【摘要】：目的：本研究從氧化應(yīng)激為入口,觀察不同時(shí)間窗下具有養(yǎng)陰益氣、活血化瘀功效的參芪復(fù)方對糖尿病大血管病變GK大鼠血糖、血清CHOL、8-iso-PGF2α;胸主動(dòng)脈勻漿SOD、MDA;胸主動(dòng)脈病理形態(tài)、細(xì)胞凋亡的影響,進(jìn)行量-時(shí)-效關(guān)系分析,推測其可能通過抑制氧化應(yīng)激、阻斷“代謝記憶”途徑來保護(hù)大血管。并應(yīng)用基因芯片技術(shù)尋找參芪復(fù)方防治糖尿病大血管病變的分子靶點(diǎn),探討?zhàn)B陰益氣活血法阻斷“代謝記憶”的生物學(xué)作用及其機(jī)理,為臨床推廣中醫(yī)藥防治糖尿病大血管病變提供理論依據(jù)。方法：160只GK大鼠適應(yīng)性飼養(yǎng)1周后,依據(jù)隨機(jī)血糖水平由高到低排列編號,隨機(jī)分為模型組、參芪復(fù)方高劑量組(參高組)、參芪復(fù)方中劑量組(參中組)、參芪復(fù)方低劑量組(參低組)、二甲雙胍組(西藥組),每組32只,給予高脂飼料飼養(yǎng)；Wistar組(空白組)32只給予普通飼料飼養(yǎng)；連續(xù)喂養(yǎng)12周,建立糖尿病“代謝記憶”模型。造模完成后,模型組和空白組給予5ml/kg.d生理鹽水灌服,參高組給予2.88g/kg.d參芪復(fù)方浸膏灌服,參中組給予1.44g/kg.d參芪復(fù)方浸膏灌服,參低組給予0.72g/kg.d參芪復(fù)方浸膏灌服,西藥組給予0.1g/kg.d二甲雙胍灌服,治療干預(yù)16周。實(shí)驗(yàn)期間從整體水平觀察各組大鼠的一般情況,監(jiān)測空腹血糖變化；各組大鼠分別在灌胃后第8、16周末處死,檢測血清CHOL、胸主動(dòng)脈勻漿SOD、MDA等指標(biāo)；從組織水平,檢測胸主動(dòng)脈細(xì)胞凋亡情況,HE染色對胸主動(dòng)脈進(jìn)行形態(tài)學(xué)觀察；觀察不同時(shí)間窗下,不同干預(yù)方式對模型大鼠“代謝記憶”的影響效果。借助基因芯片技術(shù)對胸主動(dòng)脈組織進(jìn)行基因表達(dá)譜檢測,篩選出差異表達(dá)基因,進(jìn)行GO注釋,Pathway分析,解釋生物學(xué)意義。結(jié)果：參中組的作用優(yōu)于參高組和參低組,能夠明顯改善GK大鼠的一般狀況、血糖、血脂、氧化應(yīng)激指標(biāo)、胸主動(dòng)脈病理形態(tài)及細(xì)胞凋亡。參中組與模型組差異表達(dá)基因,經(jīng)GO注釋、Pathway分析,主要涉及大分子代謝過程、細(xì)胞死亡、信號轉(zhuǎn)導(dǎo)、細(xì)胞遷移和MAPK信號通路、刺猬信號轉(zhuǎn)導(dǎo)通路、Notch信號通路、胰島素信號轉(zhuǎn)導(dǎo)等通路：涉及到Csnk1d、Lama4、Elavl1、Abi1、Anxa1、 Nsg1等基因。結(jié)論：參中組可以更好地改善糖尿病大血管病變GK大鼠的一般狀態(tài),降低血糖,改善脂代謝紊亂,減輕氧化應(yīng)激,保護(hù)大鼠血管內(nèi)皮損傷,阻斷“代謝記憶”。其作用機(jī)理可能是通過調(diào)控大分子代謝過程、細(xì)胞死亡、信號轉(zhuǎn)導(dǎo)、細(xì)胞遷移和MAPK信號通路、刺猬信號轉(zhuǎn)導(dǎo)通路、Notch信號通路、胰島素信號轉(zhuǎn)導(dǎo)等通路及Csnk1d、Lama4、Elav11、Abi1、AnxaK1、Nsg1等基因,促進(jìn)糖尿病大血管病變早期受損血管內(nèi)皮細(xì)胞修復(fù),改善氧化應(yīng)激對血管的損害。其中,參芪復(fù)方可能通過調(diào)控MAPK這條重要的信號通路,減輕氧化應(yīng)激,抑制血管內(nèi)皮細(xì)胞凋亡,消除炎癥反應(yīng),阻斷糖尿病大血管病變“代謝記憶”,從而防治糖尿病大血管病變。
[Abstract]:Objective: to observe the effects of Shenqi compound on blood sugar, serum CHOL,8-iso-PGF2 偽 and thoracic aortic homogenate SOD,MDA; in GK rats with diabetic vascular disease under different time window, which can nourish yin and invigorate qi and promote blood circulation and remove blood stasis. The effects of pathological morphology and apoptosis of thoracic aorta were analyzed by dose-time-effect relationship. It was speculated that the aorta might protect large vessels by inhibiting oxidative stress and blocking "metabolic memory" pathway. The molecular target of Shenqi compound for prevention and treatment of diabetic macrovascular disease was found by using gene chip technology, and the biological effect and mechanism of the method of nourishing yin, supplementing qi and activating blood circulation to block "metabolic memory" were discussed. To provide a theoretical basis for the prevention and treatment of diabetic macroangiopathy with traditional Chinese medicine. Methods: one week after adaptive feeding, 160 GK rats were randomly divided into model group, Shenqi compound high dose group and Shenqi compound medium dose group according to the random blood glucose level from high to low. Shenqi compound low-dose group (Shen-low group), metformin group (western medicine group), 32 rats in each group, were fed with high-fat diet; Wistar group (blank group) was fed with normal diet for 12 weeks to establish diabetes "metabolic memory" model. After the model was made, the model group and blank group were given 5ml/kg.d saline, the Shengao group was given 2.88g/kg.d Shenqi compound extract, and the Shenzhong group was given 1.44g/kg.d Shenqi compound extract. Shen-low group was given 0.72g/kg.d Shenqi compound extract and western medicine group was given 0.1g/kg.d metformin for 16 weeks. During the experiment, the general situation of the rats in each group was observed at the whole level, and the changes of fasting blood glucose were monitored. The rats in each group were killed at the end of 816 weeks after gavage, and the serum SOD,MDA of thoracic aorta homogenate of CHOL, were detected. The apoptosis of thoracic aorta was detected at the tissue level, the morphology of thoracic aorta was observed by HE staining, and the effect of different intervention methods on "metabolic memory" in model rats was observed under different time windows. The gene expression profiles of thoracic aorta were detected by gene chip technique. The differentially expressed genes were screened out, and the biological significance was explained by GO annotation and Pathway analysis. Results: the effect of Shenzhong group was better than that of Shengao group and Shen-low group, and could obviously improve the general condition, blood glucose, blood lipid, oxidative stress index, pathological morphology of thoracic aorta and apoptosis of GK rats. By GO annotation and Pathway analysis, the differentially expressed genes in Shenzhong group and model group were mainly involved in macromolecular metabolic process, cell death, signal transduction, cell migration and MAPK signaling pathway, hedgehog signal transduction pathway and Notch signal pathway. Insulin signal transduction pathway: involved in Csnk1d,Lama4,Elavl1,Abi1,Anxa1, Nsg1 and other genes. Conclusion: Shenzhong group can better improve the general state of GK rats with diabetic macrovascular disease, decrease blood glucose, improve lipid metabolism disorder, alleviate oxidative stress, protect vascular endothelial injury and block "metabolic memory". Its mechanism may be by regulating macromolecular metabolic process, cell death, signal transduction, cell migration and MAPK signaling pathway, hedgehog signal transduction pathway, Notch signal pathway, insulin signal transduction pathway and Csnk1d,Lama4,Elav11,Abi1,AnxaK1,. Nsg1 and other genes can promote the repair of vascular endothelial cells in the early stage of diabetic macrovascular disease and improve the oxidative stress damage to the blood vessels. Among them, Shenqi compound may reduce oxidative stress, inhibit apoptosis of vascular endothelial cells, eliminate inflammatory reaction and block "metabolic memory" of diabetic macrovascular disease by regulating the important signal pathway of MAPK. In order to prevent and treat diabetic macrovascular disease.
【學(xué)位授予單位】：成都中醫(yī)藥大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R259
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本文編號：2300459