树莓茎对药物性肝损伤小鼠肝组织HMGB1、TLR4、NF-κB蛋白表达的影响

发布时间：2018-10-31 08:26

【摘要】：目的:研究树莓茎抗药物性肝损伤的作用及对肝组织中HMGB1、TLR4、NF-κB蛋白表达的影响,为树莓茎的药用开发和临床应用提供实验依据。方法:72只健康雄性ICR小鼠,随机分成6组,每组12只,即空白组、模型组、联苯双酯组、树莓茎低剂量组、树莓茎中剂量组、树莓茎高剂量组。树莓茎低、中、高剂量组分别给予树莓茎水煎液提取物进行灌胃,剂量分别为1.14g/kg、2.28g/kg、4.56g/kg,联苯双酯组给予联苯双酯混悬液0.15g/kg,空白组、模型组给予等体积的生理盐水,给药体积为20mL/kg。每日灌胃一次,连续四周。末次给药1小时后,除空白组以外,其余各组ICR小鼠给予对乙酰氨基酚500mg/kg腹腔注射进行造模,并观察小鼠一般状态,禁食不禁水。造模24小时后小鼠眼球取血,检测小鼠血清中ALT、AST的水平;取一部分肝脏用于HE染色并观察其病理变化;ELISA法测定小鼠肝组织IL-1β、TNF-α的含量;免疫组织化学方法测定小鼠肝组织中HMGB1、TLR4、NF-κB蛋白的表达情况。结果:与空白组相比较,模型组小鼠血清中ALT、AST水平明显升高,差异具有显著性,有统计学意义(P0.01);与模型组相比较,树莓茎高、中、低剂量组能不同程度的降低药物性肝损伤小鼠血清中ALT、AST的水平,有显著性差异,具有统计学意义(P0.01);HE染色结果可见树莓茎高、中剂量组明显减轻对乙酰氨基酚诱导小鼠肝细胞的肿胀程度和炎性细胞浸润的坏死面积;ELISA法检测结果发现树莓茎可降低小鼠肝组织IL-1β、TNF-α的含量(P0.01,P0.05);免疫组化检测结果发现树莓茎可降低小鼠肝组织中HMGB1、TLR4、NF-κB蛋白的表达(P0.01,P0.05)。结论:树莓茎可通过抑制小鼠肝组织中HMGB1、TLR4、NF-κB蛋白的表达,调控炎症细胞因子的水平,起到抗药物性肝损伤的作用。
[Abstract]:Objective: to study the effects of raspberry stem on drug-induced liver injury and the expression of HMGB1,TLR4,NF- 魏 B protein in liver tissue, and to provide experimental evidence for the medicinal development and clinical application of raspberry stem. Methods: 72 healthy male ICR mice were randomly divided into 6 groups with 12 in each group: blank group, model group, biphenyl diester group, low dose group of raspberry stem, middle dose group of stem of raspberry, high dose group of stem of raspberry. The low, middle and high doses of raspberry stem extract were given by gavage, the dose was 1.14 g / kg ~ (2. 28) g 路kg ~ (-1) 路kg ~ (-1), the biphenyl diester group was given 0.15 g / kg / kg biphenyl diester suspension, and the blank group was given biphenyl diester suspension 0.15 g / kg, respectively. The model group was given the same volume of normal saline and 20 mL / kg of saline. Gavage once a day for four weeks. One hour after the last administration, the other ICR mice were given paracetamol 500mg/kg intraperitoneally to model the mice except the blank group. The general state of the mice was observed and the water was fasted. After 24 hours of model, blood was taken from eyeball of mice to detect the level of ALT,AST in serum, part of liver was used for HE staining and pathological changes were observed, content of IL-1 尾 and TNF- 伪 in liver tissue of mice was determined by ELISA method. The expression of HMGB1,TLR4,NF- 魏 B protein in mouse liver was detected by immunohistochemical method. Results: compared with the blank group, the serum ALT,AST level in the model group was significantly higher than that in the blank group (P0.01). Compared with the model group, the Raspberry stem height, medium and low dose group can reduce the serum ALT,AST level of the drug-induced liver injury mice in varying degrees, there is significant difference (P0.01); The results of HE staining showed that the stem of raspberry was high, and the swelling degree of hepatocytes induced by acetaminophen and the necrotic area of inflammatory cells were significantly reduced in middle dose group. The results of ELISA assay showed that the stem of raspberry could reduce the contents of IL-1 尾 and TNF- 伪 in liver tissue of mice (P 0.01 P 0.05). The results of immunohistochemistry showed that the stem of raspberry could reduce the expression of HMGB1,TLR4,NF- 魏 B protein in liver tissue of mice (P0.01P05). Conclusion: the stem of raspberry can inhibit the expression of HMGB1,TLR4,NF- 魏 B protein and regulate the level of inflammatory cytokines in the liver tissue of mice.
【学位授予单位】：黑龙江中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5

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