吸附试验(ELISA)检测大鼠血浆中肿瘤坏死因子α(TNF-α)水平。结果:与正常组比较,模型组大鼠肺泡间隔增厚,血管明显充血,肺间质大量中性粒细胞浸润;肺组织病理学评分和P65、P50 mRNA及蛋白的表达水平均显著升高(P<0.01或P<0.001),细胞核、细胞浆中P65蛋白和血浆中TNF-α水平均显著升高(P<0.001)。与模型组比较,地塞米松组和参附注射液中、高剂量组大鼠肺泡结构完整,无明显出血,炎性细胞浸润程度明显改善;肺组织病理学评分和P65、P50 mRNA及蛋白的表达水平以及血浆中TNF-α水平均显著降低(P<0.05或P<0.01或P<0.001),地塞米松组和参附注射液低、中剂量组大鼠肺组织细胞核、细胞浆中P65蛋白表达水平均显著降低(P<0.05或P<0.01或P<0.001)。结论:参附注射液可通过降低肺组织中P65、P50 mRNA及蛋白的表达水平及血浆中TNF-α的水平,改善内毒素休克模型大鼠肺组织的炎症反应。
ABSTRACT: OBJECTIVE: To study the improvement and anti-inflammation mechanism of Shenfu injection on lung tissue of endotoxin shock model rats. METHODS: Totally 48 rats were randomized into control group,model group,dexamethasone group (positive control,1 mg/kg) and Shenfu injection low-dose,medium-dose and high-dose groups (5,10,15 mL/kg),with 8 rats in each group. Except for normal group, other groups were given intraperitoneal injection of lipopolysaccharide (LPS) to induce endotoxin shock model. After modeling, each group was given relevant medicine once intraperitoneally. 24 h after medication, HE staining was used to observe pathological changes of lung tissue in rats and pathological scoring was conducted. RT-PCR was used to determine mRNA levels of P65 and P50 proteins related to NF-κB signaling pathway. Western blot assay was used to determine the expression levels of P65 and P50 proteins in lung tissue, and the expression levels of P65 protein in nucleus and cytoplasm of lung tissue were also determined. The level of TNF-α in plasma in rats were determined by ELISA. RESULTS: Compared with control group, alveolar septum became thicker, obvious vascular engorgement was found, and a large number of neutrophils infiltrated the interstitium in model group. Histopathological score, mRNA and protein expression levels of P65 and P50 in lung tissues were increased significantly (P<0.01 or P<0.001); the protein expression of levels P65 in nucleus and cytoplasm and level of TNF-α in plasma were increased significantly (P<0.001). Compared with model group, alveolar structure of rats in dexamethasone group and Shenfu injection medium-dose and high-dose groups was complete, no obvious bleeding was observed, and the degree of inflammatory cell infiltration was improved significantly. Histopathological score, mRNA and protein expression levels of P65 and P50 in lung tissue and level of TNF-α in plasma were decreased significantly (P<0.05 or P<0.01 or P<0.001). The protein expression level of P65 in nucleus and cytoplasm of lung tissue were decreased significantly in dexamethasone group and Shenfu injection low-dose and medium-dose groups were decreased significantly (P<0.05 or P<0.01 or P<0.001). CONCLUSIONS: Shenfu injection can decrease mRNA and protein expression levels of P65 and P50 in lung tissue, level of TNF-α in plasma, and protect lung tissue of endotoxin shock rats.
期刊: 2019年第30卷第11期
作者: 刘霞,艾飞,褚春薇,陈向云,郭俊峰,杨毅,梅丽艳,苗纪飞,温泉,叶森,黎晖
AUTHORS: LIU Xia,AI Fei,CHU Chunwei,CHEN Xiangyun,GUO Junfeng,YANG Yi,MEI Liyan,MIAO Jifei,WEN Quan,YE Sen,LI Hui
关键字: 参附注射液;急性肺损伤;内毒素休克;核转录因子κB;P65;P50;大鼠
KEYWORDS: Shenfu injection; Acute lung injury; Endotoxin shock; NF-κB; P65; P50; Rat
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