小檗碱预防脂多糖性肝损伤的机制研究

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目的:观察小檗碱对脂多糖性肝损伤的防治效果及其作用机制。方法:将雄性昆明小鼠随机分成4组:①对照组:用蒸馏水灌胃(0.01 mL/g),每天1次,共5 d,第5 d灌胃后1 h,腹腔注射生理盐水(0.02 mL/g);②小檗碱组:用5 g/L中性硫酸小檗碱灌胃(0.01 mL/g),每天1次,共5 d,第5 d灌胃后1 h,腹腔注射生理盐水(0.02mL/g);③脂多糖(LPS)组:除腹腔注射LPS(0.02 mL/g,28 mg/kg)外,其余处理同①;④小檗碱防治组:除腹腔注射LPS(0.02 mL/g,28 mg/kg)外,其余处理同②。腹腔注射后2 h和10 h去眼球取血,分别检测各组小鼠血清中TNF-α的含量以及丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)的活性;另于腹腔注射后24 h取肝组织标本,观察各组小鼠肝脏的组织学和超微结构的变化,同时测定肝组织MDA的含量及SOD的活性。结果:LPS组小鼠血清ALT、AST活性明显高于对照组(P<0.01),而小檗碱防治组小鼠血清ALT和AST活性明显低于LPS组(P<0.05)。腹腔注射LPS后2 h,LPS组小鼠血清中TNF-α含量明显高于小檗碱防治组。组织学检查发现,LPS组小鼠肝细胞水肿、变性、坏死,肝窦充血;电镜下可见,肝细胞核溶解、部分核膜不完整,线粒体肿胀、嵴消失。小檗碱防治组小鼠肝脏的病理变化明显轻于LPS组。此外,LPS组肝脏组织中MDA的含量明显高于对照组(P<0.01),而小檗碱防治组肝脏组织中MDA的含量低于LPS组(P<0.05),但小檗碱防治组肝组织中SOD活性与LPS组比较无显著差异。结论:小檗碱可以减轻LPS引起的肝脏损伤,其作用机制可能与其抑制LPS诱导的TNF-α释放,减少肝组织脂质过氧化和保护肝细胞线粒体有关。 Objective: To observe the effect of berberine on lipopolysaccharide-induced liver injury and its mechanism of action. METHODS: Male Kunming mice were randomly divided into 4 groups: 1 control group: intragastrically administered distilled water (0.01 mL/g) once daily for 5 days, and intraperitoneal injection of physiological saline (0.02 mg/d) on the 5th day after intragastric administration. mL/g);2 Berberine group: intragastrically administered with 5 g/L neutral berberine sulfate (0.01 mL/g) once a day for 5 days, intraperitoneal injection 1 h after 5 d intragastric administration Physiological saline (0.02mL/g); 3Lipopolysaccharide (LPS) group: In addition to LPS (0.02 mL/g, 28 mg/kg), the rest were treated with 1; 4 berberine prevention group: Intraperitoneal injection of LPS (0.02 mL/g, 28 mg/kg), the remaining treatment is the same as 2. Two hours and 10 hours after intraperitoneal injection, the blood was collected from the eyeball and the levels of serum TNF-α and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity were measured in each group. In addition, liver specimens were taken 24 h after intraperitoneal injection to observe the histological and ultrastructural changes of the livers of the mice in each group. Simultaneously, the content of MDA in liver tissues and the activity of SOD were measured. RESULTS: Serum ALT and AST activity in LPS group was significantly higher than that in control group (P<0.01). Serum ALT and AST activity in mice treated with berberine was significantly lower than that in LPS group (P<0.05). Two hours after intraperitoneal injection of LPS, the serum level of TNF-α in LPS group was significantly higher than that of berberine prevention group. Histological examination revealed that mice in the LPS group had edema, degeneration, necrosis, and hepatic sinusoidal congestion; electron microscopic examination revealed that the hepatocyte nucleus was lysed, part of the nuclear membrane was incomplete, and the mitochondria were swollen and the iliac crest disappeared. The pathological changes of liver in mice treated with berberine were significantly lighter than those in LPS group. In addition, the content of MDA in liver tissue of LPS group was significantly higher than that of control group (P<0.01), while the content of MDA in liver tissue of berberine prevention group was lower than that of LPS group (P<0.05), but berberine prevented the liver of control group. There was no significant difference in SOD activity between the tissues and the LPS group. Conclusion: Berberine can reduce liver injury induced by LPS. Its mechanism may be related to its inhibition of LPS-induced TNF-α release, reduction of liver lipid peroxidation and protection of hepatocyte mitochondria.
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