高糖条件下IGF-1对大鼠胃平滑肌细胞内质网应激与自噬的影响及机制

发布时间：2020-12-31 21:05

　　研究背景:糖尿病胃轻瘫（Diabetic gastroparesis,DGP）是糖尿病胃肠道最常见的并发症,常表现为非机械性梗阻情况下的胃动力障碍及胃排空延迟,目前DGP发病机制尚不明确。截止目前,自主神经病变、急性血糖升高及间质卡哈尔细胞（Interstitial Cahal cell,ICC）的损伤和缺失被认为是影响胃动力的主要因素。胰岛素样生长因子-1（Insulin-like growth factor-1,IGF-1）主要由肝脏分泌,是结构与胰岛素有一定相似性的蛋白多肽。近年来的众多研究发现,IGF-1与糖尿病慢性并发症的发生、发展有着密切的关系。磷脂酰肌醇3激酶（Phosphoinositide3-kinase,PI3K）-丝氨酸蛋白激酶（Akt）通路是细胞内重要的信号通路。PI3K-Akt通路在许多糖尿病并发症中被激活,且为IGF-1下游信号。Zhang等人研究发现,PI3K-Akt通路参与了 DGP发生,并且随胃轻瘫的发展其表达逐渐降低。蛋白激酶C（Protein kinase C,PKC）家族有3种,包括经典型、新型及非典型PKC,其中经典型分为PKCα、PKCβⅠ、... 

【文章来源】：延边大学吉林省 211工程院校

【文章页数】：92 页

【学位级别】：博士

【文章目录】：
摘要
ABSTRACT
ABBEVIATIONS
1. INTRODUCTION
    1.1 The pathogenesis of DGP
        1.1.1 Interstitial cells of Cajal （ICC）
        1.1.2 Extrinsic nervous system
        1.1.3 Enteric nervous system lesion
        1.1.4 Gastric smooth muscle cells
        1.1.5 Immune Cells
        1.1.6 Macrophages
        1.1.7 Hyperglycemia
    1.2 Insulin-like growth factor-1 and diabetes
    1.3 Endoplasmic reticulum stress and autophagy
    1.4 Study design
2. MATERIALS AND METHODS
    2.1 Materials
    2.2 Methods
        2.2.1 Diabetic rat model establishment and grouping
        2.2.2 Immunofluorescence staining of tissue sections
        2.2.3 Western blot analysis of GRP78,CHOP, LC3Ⅱ, PI3K, p-Akt, PKCα, PKCβ1 proteinexpression in rat gastric smooth muscle tissues
        2.2.4 Primary culture of rat gastric smooth muscle cells and grouping
        2.2.5 Detection of PKC activity in rat gastric smooth muscle cells by ELISA method
2+ concentration in rat gastric smoothmuscle cells by confocal laser-scanning microscopy">        2.2.6 Detection of canges in intracellular Ca²⁺ concentration in rat gastric smoothmuscle cells by confocal laser-scanning microscopy
        2.2.7 Detection of GRP78 and LC3 in rat gastric smooth muscle cells by confocallaser-scanning microscopy
        2.2.8 Western blot analysis of GRP78,CHOP, LC3Ⅱ,PI3K, p-Akt, PKCα, PKCβ1,p-PKCα, p-PKCβ1 protein expression in gastric smooth muscle cells
        2.2.9 Statistical analysis
3. RESULTS
    3.1 Results of immunofluorescence staining of GRP78 and LC3 in rat gastric smoothmuscle tissues
    3.2 Western blot assay results of GRP78,CHOP, LC3Ⅱ,PI3K, p-Akt, PKCα,PKCβ1protein expression in rat gastric smooth muscle tissues of each group
    3.3 Identification of gastric smooth muscle cell
    3.4 PKC activity in rat gastric smooth muscle cells as detected by ELISA method
2+ concentration in rat gastric smooth muscle cells as detected byconfocal laser-scanning microscopy">    3.5 Changes in Ca²⁺ concentration in rat gastric smooth muscle cells as detected byconfocal laser-scanning microscopy
    3.6 GRP78, LC3 expression in rat gastric smooth muscle cells as detected by confocallaser-scanning microscopy
    3.7 Expression of GRP78, CHOP, LC3Ⅱ,PI3K,p-Akt, PKCα, PKCβ1,p-PKCα, p-PKCβ1protein expression in vitro-cultured rat gastric smooth muscle cells as detected by westernblot assay
4. DISCUSSION
5. CONCLUSIONS
REFERENCES
致谢
攻读博士期间发表的论文


【参考文献】：
期刊论文
[1]Sequential blood purification therapy for critical patients with hyperlipidemic severe acute pancreatitis[J]. Hong-Liang Wang,Kai-Jiang Yu.  World Journal of Gastroenterology. 2015(20)
[2]C-type natriuretic-peptide-potentiated relaxation response of gastric smooth muscle in streptozotocin-induced diabetic rats[J]. Ying-Lan Cai, Dong-Yuan Xu, Xiang-Lan Li, Zheng Jin, Department of Physiology, Yanbian University School of Medicine, Yanji 133000, Jilin Province, China Zhang-Xun Qiu, Wen-Xie Xu, Department of Physiology, Shanghai Jiaotong University School of Medicine, Shanghai 200240, China.  World Journal of Gastroenterology. 2009(17)



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