尼克酰胺对内毒素血症及脓毒症小鼠的治疗作用

发布时间：2018-03-18 11:26

本文选题：尼克酰胺　切入点：内毒素血症　出处：《重庆医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：通过研究证实尼克酰胺（NAM）在小鼠内毒素血症及脓毒症模型中的治疗作用，为临床治疗脓毒症提供新的策略。方法：腹腔注射脂多糖（10ug/kg）联合半乳糖胺（700mg/kg）（LPS/D-Gal）建立小鼠内毒素所致肝损伤模型，腹腔注射致死剂量脂多糖（20mg/kg）建立内毒素血症模型，盲肠结扎穿孔（CLP）建立多种微生物感染的小鼠脓毒症模型。LPS/D-Gal诱导的肝损伤模型中，分别于建模前30min腹腔注射不同剂量的NAM（100mg/kg，200mg/kg，400mg/kg）或建模后1h、3h、6h腹腔注射NAM（400mg/kg），观察小鼠的存活情况并作生存曲线。于建模后6h，麻醉小鼠，摘眼球取血，检测血清中转氨酶水平，一并取肝组织，半胱氨酸天冬氨酸蛋白酶（Caspase）活性检测及TUNEL检测观察肝细胞凋亡，肝组织病理切片观察形态学改变；于建模后1.5h取血检测血清炎症因子TNF-α水平。致死性内毒素血症模型中，建模前30min或建模后1h给予NAM（400mg/kg，i.p.），观察小鼠存活情况并作生存曲线。建模后3h取血，测血清TNF-α、IL-6水平，建模后12h同样方法取血，测血清ALT及BUN水平，并取肺组织，测肺干湿重比，肺组织病理切片用于观察形态学改变。CLP模型中，于手术后1h给予NAM（400mg/kg，i.p.），观察小鼠存活情况并作生存曲线。结果：在LPS/D-Gal诱导的肝损伤模型中，NAM显著降低血清转氨酶水平，并减轻肝组织损伤。NAM还降低肿瘤坏死因子TNF-α水平。通过TUNEL染色试验及半胱氨酸天冬氨酸蛋白酶（Caspase）活性检测，证实NAM可以减少肝细胞凋亡。此外，生存分析显示NAM可降低LPS/D-Gal诱导小鼠的死亡率。在致死性内毒素血症中， NAM降低血清促炎因子水平及多器官功能损伤，，降低肺组织干湿重比，减轻血清转氨酶水平及血尿素氮水平。生存分析中，NAM提高了致死性内毒素血症及CLP脓毒症小鼠的存活率。结论：研究证实，NAM减轻炎性损伤并提高小鼠存活率，对脓毒症小鼠可能发挥治疗作用。
[Abstract]:Objective: to investigate the therapeutic effect of nicotinamide (NAM) on endotoxemia and sepsis in mice. Methods: lipopolysaccharide (LPS) and galactosamine (700 mg / kg) were injected intraperitoneally to establish lipopolysaccharide induced liver injury model in mice, and lipopolysaccharide (LPS) 20 mg / kg intraperitoneal injection was used to establish endotoxemia model. Cecal ligation and perforation (CLP) was used to establish the sepsis model of mice infected by various microbes. LPS- D-Gal induced liver injury model. At 30 minutes before modeling, different doses of NAMN 100 mg / kg were injected intraperitoneally with 200 mg / kg of NAMN (400 mg / kg), or 1 hour after modeling, 400 mg / kg NAMN was injected intraperitoneally for 6 h after modeling. The survival of the mice was observed and the survival curve was made. At 6 h after modeling, the anesthetized mice were anesthetized, the blood was taken from eyeball, the level of serum aminotransferase was detected, and the liver tissue was taken. The activity of cysteine aspartate proteinase (caspase) and the detection of TUNEL were used to observe the apoptosis of hepatocytes, the pathological sections of liver were observed for morphological changes, the serum levels of inflammatory factor TNF- 伪 were measured at 1.5 h after modeling, and the levels of TNF- 伪 were detected in the model of fatal endotoxemia. 30 minutes before modeling or 1 hour after modeling, the survival of mice was observed and the survival curve was made. Blood samples were taken at 3 hours after modeling, serum TNF- 伪 IL-6 levels were measured, serum ALT and BUN levels were measured at 12 hours after modeling, lung tissue was taken and lung dry-wet weight ratio was measured. Lung histopathological sections were used to observe the morphological changes in the model of LPS/D-Gal. The rats were treated with NAMN 400 mg / kg i.p. at 1 h after operation. Results: in the model of liver injury induced by LPS/D-Gal, NAM significantly decreased the level of serum aminotransferase. NAM also reduced the level of TNF- 伪 in liver tissue. By TUNEL staining and caspase activity assay, it was proved that NAM could reduce the apoptosis of hepatocytes. Survival analysis showed that NAM could reduce the mortality of mice induced by LPS/D-Gal. In fatal endotoxemia, NAM decreased the level of serum pro-inflammatory factor and the damage of multi-organ function, and decreased the ratio of dry to wet weight of lung tissue. In survival analysis, nam increased the survival rate of mice with fatal endotoxemia and CLP sepsis. It may play a therapeutic role in sepsis mice.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R459.7

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