本文選題：糖尿病腎病 + 壞死性凋亡��； 參考：《遵義醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:構(gòu)建糖尿病腎病大鼠模型,探討壞死性凋亡信號(hào)途徑標(biāo)志物RIP1和RIP3在該病大鼠模型腎組織中的表達(dá)及意義。方法:雄性成年SD大鼠,標(biāo)準(zhǔn)化喂養(yǎng)。實(shí)驗(yàn)分為糖尿病腎病模型組(DN)和正常對(duì)照組(NC),每組2、4、8、12、16周5個(gè)觀察點(diǎn),每個(gè)時(shí)點(diǎn)6只大鼠,DN組采用鏈脲菌素(STZ)腹腔注射造模,50只SD大鼠注射STZ,2周時(shí)成模40只,未成模7只,死亡3只。處死前采取血液、尿液標(biāo)本,檢測(cè)血糖、血清肌酐、血清尿素和24小時(shí)尿蛋白;腎臟石蠟切片作蘇木素-伊紅(Hematoxylin-eosin,HE)和過(guò)碘酸-雪夫(Periodic Acid-Schiff,PAS)染色普通病理;運(yùn)用免疫組化、免疫熒光和蛋白印跡(Western Blot,WB)測(cè)定腎臟組織中受體交互作用蛋白1(Receptorinteraction protein1,RIP1)、受體交互作用蛋白3(Receptorinteraction protein 3,RIP3)及核轉(zhuǎn)錄因子Kappa B(Nuclearfactor-kappa B,NF-κB)的蛋白表達(dá),實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)(Real-Time polymerase chain reaction,RT-PCR)方法測(cè)定腎組織的RIP1、RIP3及NF-κBm RNA相對(duì)表達(dá)量。分析各指標(biāo)之間的相關(guān)性。結(jié)果:1.血、尿指標(biāo):1)血糖:(1)DN組不同時(shí)間點(diǎn)血糖比NC組明顯增加(P0.01)。(2)DN組血糖隨時(shí)間不斷增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05或P0.01)。2)24小時(shí)尿蛋白:(1)DN組不同時(shí)間點(diǎn)尿蛋白比NC組明顯增加(P0.01)。(2)DN組24小時(shí)尿蛋白定量隨時(shí)間不斷增加,差異有統(tǒng)計(jì)學(xué)意義(P0.01)。3)血清肌酐及尿素:(1)DN組不同時(shí)間點(diǎn)血清肌酐及尿素比NC組明顯增加(P0.01)。(2)DN組血清肌酐及尿素氮差異均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。2.腎臟病理:(1)DN組大鼠腎臟發(fā)白、腫大,可見(jiàn)不同程度的腎小球肥大、細(xì)胞數(shù)增多、系膜基質(zhì)增多、部分腎小管擴(kuò)張及空泡樣變性。(2)DN組不同時(shí)間點(diǎn)腎重指數(shù)比NC組明顯增加(P0.01)。(3)DN組腎重指數(shù)隨時(shí)間不斷增加,差異有統(tǒng)計(jì)學(xué)意義(P0.01或P0.05)。3.免疫組化及熒光:(1)RIP1、RIP3及NF-κB主要表達(dá)于腎小球中;(2)DN組RIP1、RIP3及NF-κB不同時(shí)間點(diǎn)比NC組均明顯增加(P0.01)。(3)DN組以上三個(gè)指標(biāo)表達(dá)量不同時(shí)間點(diǎn)比較,均為2W與4W相比差異無(wú)統(tǒng)計(jì)學(xué)意義,8W、12W、16W顯著高2W、4W,差異有統(tǒng)計(jì)學(xué)意義(P0.01),8W與12W相比差異無(wú)統(tǒng)計(jì)學(xué)意義,16W顯著高于8W、12W,差異有統(tǒng)計(jì)學(xué)意義(P0.01或P0.05)。(4)DN組NF-κB的蛋白表達(dá)量與RIP1、RIP3呈正相關(guān)性;RIP1、RIP3與尿蛋白、腎重指數(shù)、血清肌酐及尿素均呈正相關(guān)。4.蛋白印跡:(1)DN組不同時(shí)間點(diǎn)RIP1、RIP3及NF-κB蛋白表達(dá)均高于NC組(P0.01)。(2)DN組以上三個(gè)指標(biāo)表達(dá)量不同時(shí)間點(diǎn)比較,均為2W與4W相比差異無(wú)統(tǒng)計(jì)學(xué)意義,8W、12W、16W顯著高于2W、4W,差異有統(tǒng)計(jì)學(xué)意義(P0.01),12W與8W相比差異無(wú)統(tǒng)計(jì)學(xué)意義,16W顯著高于8W和12W,差異有統(tǒng)計(jì)學(xué)意義(P0.01)。(3)DN組NF-κB與RIP1、RIP3呈正相關(guān)性;RIP1、RIP3與尿蛋白、腎重指數(shù)、肌酐和尿素均呈正相關(guān)。5.Real-Time PCR:(1)DN組不同時(shí)間點(diǎn)RIP1、RIP3及NF-κB m RNA表達(dá)均高于NC組(P0.01),差異有統(tǒng)計(jì)學(xué)意義(P0.01)。(2)DN組以上三個(gè)指標(biāo)表達(dá)量不同時(shí)間點(diǎn)比較,均為2W與4W差異無(wú)統(tǒng)計(jì)學(xué)意義,8W、12W、16W顯著高于2W、4W,差異有統(tǒng)計(jì)學(xué)意義(P0.01),12W與8W相比差異無(wú)統(tǒng)計(jì)學(xué)意義,16W顯著高于8W和12W,差異有統(tǒng)計(jì)學(xué)意義(P0.01)。(3)DN組NF-κB與RIP1、RIP3呈正相關(guān)性;RIP1、RIP3與尿蛋白、腎重指數(shù)均呈正相關(guān),RIP3與肌酐和尿素呈正相關(guān),RIP1與肌酐和尿素?zé)o明顯相關(guān)性。結(jié)論:1.在糖尿病腎腎病大鼠腎組織中,壞死性途徑凋亡可能被激活,可能與糖尿病腎病大鼠腎組織炎癥反應(yīng)和損傷有關(guān)。2.NF-κB和壞死性凋亡途徑相互作用可能促進(jìn)炎癥反應(yīng)和糖尿病腎病大鼠的腎組織損傷。
[Abstract]:Objective: to construct a diabetic nephropathy rat model and explore the expression and significance of RIP1 and RIP3 in the renal tissue of the rat model of necrotic apoptosis signal. Methods: male adult SD rats, standardized feeding. The experimental group was divided into diabetic nephropathy model group (DN) and normal group (NC), each group of 2,4,8,12,16 week 5 observation points, each time point. In 6 rats, group DN was intraperitoneally injected with streptozotocin (STZ), 50 SD rats were injected with STZ, 40 in 2 weeks, 7 without model, and 3 dead. Before death, blood, urine specimens, blood glucose, serum creatinine, serum urea and 24 hours proteinuria were detected, and renal paraffin section was made of Hematoxylin-eosin, HE and periodate - Schiff (P). Eriodic Acid-Schiff, PAS) staining common pathology, using immunohistochemistry, immunofluorescence and Western Blot (WB) for the determination of receptor interaction protein 1 (Receptorinteraction protein1, RIP1) in renal tissue, receptor interaction protein 3 (Receptorinteraction protein 3, RIP3) and nuclear transcription factors. The protein expression of - kappa B, the real-time fluorescent quantitative polymerase chain reaction (Real-Time polymerase chain reaction, RT-PCR) method for determining the relative expression of RIP1, RIP3 and NF- kappa Bm in renal tissue. Analysis of the correlation between the indexes. Results: 1. blood, urine index: 1) blood glucose: (1) blood glucose at different time points in the DN group was significantly higher than that of the group. (2) blood group blood The sugar was increased with time, the difference was statistically significant (P0.05 or P0.01).2) 24 hour urine protein: (1) the urine protein in the DN group was significantly increased at different time points (P0.01). (2) the 24 hour urine protein in the DN group increased with time, the difference was statistically significant (P0.01).3) serum creatinine and urea: (1) the serum creatinine and urea at different time points in the DN group Compared with group NC (P0.01), (2) there was no significant difference in serum creatinine and urea nitrogen in group DN (P0.05).2. renal pathology: (1) kidney white and swelling in group DN, the number of glomerular cells increased, the number of cells increased, the mesangial matrix increased, partial renal tubule dilation and vacuolating degeneration. (2) the renal weight index of group DN at different time points was compared with that of the NC group (3) (3) the renal weight index of group DN increased with time, the difference was statistically significant (P0.01 or P0.05).3. immunization and fluorescence: (1) RIP1, RIP3 and NF- kappa B were mainly expressed in the glomeruli; (2) DN RIP1, RIP3 and different time points were significantly increased. (3) the expression of three indicators at different time points above The difference between 2W and 4W was not statistically significant, 8W, 12W, 16W were significantly higher 2W, 4W, and the difference was statistically significant (P0.01). The difference between 8W and 12W was not statistically significant. (4) the difference was statistically significant. The index, serum creatinine and urea were positively correlated with.4. blot: (1) the expression of RIP1, RIP3 and NF- kappa B protein at different time points in group DN were higher than that of NC group (P0.01). (2) the differences between 2W and 4W were not statistically significant compared with those of 2W and 4W. There was no statistical significance compared with 8W, 16W was significantly higher than 8W and 12W (P0.01). (3) there was a positive correlation between NF- kappa B and RIP1, RIP3 in DN group; RIP1, RIP3 and urine protein, renal weight index, creatinine and urea were positively correlated (1) There were statistical significance (P0.01). (2) the difference between 2W and 4W was not statistically significant, 8W, 12W, 16W were significantly higher than 2W, 4W, and the difference was statistically significant (P0.01). There was no statistical significance in 12W and 8W. (3) the difference was statistically significant. IP3 has positive correlation; RIP1, RIP3 and urine protein, renal weight index are positively correlated, RIP3 is positively correlated with creatinine and urea, RIP1 has no significant correlation with creatinine and urea. Conclusion: 1. in diabetic renal nephrotic rats, the apoptosis of necrotic pathway may be activated, which may be related to the inflammatory reaction and injury of renal tissue in diabetic nephropathy rats, which may be related to.2 The interaction between.NF- B and necrotic apoptotic pathway may promote inflammatory reaction and renal tissue damage in diabetic nephropathy rats.

【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R587.2;R692.9

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2 趙陸斌;六味地黃丸對(duì)糖尿病腎病大鼠MCP-1、NF-kB表達(dá)的影響及保護(hù)機(jī)制的研究[D];浙江中醫(yī)藥大學(xué);2015年

3 倪偉建;小檗堿對(duì)糖尿病腎病大鼠腎臟保護(hù)作用及對(duì)腎小球系膜細(xì)胞前列腺素EP1受體信號(hào)通路的調(diào)控[D];安徽醫(yī)科大學(xué);2015年

4 王春寧;活血化瘀通絡(luò)中藥對(duì)糖尿病腎病大鼠的干預(yù)作用及對(duì)AT_2R的影響[D];河北醫(yī)科大學(xué);2015年

5 車(chē)瑩;DPP-Ⅳ抑制劑西格列汀對(duì)糖尿病腎病大鼠ERK1/2信號(hào)通路的影響[D];山西醫(yī)科大學(xué);2015年

6 閆婧;西格列汀對(duì)糖尿病腎病大鼠腎臟JNK信號(hào)通路的影響[D];山西醫(yī)科大學(xué);2015年

7 周雪娜;益腎湯對(duì)糖尿病腎病大鼠炎癥因子IL-6、TNF-α的影響[D];山東中醫(yī)藥大學(xué);2015年

8 陳磊;芪術(shù)膠囊干預(yù)糖尿病腎病大鼠的代謝組學(xué)研究[D];山西中醫(yī)學(xué)院;2015年

9 劉明;藕節(jié)對(duì)糖尿病腎病大鼠腎組織p-JAK2、p-STAT3及VEGF表達(dá)的影響[D];山西醫(yī)科大學(xué);2014年

10 劉曉敏;雙重阻斷RAS系統(tǒng)對(duì)實(shí)驗(yàn)性糖尿病腎病大鼠腎組織Sonic hedgehog信號(hào)通路激活的影響[D];桂林醫(yī)學(xué)院;2016年

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