D-柠檬烯对酒精性脑损伤大鼠NF-κB、IκB及TNF-α、IL-10表达的影响

发布时间：2018-04-05 15:26

本文选题：D-柠檬烯　切入点：酒精性脑损伤　出处：《青岛大学》2015年硕士论文

【摘要】：目的:观察D-柠檬烯对酒精性脑损伤大鼠脑组织核因子-κB(NF-κB)、核因子κB抑制蛋白(IκB)表达及与肿瘤坏死因子(TNF-α)、白介素-10(IL-10)水平的影响。方法:60只雄性SPF级Wistar大鼠,体重为180~200g。适应性喂养7天后,按随机数字表法,将其分为6组(n=10)。A组:空白对照组,每日予以生理盐水灌胃前2周为8ml/(kg·d),后4周为12ml/(kg·d)。B组:酒精模型组,每日予以50%(V/V)乙醇灌胃,前2周为8ml/(kg·d),后4周为12ml/(kg·d)。C、D、E三组分别给予不同浓度的D-柠檬烯灌胃,剂量分别为100、200、400mg/(kg·d),同时每日给予50%(V/V)酒精,剂量同模型组。F组为单独D-柠檬烯高剂量组,每日给予D-柠檬烯的量为400mg/(kg·d)。末次给药后,大鼠禁食不禁水12 h,10%水合氯醛麻醉,留取脑组织。左侧脑组织用HE染色法观察其病理学改变;免疫组织化学法检测脑组织中NF-κB、IκB表达情况,计算其阳性细胞数;右侧脑组织用酶联免疫吸附法(ELISA)检测脑匀浆中TNF-α、IL-10的浓度。结果:(1)组织病理学观察:经过HE染色后,观察发现酒精组海马区神经元数量减少,分布不均匀,细胞核明显固缩变形,难以分辨。空白对照组及单独D-柠檬烯高剂量组海马区神经元排列层次较清,细胞结构完整(核圆而大、核膜清楚、HE染色均匀),个别神经元出现异形,表现为核固缩、胞体比正常缩小、形状不规则、胞质染色为深红色。柠檬烯各剂量组海马区有数量不等的异形神经元,与酒精模型组相比,异形神经元数量明显减少。(2)NF-κB、IκB蛋白在大鼠脑组织中的阳性细胞数:酒精模型组与空白对照组相比NF-κB阳性细胞数增多,IκB阳性细胞数减少(P0.05);给予不同剂量的柠檬烯后各组大鼠脑组织中的NF-κB阳性细胞数随着柠檬烯剂量的增加逐渐减少、IκB阳性细胞数逐渐增多,同模型组对比(P0.05);空白对照组与单纯D-柠檬烯药物组之间差异无统计学意义。(3)大鼠脑组织匀浆中TNF-α的表达:模型组脑匀浆中TNF-α的表达明显高于对照组(P0.05);柠檬烯低、中、高剂量组大鼠脑匀浆中TNF-α表达降低,与模型组相比(P0.05);单纯D-柠檬药物组与空白对照组相比差异无统计学意义。(4)大鼠脑组织匀浆中IL-10的表达:酒精模型组中IL-10的表达高于空白对照组(P0.05);柠檬烯中、高剂量组大鼠脑匀浆IL-10的表达明显高于酒精模型组(P0.05);空白对照组与单纯D-柠檬药物组相比差异无统计学意义。结论:D-柠檬烯可以减轻过量酒精摄入引起的脑组织损伤,作用机制可能与抑制NF-κB的表达、上调IκB的表达以及调节TNF-α、IL-10的水平有关。
[Abstract]:Aim: to observe the effects of D-limonene on the expression of nuclear factor- 魏 B (NF- 魏 B), nuclear factor- 魏 B inhibitor (NF-kB) and the levels of tumor necrosis factor TNF- 伪 (TNF- 伪) and interleukin-10 (IL-10) in brain tissue of rats with alcoholic brain injury.Methods 60 male Wistar rats of SPF grade, weighing 180 ~ 200g.After the last administration, rats were anesthetized with 10% chloral hydrate for 12 h after fasting.The pathological changes of left brain were observed by HE staining, the expression of NF- 魏 B I 魏 B and the number of positive cells were measured by immunohistochemical method, and the concentration of TNF- 伪 IL-10 in brain homogenate was detected by enzyme linked immunosorbent assay (Elisa).Results (1) histopathological observation: after HE staining, it was found that the number of hippocampal neurons in the alcohol group was decreased, the distribution of neurons was uneven, the nucleus was obviously constricted and deformed, and it was difficult to distinguish between them.The hippocampal neurons in the blank control group and the high-dose D- limonene group were well arranged, and the structure of the neurons was intact (the nucleus was round and large, the nuclear membrane was clear and HE staining was uniform), some neurons showed abnormal shape, which showed nuclear pyknosis, and the cell body was smaller than normal.The shape was irregular and the cytoplasm stained crimson.Different doses of limonene had different numbers of hippocampal neurons, compared with the alcohol model group.The number of NF- 魏 B positive cells in the brain tissue of rats was significantly decreased. The number of NF- 魏 B positive cells in the alcohol model group was higher than that in the blank control group, and the number of I 魏 B positive cells in the alcohol model group was lower than that in the blank control group (P 0.05), and the number of I 魏 B positive cells in different doses of limonene was significantly lower than that in the control group (P 0.05).The number of NF- 魏 B positive cells decreased with the increase of limonene dose, and the number of I 魏 B positive cells increased with the increase of limonene dose.The expression of TNF- 伪 in brain homogenate of rats in high dose group was decreased.Compared with the model group, the expression of IL-10 in the brain homogenate of rats was higher in the alcohol model group than that in the blank control group, and in limonene, the expression of IL-10 was higher in the alcohol model group than in the control group.The expression of IL-10 in brain homogenate of rats in high dose group was significantly higher than that in alcohol model group (P 0.05), but there was no significant difference between blank control group and D- lemon drug group.Conclusion the brain tissue injury induced by excessive alcohol intake could be alleviated by 1: D- limonene. The mechanism may be related to the inhibition of NF- 魏 B expression, the up-regulation of I 魏 B expression and the regulation of TNF- 伪 -IL-10 level.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R595.6;R747.9

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