TLR4和TRAIL与糖尿病肾病的相关性研究

发布时间：2018-06-22 20:58

本文选题：TLR4 + TRAIL　；参考：《皖南医学院》2017年硕士论文

【摘要】：目的:通过检测TLR4和TRAIL在外周血单核细胞中表达水平的高低,同时检测血清中各种细胞因子的表达量,全面的分析TLR4和TRAIL与各细胞因子的相关性,结合多方面的结果,将有助于进一步探讨糖尿病肾病(DN)发病机制,有望成为DN早期诊断与愈后判断的指标,为DN的治疗提理论供依据。方法:收集2015年9月至2016年6月的糖尿病(DM)及DN患者各42例,并且全部符合相关诊断标准和排除标准,同时收集正常对照人群(CONTROL)42例;每个研究对象分别采血2管,提取血清、血浆,并分离抗凝管中的外周血单核细胞,使用qPCR的方法检测单核细胞中的TLR4mRNA及TRAILmRNA的含量;使用ELISA测定所收集血清中各类细胞因子的浓度;病例组与对照组之间患者的一般情况及所测量的生化指标采用方差分析或χ2检验;各组间细胞因子之间的浓度差异采用方差分析及Welch检验方法,因子间的相关性分析采用Pearson相关或Spearman's相关方法。结果:DN组、DM组、CONTROL组人群在性别、吸烟、饮酒与体质指数(BMI)方面的分布情况相近,未发现差异有统计学意义(P0.05),DN组患者在劳动强度方面低于CONTROL组研究对象,而在年龄上高于CONTROL组人群,DM组患者心理紧张的人数多于CONTROL组,CONTROL组初高中的人数多于DN组和DM组,差异有统计学意义(P0.05)。DN组、DM组和CONTROL组在总胆固醇(TC)、低密度脂蛋白(LDL-C)、总胆红素、直接胆红素、谷丙转氨酶(ALT)、尿素氮方面差异无统计学意义(P0.05);在空腹血糖(GLU)、收缩压、舒张压、甘油三酯(TG)、肌酐及尿酸方面,DN组的含量均高于CONTROL组,差异有统计学意义(P0.05)。在高密度脂蛋白(HDL-C)及谷草转氨酶(AST)方面,DN组的含量低于CONTROL组,差异有统计学意义(P0.05);DN组与DM组的焦虑、抑郁、睡眠质量评分要高于CONTROL组(P0.05);单因素Logistic回归分析显示年龄、焦虑、抑郁、睡眠、收缩压和舒张压是DN的危险因素,而劳动程度、文化程度及HDL-C是DN的保护因素(P0.05);多因素非条件Logistic回归显示睡眠质量与TG则是DN的危险因素,劳动强度与HDL-C是DN的保护因素(P0.05)。三组间TLR4mRNA、TRAILmRNA的表达量差异均有统计学意义(P0.01),DN组、DM组及CONTROL组在TLR4mRNA的表达量方面未发现差异(P0.05),DN组、DM组中TRAILmRNA的表达量要明显低于CONTROL组(P0.01);DN在IL-1、IL-6、TNF-α、MCP-1、NF-κB、hs-CRP方面表达量均高于CONTROL组,sTRAIL的浓度低于CONTROL组(P0.05),DN组中因子IL-1、TNF-a、MCP-1的浓度要高于DM组(P0.05)。分层分析显示三组间在≤50岁研究对象中,DN组中IL-1、IL-6、MCP-1的含量要高于CONTROL组(P0.05),sTRAIL、TRAILmRNA含量低于CONTROL组(P0.05),并且IL-6、MCP-1的表达测定量也高于DM组(P0.05);三组间在51-60岁研究对象中,DN组中IL-1、MCP-1的含量高于CONTROL组(P0.05),TRAILmRNA含量低于CONTROL组(P0.05),并且IL-1、MCP-1的含量高于DM组(P0.05);三组间在60岁研究对象中,DN组中IL-1、TNF-α、MCP-1的浓度均高于DM组、CONTROL组,而TRAILmRNA的表达量低于CONTROL组(P0.05)。Spearman's相关性分析可以看出TLR4mRNA与sTRAIL呈负相关(r=-0.188,P0.05),与其他因子则未发现相关性的存在(P0.05);TRAILmRNA则与IL-1、TNF-α、MCP-1、NF-κB、hs-CRP均呈负相关(P0.05),与sTRAIL的呈正相关(r=0.281,P0.05);对TLR4mRNA进行组间的相关性分析显示TLR4mRNA在DN组与DM组都与TRAILmRNA呈正相关(r=0.363,P0.05;r=0.540,P0.05)。TRAILmRNA在DN组中与NF-κB呈负相关(r=-0.422,P0.05);在≤50岁年龄段的研究对象中,TRAILmRNA与hs-CRP呈负相关(r=-0.346,P0.05),与sTRAIL呈正相关(r=0.394,P0.05);在60岁年龄段的研究对象中与IL-1呈负相关(r=-0.407,P0.05)。TLR4mRNA在DN组中且年龄60岁研究对象中与TNF-α呈负相关(r=-0.442,P0.05),与TRAILmRNA呈正相关(r=0.498,P0.05)。TRAILmRNA在DN组中且年龄为≤50岁的研究对象中显示与TNF-α呈正相关(r=0.749,P0.05),在DN组中≤50岁及51-60岁年龄组均与NF-κB呈负相关(r=-0.734,P0.05;r=-0.607,P0.05);在DN组中的年龄为≤50岁组中与hs-CRP呈负相关(r=-0.714,P0.05)。结论:在DN患者、DM患者及CONTROL人群中,DN患者血清中IL-1、IL-6、TNF-α、NF-κB、MCP-1、hs-CRP均高于CONTROL组,DN人群血清中sTRAIL的含量要低于DM组及CONTROL组;DN组及DM组外周血中TRAILmRNA的表达量均显著低于CONTROL组;TRAIL与TNF-α有着相似的途径,在体内的表达均受到年龄因素的影响,TNF-α在机体内表现为促炎因子,TRAIL在本研究中起抗炎作用,但在≤50岁的DN患者中与TNF-α存在高度的正相关;TRAILAmRNA的表达量与血清中L-1、IL-6、TNF-α、NF-κB、MCP-1、hs-CRP呈负相关。
[Abstract]:Objective: to detect the expression of TLR4 and TRAIL in peripheral blood mononuclear cells, to detect the expression of various cytokines in the serum, and to analyze the correlation between TLR4 and TRAIL and the various cytokines in a comprehensive way. Combined with many results, it will be helpful to further explore the pathogenesis of diabetic nephropathy (DN) and be expected to be the early diagnosis of DN. The index of judgment and judgment was based on the theory of DN. Methods: 42 cases of diabetes (DM) and DN patients from September 2015 to June 2016 were collected and all were in accordance with the relevant diagnostic criteria and exclusion criteria, and 42 cases of normal control group (CONTROL) were collected. 2 tubes were collected from each study, and serum, plasma, and isolation were extracted, respectively. The content of TLR4mRNA and TRAILmRNA in mononuclear cells was detected by qPCR in the peripheral blood mononuclear cells in the coagulant tube; the concentration of all kinds of cytokines in the collected serum was measured by ELISA; the general situation of the patients with the case group and the control group and the measured variance analysis of the biochemical indexes or the chi 2 test; the cytokine between each group The difference of concentration was analyzed by variance analysis and Welch test, and the correlation analysis between factors adopted Pearson related or Spearman's related methods. Results: the distribution of sex, smoking, drinking and body mass index (BMI) in group DN, DM and CONTROL group was similar, no statistical difference was found (P0.05), and the patients in group DN were in labor. The intensity is lower than the CONTROL group, and the age of group CONTROL is higher than that of group DM, and the number of patients in group DM is more than that of group CONTROL. The number of early and high school in group CONTROL is more than group DN and DM, the difference is statistically significant (P0.05).DN group, DM group and CONTROL group in total bile sterol (TC), low density lipoprotein, total bilirubin, direct There was no significant difference in bilirubin, ALT, and urea nitrogen (P0.05). In the fasting blood glucose (GLU), systolic pressure, diastolic pressure, triglyceride (TG), creatinine and uric acid, the content of DN group was higher than that of group CONTROL (P0.05). The content of DN group was low in high density lipoprotein (HDL-C) and glutamic pyruvic aminotransferase (AST). In group CONTROL, the difference was statistically significant (P0.05); the anxiety, depression, and sleep quality score of group DN and DM were higher than that of group CONTROL (P0.05); single factor Logistic regression analysis showed that age, anxiety, depression, sleep, systolic and diastolic pressure were the risk factors of DN, and the degree of labor, degree of education and HDL-C were the protective factors of DN (P0.05); multiple factors Non conditional Logistic regression showed that sleep quality and TG were risk factors for DN, labor intensity and HDL-C were the protective factors of DN (P0.05). There were significant differences in the expression of TLR4mRNA and TRAILmRNA between the three groups (P0.01). There was no difference in the expression of DN, DM and CONTROL groups. The amount of DN in IL-1, IL-6, TNF- alpha, MCP-1, NF- kappa B, hs-CRP is higher than that of the CONTROL group, and the concentration of sTRAIL is lower than that of the CONTROL group. The content of sTRAIL and TRAILmRNA was higher than that of group CONTROL (P0.05), and the content of TRAILmRNA was lower than that of CONTROL group (P0.05), and the expression of IL-6 and MCP-1 was higher than that of the DM group (P0.05). The content of IL-1 in the three group was higher than that in the group of DN, and the content was lower than that of the group. 5); among the three groups, the concentration of IL-1, TNF-, and MCP-1 in group DN was higher than that in group DM and CONTROL, while the expression of TRAILmRNA was lower than that of CONTROL group (P0.05).Spearman's. TNF- alpha, MCP-1, NF- kappa B, hs-CRP are negatively correlated (P0.05), and are positively correlated with sTRAIL (r=0.281, P0.05). In the study, TRAILmRNA was negatively correlated with hs-CRP (r=-0.346, P0.05), and was positively correlated with sTRAIL (r=0.394, P0.05). In the 60 year old age group, the negative correlation with IL-1 (r=-0.407, P0.05).TLR4mRNA was in the DN group and the 60 year old subjects were negatively correlated with the IL-1. AILmRNA in group DN and age younger than 50 years old showed positive correlation with TNF- alpha (r=0.749, P0.05). In group DN, there was a negative correlation with NF- kappa B (r=-0.734, P0.05; r=-0.607, P0.05) in the group of 50 and 51-60 years old. In the CONTROL population, the serum levels of IL-1, IL-6, TNF- a, NF- kappa B, MCP-1, and hs-CRP were all higher than those of the CONTROL group. The content of sTRAIL in the serum of DN population was lower than that in the group and group. The effect of TNF- alpha in the body is proinflammatory factor, and TRAIL plays an anti-inflammatory role in this study, but there is a positive correlation with TNF- alpha in DN patients less than 50 years old; the expression of TRAILAmRNA is negatively correlated with L-1, IL-6, TNF- a, NF- kappa B, MCP-1, hs-CRP in serum.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.2;R692.9

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