【摘要】：目的應(yīng)用高糖高脂飼料喂養(yǎng)聯(lián)合小劑量鏈脲佐菌素(Streptozotocin,STZ)腹腔注射構(gòu)建大鼠2型糖尿病模型,觀察糖尿病狀態(tài)下大鼠主動(dòng)脈血管組織中高遷移率族蛋白A1(high mobility group A1,HMGA1)m RNA和蛋白的表達(dá)水平,并探討可能影響其變化的病理因素,試圖為糖尿病血管并發(fā)癥的發(fā)病機(jī)制及防治研究提供新的方向。方法4周齡雄性SD大鼠按隨機(jī)分組方法分為:正常對(duì)照組(normol contol group,NC)、高糖高脂喂養(yǎng)組(high-sucrose-high-fat diets group,HSFDs)、糖尿病造模組(diabetes mellitus group,DM)。NC組懫用普通標(biāo)準(zhǔn)大鼠飼料喂養(yǎng),HSFDs組及DM組均懫用高糖高脂飲食喂養(yǎng),起始及喂養(yǎng)4周后分別測(cè)量體重(body weight,BW)并檢測(cè)空腹血糖(fasting blood glucose,FBG)、非空腹血糖(nonfasting blood glucose,NFBG)、血清總膽固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白(low density lipoprotein,LDL)、高密度脂蛋白(high density lipoprotein,HDL)及空腹胰島素(fasting insulin,FINS)等相關(guān)生化指標(biāo)的水平。4周末,DM組大鼠腹腔注射STZ(30mg/Kg),3天后開(kāi)始每周檢測(cè)模型組大鼠NFBG,觀察血糖升高水平。第8周末處死所有大鼠,采集血液標(biāo)本檢測(cè)FPG、NFBG、TC、TG、LDL、HDL、FINS及糖化血紅蛋白(Glycosylated hemoglobin A1c,HBA1c)等生化指標(biāo);留取腹主動(dòng)脈血管組織,HE染色法觀察血管組織形態(tài)學(xué)改變,q RT-PCR、Western Blot及免疫組化法分別檢測(cè)血管中HMGA1 m RNA和蛋白表達(dá)水平。利用Spearman法分別對(duì)FPG、NFBG、TC、TG、LDL、HDL和FINS與HMGA1 m RNA及蛋白表達(dá)水平進(jìn)行相關(guān)性分析。結(jié)果(1)起始時(shí)及喂養(yǎng)4周后,各組大鼠BW、FBG、NFBG、FINS及胰島素敏感指數(shù)(insulin sensitivity index,ISI)等指標(biāo)均無(wú)明顯差異(P0.05),起始時(shí)各組大鼠TG、TC、LDL及HDL水平亦無(wú)顯著差異(P0.05)。注射STZ后第3、10及21天測(cè)DM組大鼠NFBG均明顯升高(11.1mmol/l)。第8周末,DM組FBG、NFBG、Hb A1c、TG、TC、LDL及HDL水平較NC及HSFD組均明顯增加,而B(niǎo)W、FINS及ISI則明顯下降;HSFDs組的BW、FINS、TC及LDL水平較NC組明顯增加,ISI則明顯下降,以上差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。(2)熒光定量PCR結(jié)果顯示,DM組大鼠腹主動(dòng)脈HMGA1的m RNA表達(dá)水平明顯高于NC及HSFDs組(P0.05);HSFDs組HMGA1的m RNA表達(dá)水平稍高于NC組,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。(3)Western-blot及免疫組化結(jié)果均顯示,DM組大鼠腹主動(dòng)脈HMGA1蛋白表達(dá)水平明顯高于NC及HSFDs組(P0.05);HSFDs組HMGA1蛋白表達(dá)水平稍高于NC組,但差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);此外,免疫組化結(jié)果還顯示,HMGA1蛋白主要定位于血管平滑肌細(xì)胞(vascular smooth muscle cells,VSMCs)核中。(4)HE染色結(jié)果顯示,NC組血管內(nèi)膜平整、光滑,血管平滑肌細(xì)胞排列整齊、緊密;DM組血管內(nèi)膜欠光滑,血管中膜增厚,平滑肌細(xì)胞質(zhì)基質(zhì)疏松;HSFDs組血管形態(tài)介于前二者之間。對(duì)各組血管內(nèi)-中膜厚度(intimia-media thickness,IMT)進(jìn)行測(cè)量,結(jié)果顯示,HSFDs組血管內(nèi)-中膜厚度較NC組增加,DM組較NC及HSFDs組增加,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。(5)對(duì)FPG、NFBG、TC、TG、LDL、HDL及FINS與HMGA1 m RNA及蛋白表達(dá)水平進(jìn)行Spearman秩相關(guān)分析,結(jié)果顯示FBG、NFBG、TC及LDL水平均與大鼠主動(dòng)脈中HMGA1的表達(dá)水平呈正相關(guān)性(FBG:HMGA1m RNA:r=0.424,P=0.039,HMGA1 protein(IOD):r=0.549,P=0.005;NFBG:HMGA1 m RNA:r=0.584,P=0.003,HMGA1 protein(IOD):r=0.646,P=0.001;TC:HMGA1 m RNA:r=0.544,P=0.006,HMGA1protein(IOD):r=0.726,P=0.000;LDL:HMGA1 m RNA:r=0.636,P=0.001,HMGA1 protein(IOD):r=0.756,P=0.000)。結(jié)論(1)高糖高脂飼料喂養(yǎng)聯(lián)合小劑量鏈脲佐菌素腹腔注射可成功構(gòu)建大鼠2型糖尿病模型。(2)糖尿病大鼠主動(dòng)脈可發(fā)生中膜增厚,平滑肌細(xì)胞質(zhì)基質(zhì)疏松等形態(tài)改變;主動(dòng)脈組織中HMGA1m RNA及蛋白表達(dá)水平均增加。(3)FBG、NFBG、TC及LDL均與HMGA1表達(dá)水平呈正相關(guān)性,可能是影響動(dòng)脈血管組織中HMGA1表達(dá)變化的病理因素。
[Abstract]:Objective to construct rat model 2 diabetes by intraperitoneal injection of high glucose and high fat diet combined with Streptozotocin (STZ), and to observe the expression level of high mobility group protein A1 (high mobility group A1, HMGA1) m RNA and protein in the aortic vascular tissue of rats with diabetes, and to explore the possible influence on the change of the expression level of the protein. The pathological factors, try to provide new direction for the pathogenesis and prevention and control of diabetic vascular complications. Methods 4 weeks male SD rats were divided into normal control group (normol contol group, NC), high glucose and high fat feeding group (high-sucrose-high-fat diets group, HSFDs), diabetic model group (diabetes mellitus group,) DM) group.NC was fed with normal standard rat feed, group HSFDs and group DM were fed with high sugar and high fat diet. After 4 weeks of starting and feeding, the body weight (body weight, BW) was measured and fasting blood glucose (fasting blood glucose, FBG), non fasting blood glucose (nonfasting), glycerin, glycerin, and glycerol were three. Triglyceride, TG, low density lipoprotein (low density lipoprotein, LDL), high density lipoprotein (high density lipoprotein, HDL), and fasting insulin (fasting insulin) were injected into the abdominal cavity for the weekend. 3 days later, the rat model group was detected to observe the blood glucose increase. All rats were killed at the end of the eighth week, and blood samples were collected to detect the biochemical indexes such as FPG, NFBG, TC, TG, LDL, HDL, FINS and glycosylated hemoglobin (Glycosylated hemoglobin A1c, HBA1c), and to observe the vascular tissue of the abdominal aorta and observe the vascular histomorphology by HE staining. RNA and protein expression level. FPG, NFBG, TC, TG, LDL, HDL and FINS and HMGA1 m RNA and protein expression levels were analyzed by Spearman. Results (1) starting and feeding for 4 weeks, there were no significant differences between each group. There was no significant difference in TG, TC, LDL and HDL levels in each group (P0.05). The NFBG in DM group was significantly increased in 3,10 and 21 days after STZ injection (11.1mmol/l). At the end of the eighth week, DM group FBG. In addition, ISI decreased significantly, and the above differences were statistically significant (P0.05). (2) fluorescence quantitative PCR showed that the expression level of M RNA in abdominal aorta of DM rats was significantly higher than that in NC and HSFDs group (P0.05), but the level of HMGA1 in HSFDs group was slightly higher than that in the group, but there was no statistical significance (3) and immunohistochemical results. The expression of HMGA1 protein in abdominal aorta in DM rats was significantly higher than that in group NC and HSFDs (P0.05), and the expression level of HMGA1 protein in group HSFDs was slightly higher than that in NC group, but the difference was not statistically significant (P0.05). Moreover, the immunohistochemical results also showed that HMGA1 protein was mainly located in the nucleus of smooth muscle cells of the blood tube (vascular smooth). (4) The E staining showed that the vascular intima of the NC group was smooth and smooth and the vascular smooth muscle cells were arranged neatly and closely. The vascular intima in the DM group was not smooth, the membrane was thickened and the smooth muscle cytoplasm matrix was loose; the vascular morphology in the group HSFDs was between the first two. The blood vessel thickness (intimia-media thickness, IMT) was measured in each group, and the results showed HS The thickness of intravascular and middle membrane in group FDs was higher than that in group NC, and in group DM was higher than that in group NC and HSFDs, the difference was statistically significant (P0.05). (5) FPG, NFBG, TC, TG, LDL, HDL and expressions were correlated with the level of protein expression. FBG:HMGA1m RNA:r=0.424, P=0.039, HMGA1 protein (IOD): r=0.549, P=0.005, NFBG:HMGA1 m RNA:r=0.584. Intraperitoneal injection of small dose streptozotocin could successfully construct model 2 diabetic rats. (2) the aorta of the aorta could be thickened and the smooth muscle cytoplasm matrix was loose, and the expression level of HMGA1m RNA and protein in the aorta increased. (3) FBG, NFBG, TC and LDL were all positively correlated with the expression of HMGA1. Sex may be a pathological factor affecting the expression of HMGA1 in arterial tissue.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R587.2

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7 王志尊;鈣調(diào)神經(jīng)磷酸酶在大鼠主動(dòng)脈鈣化中的作用[D];河北醫(yī)科大學(xué);2008年

8 聶焱;慢性飲酒對(duì)大鼠主動(dòng)脈反應(yīng)性和血壓的影響及�；撬岬母深A(yù)作用[D];山西醫(yī)科大學(xué);2010年

9 鄭敬;鈣調(diào)神經(jīng)磷酸酶/活化T細(xì)胞核因子信號(hào)通路在大鼠主動(dòng)脈鈣化中的作用[D];河北醫(yī)科大學(xué);2009年

10 生玉平;大鼠主動(dòng)脈衰老進(jìn)程的比較蛋白質(zhì)組學(xué)研究[D];山東大學(xué);2013年

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