本文選題：D-檸檬烯　切入點：酒精性腦損傷　出處：《青島大學(xué)》2015年碩士論文

【摘要】：目的:觀察D-檸檬烯對酒精性腦損傷大鼠腦組織核因子-κB(NF-κB)、核因子κB抑制蛋白(IκB)表達及與腫瘤壞死因子(TNF-α)、白介素-10(IL-10)水平的影響。方法:60只雄性SPF級Wistar大鼠,體重為180~200g。適應(yīng)性喂養(yǎng)7天后,按隨機數(shù)字表法,將其分為6組(n=10)。A組:空白對照組,每日予以生理鹽水灌胃前2周為8ml/(kg·d),后4周為12ml/(kg·d)。B組:酒精模型組,每日予以50%(V/V)乙醇灌胃,前2周為8ml/(kg·d),后4周為12ml/(kg·d)。C、D、E三組分別給予不同濃度的D-檸檬烯灌胃,劑量分別為100、200、400mg/(kg·d),同時每日給予50%(V/V)酒精,劑量同模型組。F組為單獨D-檸檬烯高劑量組,每日給予D-檸檬烯的量為400mg/(kg·d)。末次給藥后,大鼠禁食不禁水12 h,10%水合氯醛麻醉,留取腦組織。左側(cè)腦組織用HE染色法觀察其病理學(xué)改變;免疫組織化學(xué)法檢測腦組織中NF-κB、IκB表達情況,計算其陽性細胞數(shù);右側(cè)腦組織用酶聯(lián)免疫吸附法(ELISA)檢測腦勻漿中TNF-α、IL-10的濃度。結(jié)果:(1)組織病理學(xué)觀察:經(jīng)過HE染色后,觀察發(fā)現(xiàn)酒精組海馬區(qū)神經(jīng)元數(shù)量減少,分布不均勻,細胞核明顯固縮變形,難以分辨。空白對照組及單獨D-檸檬烯高劑量組海馬區(qū)神經(jīng)元排列層次較清,細胞結(jié)構(gòu)完整(核圓而大、核膜清楚、HE染色均勻),個別神經(jīng)元出現(xiàn)異形,表現(xiàn)為核固縮、胞體比正�？s小、形狀不規(guī)則、胞質(zhì)染色為深紅色。檸檬烯各劑量組海馬區(qū)有數(shù)量不等的異形神經(jīng)元,與酒精模型組相比,異形神經(jīng)元數(shù)量明顯減少。(2)NF-κB、IκB蛋白在大鼠腦組織中的陽性細胞數(shù):酒精模型組與空白對照組相比NF-κB陽性細胞數(shù)增多,IκB陽性細胞數(shù)減少(P0.05);給予不同劑量的檸檬烯后各組大鼠腦組織中的NF-κB陽性細胞數(shù)隨著檸檬烯劑量的增加逐漸減少、IκB陽性細胞數(shù)逐漸增多,同模型組對比(P0.05);空白對照組與單純D-檸檬烯藥物組之間差異無統(tǒng)計學(xué)意義。(3)大鼠腦組織勻漿中TNF-α的表達:模型組腦勻漿中TNF-α的表達明顯高于對照組(P0.05);檸檬烯低、中、高劑量組大鼠腦勻漿中TNF-α表達降低,與模型組相比(P0.05);單純D-檸檬藥物組與空白對照組相比差異無統(tǒng)計學(xué)意義。(4)大鼠腦組織勻漿中IL-10的表達:酒精模型組中IL-10的表達高于空白對照組(P0.05);檸檬烯中、高劑量組大鼠腦勻漿IL-10的表達明顯高于酒精模型組(P0.05);空白對照組與單純D-檸檬藥物組相比差異無統(tǒng)計學(xué)意義。結(jié)論:D-檸檬烯可以減輕過量酒精攝入引起的腦組織損傷,作用機制可能與抑制NF-κB的表達、上調(diào)IκB的表達以及調(diào)節(jié)TNF-α、IL-10的水平有關(guān)。
[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- 偽 (TNF- 偽) 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- 偽 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- 偽 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- 偽 -IL-10 level.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R595.6;R747.9

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