本文選題：apelin　切入點：1型糖尿病　出處：《華中科技大學》2014年博士論文

【摘要】：糖尿病是由胰島素的分泌障礙和／或胰島素的作用障礙所引起的,以長期高血糖為特征的代謝性疾病。糖尿病的長期慢性高血糖會引起器官的功能障礙和衰竭,造成各種糖尿病并發(fā)癥的發(fā)生。糖尿病腎病是一種常見糖尿病微血管并發(fā)癥,以腎小球硬化和腎病綜合征為特征,也被認為是導致終末期腎衰竭的主要原因。肥胖型糖尿病腎病的病發(fā)過程中常伴隨著腎臟缺血再灌注損傷(ischemia/reperfusion injury, I/R injury)的發(fā)生,但對于糖尿病及糖尿病腎病和腎臟缺血再灌注損傷的分子機制及其防治手段尚需進一步研究。Apelin是一類新型脂肪因子,具有調(diào)節(jié)血壓、胰島素分泌和能量代謝等多種生物功能,其中apelin-13是apelin家族中最豐富且活性最高的成員。本論文研究了apelin-13是否能有效地抑制糖尿病所致的胰臟和腎臟病變,是否能抑制缺血/再灌注引起的腎臟損傷。 本論文以1型糖尿病模型Akita小鼠為實驗對象,研究1型糖尿病導致的胰臟和腎臟病變；并以高糖刺激的HBZY-1和MES13細胞作為模擬糖尿病腎病的體外細胞模型。我們還以單側(cè)腎結(jié)扎的wistar大鼠作為腎臟缺血再灌注損傷的動物模型；以在低氧(1%02)條件下進行無糖無血清培養(yǎng)基培養(yǎng)的HBZY-1和NRK-52E細胞作為模擬腎臟缺血再灌注損傷的體外模型。本論文通過蛋白質(zhì)免疫印跡、免疫組織化學染色、免疫熒光染色、聚合酶鏈式反應和染色質(zhì)免疫共沉淀等手段,從分子水平、細胞水平和病理水平研究了apelin對胰臟和腎臟損傷的治療作用的分子機理。 我們首先對Akita小鼠進行為期兩個月的apelin-13的尾靜脈注射,結(jié)果顯示apelin-13治療明顯改善糖尿病引起的胰臟中胰島面積和胰島素含量減少。進一步的研究表明,apelin-13能抑制糖尿病誘導內(nèi)質(zhì)網(wǎng)應激反應(endoplasmic reticulum stress, ER stress)中PERK和IREla通路的激活,降低分子伴侶(GRP78, calnexin和HSP70)在Akita小鼠胰臟中的表達水平,并激活糖尿病所抑制的AKT、AMPK和ERK通路。其次,在Akita小鼠中, apelin-13不僅降低了糖尿病引起的總尿量、尿總蛋白和腎小球濾過率等腎功能相關(guān)生理指標的上升,還抑制了糖尿病導致的腎小球肥大、系膜擴張和腎臟炎癥。本論文首次發(fā)現(xiàn)apelin-13明顯的抑制了由糖尿病或高糖或丁酸鈉引起的NF-κB和組蛋白的乙�；芳せ顚е碌难装Y因子的表達。并且apelin-13可以上調(diào)糖尿病或高糖或丁酸鈉抑制的HDAC1表達。另外,在缺血再灌注引起腎損傷的大鼠中,損傷前腹腔注射apelin-13能顯著降低腎臟缺血再灌注損傷引起的腎小管壞死,細胞凋亡和組蛋白的高甲基化水平。在HBZY-1和NRK-52E細胞進行的缺氧實驗顯示：apelin-13通過抑制Tgf-β和組蛋白的高甲基化水平引起的凋亡反應,達到對細胞的保護作用。此外,在HBZY-1細胞中過表達apelin也能抑制I/R損傷引起的Tgf-P和組蛋白的高甲基化水平,有效抑制I/R損傷引起的細胞凋亡。 本論文研究結(jié)果顯示,在Akita小鼠中,apelin-13通過抑制糖尿病引起的內(nèi)質(zhì)網(wǎng)應激反應并促進胰臟中胰島素的表達。Apelin-13能降低糖尿病引起的腎臟中組蛋白的高乙�；�,抑制NF-κB和MAPK通路的激活,進而抑制腎臟的炎癥反應。此外,apelin-13還可抑制缺血再灌注損傷所致的Tgf-β通路和組蛋白的高甲基化水平,最終抑制細胞凋亡。Apelin-13不但對1型糖尿病及糖尿病腎病有良好治療效果,對急性腎損傷也有顯著療效。本研究不僅為探索1型糖尿病及糖尿病腎病、急性腎損傷的分子機制提供指導意見,并且為1型糖尿病及糖尿病腎病、急性腎損傷的藥物治療提供了新的研究方向。
[Abstract]:Diabetes is caused by insulin secretion function disorder disorder and / or insulin, metabolic disease characterized by long-term high glucose. Diabetes chronic hyperglycemia can cause organ dysfunction and failure, caused by various complications of diabetes. Diabetic nephropathy is a common microvascular complication of diabetes, glomerular sclerosis and nephrotic syndrome characteristics, is also considered to be the leading cause of end-stage renal failure. Renal obese diabetic disease process often accompanied by renal ischemia reperfusion injury (ischemia/ reperfusion injury, I/R injury) occurred, but the molecular mechanism for diabetes and diabetic nephropathy and renal ischemia reperfusion injury and the treatment needs further study.Apelin is a kind of new fat factor, can regulate the blood pressure, insulin secretion and energy metabolism etc. Apelin-13 is the most abundant and highly active member of Apelin family. In this paper, we studied whether apelin-13 can effectively inhibit pancreatic and renal lesions caused by diabetes and whether it can inhibit renal injury induced by ischemia / reperfusion.
In this paper, a model of type 1 diabetes Akita mice as experimental subjects, type 1 diabetes mellitus in pancreas and kidney disease; and glucose stimulated HBZY-1 and MES13 cells as in vitro cell model of diabetic nephropathy. We also used Wistar rats as the animal model of unilateral renal ligation of renal ischemia reperfusion injury; to in hypoxia (1%02) in vitro model of sugar free cultured HBZY-1 and NRK-52E cells as a model of renal ischemia reperfusion injury conditions. This paper through Western blot, immunohistochemistry, immunofluorescence staining, polymerase chain reaction and chromatin immunoprecipitation method, from the molecular level, molecular study on the mechanism of the therapeutic effect of apelin on the pancreas and kidney damage in the cell level and pathological level.
We first intravenous injection for two months apelin-13 to Akita mice, showed decreased islet area and insulin content of apelin-13 treatment significantly improved diabetic pancreas. Further studies showed that apelin-13 can inhibit the diabetes induced endoplasmic reticulum stress response (endoplasmic reticulum stress, ER stress and IREla PERK) in the activation pathway the lower molecular chaperones (GRP78, calnexin and HSP70) on the expression level of Akita mice in the pancreas, and activate diabetes inhibition of AKT, AMPK and ERK pathways. Secondly, in Akita mice, apelin-13 not only reduces the total amount of urine caused by diabetes, the rise of related physiological indexes of total urinary protein and glomerular filtration rate. Renal function, also inhibits diabetes induced glomerular hypertrophy, mesangial expansion and inflammation of the kidneys. This is the first time that apelin-13 significantly suppressed by diabetes. The expression of acetylation pathway disease or high glucose or sodium butyrate induced NF- kappa B and histone activation of inflammatory factors caused. And apelin-13 can up regulate the expression of diabetes or high glucose or sodium butyrate inhibited HDAC1. In addition, in ischemia reperfusion induced renal injury in rats, injury of anterior abdominal cavity injection can significantly reduce the apelin-13 renal ischemia reperfusion injury of renal tubular necrosis caused by high levels of methylation, apoptosis and histone. Hypoxia experiments were carried out in HBZY-1 and NRK-52E cells: the apoptosis reaction induced by apelin-13 through inhibition of Tgf- methylation level and protein beta group, achieve protective effect on cells. In addition, the high level of methylation Tgf-P and histone Apelin overexpression in HBZY-1 cells can inhibit I/R damage induced apoptosis, inhibit I/R damage caused.
The results of this study demonstrate that in Akita mice, apelin-13 via endoplasmic reticulum stress inhibition caused by diabetes and high levels of acetylation promotes the expression of.Apelin-13 in pancreatic insulin can reduce diabetes induced renal histone, inhibited the activation of NF- K B and MAPK pathway, and then inhibit the inflammatory reaction of the kidney. In addition, apelin-13 can inhibit the ischemia and reperfusion injury caused by high levels of methylation and histone Tgf- beta pathway, ultimately inhibit apoptosis of.Apelin-13 not only has a good effect in treating type 1 diabetes mellitus and diabetic nephropathy, also has significant curative effect on acute renal injury. This study not only for type 1 diabetes mellitus and diabetic nephropathy, provide guidance molecules the mechanism of acute renal injury, and type 1 diabetes mellitus and diabetic nephropathy, which provides a new research direction in the treatment of acute kidney injury.

【學位授予單位】：華中科技大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R587.2;R692

【參考文獻】

相關(guān)期刊論文 前1條

1 ;Apelin and vascular endothelial growth factor are associated with mobilization of endothelial progenitor cells after acute myocardial infarction[J];Journal of Biomedical Research;2012年06期

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本文編號：1699449