慢性间歇性低氧激活内质网应激介导大鼠肾脏损伤及复氧后肾脏功能恢复的机制研究

发布时间：2018-06-22 03:42

本文选题：阻塞性睡眠呼吸暂停低通气综合征 + 慢性间歇性低氧　；参考：《兰州大学》2017年硕士论文

【摘要】：目的探讨阻塞性睡眠呼吸暂停低通气综合征(OSAHS)在普通饮食、高脂饮食条件下引起肾脏损伤的机制以及复氧后肾脏组织结构以及肾脏功能的恢复。本研究是以慢性间歇性低氧(CIH)、高脂饮食模拟OSHAS及高脂饮食,其通过激活内质网应激反应(ERS)介导大鼠肾脏的损伤;并以复氧模拟临床持续气道内正压通气(CPAP)治疗,观察肾脏组织结构及功能的恢复。为临床的治疗提供新的诊疗思路。方法将48只SPF级健康的雄性Wistar大鼠,随机分为4组,每组12只:A组:对照组(常氧及普通饮食组)、B组:高脂组(常氧及高脂饮食组)、C组:CIH组(CIH及普通饮食组)、D组:CIH合并高脂饮食组(CIH及高脂饮食组)。6周后每组随机选取6只,检测血清中胱抑素C(Cys-C)、高密度脂蛋白(HDL)、并行肾脏的CHOP蛋白免疫组化,并苏木精-伊红(HE)染色后在显微镜下观察肾脏病理结构变化,以及电镜下观察肾小球及肾小管超微结构变化。再在原饮食条件下复氧2周,再次重复上述检测,并进行比较。结果B组、C组、D组中肾早损指标Cys-C水平较A组显著升高,且在D组中值最高(均P0.05);B组、D组中血脂指标HDL水平较A组显著下降(均P0.05),C组与A组比较有下降,但无显著差异(P0.05);B组、C组、D组中ERS标志性蛋白CHOP平均光密度值与A组比较显著升高,且在D组中值最高(均P0.05)。HE染色光镜下观察发现:A组肾脏组织未见明显异常,在B组、C组中肾小球未见明显异常,肾小管上皮细胞小部分水肿,D肾小球未见明显异常,组肾小管上皮细胞大部分水肿;电镜下观察,B组、C组近曲肾小管上皮细胞局灶性核固缩,刷状缘稀疏并脱落,D组中有大片肾小管上皮细胞核固缩,刷状缘稀疏并脱落。并复氧2周后Cys-C水平在A组、B组与复氧前比较无显著变化(均P0.05),在C组、D组中与复氧前比较显著降低(均P0.05);复氧后HDL水平在A组、B组与复氧前比较有下降,但无显著性差异(均P0.05),在C组与复氧前比较显著升高(P0.05),在D组有升高,但无显著性差异(P0.05)。复氧后CHOP蛋白平均光密度水平在肾脏中的表达,A组、B组水平与复氧前无明显差异(均P0.05),C组、D组水平与复氧前比较明显降低(均P0.05)。光镜及电镜下观察:A组未见明显异常,B组与复氧前比较水肿稍有加重,复氧前C组、D组与复氧后比较肾脏组织改善。结论1.CIH合并高脂饮食的模型建立。2.CIH、高脂可通过激活ERS介导近曲肾小管上皮的凋亡,使肾脏的超微结构发生改变,进而肾脏功能损伤。3.复氧可通过改善ERS,使肾脏结构与肾功能改善。
[Abstract]:Objective to investigate the mechanism of renal injury induced by obstructive sleep apnea hypopnea syndrome (OSAHS) under general diet and high fat diet, and to investigate the recovery of renal structure and function after reoxygenation. In this study, chronic intermittent hypoxia (CIH), high-fat diet, OSHAS and high-fat diet were used to induce renal injury in rats by activating endoplasmic reticulum stress response (ERS), and reoxygenation was used to simulate clinical continuous positive airway pressure ventilation (CPAP). To observe the recovery of renal tissue structure and function. To provide a new way of diagnosis and treatment for clinical treatment. Methods 48 SPF healthy male Wistar rats were randomly divided into 4 groups. 12 rats in each group: control group (normoxic group and normal diet group) group B: high fat group (normoxic and high fat diet group) and C group (CIH and general diet group), group D: CIH combined with high-fat diet group (CIH and high-fat diet group) 6 weeks after 6 weeks, 6 rats in each group were randomly selected. Cystatin C (Cys-C), high density lipoprotein (HDL), chop protein immunohistochemical staining were used to detect cystatin C (Cys-C) and high density lipoprotein (HDL) in serum. The pathological changes of kidney were observed under microscope after hematoxylin-eosin (HE) staining, and ultrastructural changes of glomeruli and renal tubules were observed under electron microscope. After reoxygenation for 2 weeks, the above tests were repeated and compared. Results the level of Cys-C in group B was significantly higher than that in group A, and the level of HDL in group D was significantly lower than that in group A (P0.05), and the level of HDL in group D was significantly lower than that in group A (P0.05). But there was no significant difference (P0.05). The mean optical density of ERS marker protein chop in group C was significantly higher than that in group A (P0.05). There was no obvious abnormality in glomeruli in group B and C, and there was no obvious abnormality in small portion of edema of renal tubular epithelial cells and no obvious abnormality in D glomeruli, and most of renal tubular epithelial cells in group B were oedema. The focal nuclear pyknosis of proximal tubule epithelial cells in group B and C was observed under electron microscope. In group D, a large number of renal tubular epithelial nuclei were condensed and abscission. After reoxygenation, the level of Cys-C did not change significantly in group A and group B before reoxygenation (P0.05), but decreased significantly in group C and group D (P0.05), and the level of HDL decreased after reoxygenation in group A and group B before reoxygenation. But there was no significant difference (P0.05), in group C and before reoxygenation significantly increased (P0.05), in group D increased, but there was no significant difference (P0.05). Expression of chop protein in kidney after reoxygenation there was no significant difference between group A and group B (P0.05). Under light and electron microscope, there was no obvious abnormal edema in group B compared with that before reoxygenation, and the renal tissue was improved in group C before reoxygenation and in group D after reoxygenation. Conclusion 1. CIH combined with high fat diet can induce apoptosis of proximal tubule epithelium through activation of ERS, which can change the ultrastructure of kidney and then damage renal function. 2. CIH can induce apoptosis of proximal tubule epithelium by activation of ERS. Reoxygenation can improve renal structure and function by improving ERS.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R766

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