基于中医证候的糖尿病微血管病变危险因素分析及糖尿病视网膜病变易感基因初探

发布时间：2018-05-03 23:36

本文选题：糖尿病微血管病变 + 中医证候　；参考：《中国中医科学院》2017年硕士论文

【摘要】：目的:通过分析糖尿病患者一般资料、临床症状、中医证候及实验室检查,找到与糖尿病微血管病变相关的危险因素,在此基础上,通过配对糖尿病视网膜病变患者的危险因素后,筛查患者独立的易感基因,为今后2型糖尿病微血管病变的防治及糖尿病视网膜病变的深入研究提供依据。方法:(1)采用自行编制的调查表,收集北京、石家庄、保定等13家医院在2016年3月至2017年1月期间内分泌门诊和住院部T2DM患者的相关信息,进行横断面分析。采用Logistic回归分析对各因素进行单因素及多因素分析。(2)在前期找出DR危险因素的基础上,将具备DR所有危险因素的DR患者与未并发DR的T2DM患者的相关信息(包括一般资料,临床症状及实验室检查)进行匹配,选取相关信息完全一致的DR和非DR患者为一对,纳入研究,提取患者静脉全血的DNA,采用ILLUMINA人类SNP分型检测芯片技术,对DR患者242901个SNP位点进行筛查,找出所有纳入组均存在差异的位点,通过NCBI数据库(http://www.ncbi.nlm.nih.gov/)、UCSC 数据库(http://genome.ucsc.edu/)、lncR NASNP(http://bioinfo.life.hust.edu.cn/)数据库和 HGNC 数据库(http://www.gen enames.org/)查询差异位点所处的基因,再通过KEGG数据库(http://www.kegg.jp/)查找基因代谢通路。结果:最终有2041例糖尿病患者进入调查研究,其中有微血管病变的患者1318例(并发糖尿病视网膜病变患者734例,仅并发糖尿病视网膜病变患者138例;并发糖尿病肾病患者468例,仅并发糖尿病肾病患者的96例;并发糖尿病周围神经病变患者1044例,仅并发糖尿病周围神经病变患者379例),未发生微血管病变的患者723例。单因素logistic回归分析结果显示年龄(OR=1.016,95%CI 1.008-1.023,P=0.000),病程(OR=1.071,95%CI 1.005-1.086,P=0.000),家族史(OR=1.306,95%CI 1.085-1.573,P=0.005),高血压病(OR=1.651,95%CI 1.376-1.983,P=0.000),高脂血症病(OR=1.270,95%CI 1.056-1.529,P=0.011),心血管病(OR=1.978,95%CI 1。608-2.433,P=0.000),脑血管病(OR=1.269,95%CI 1.009-1.595,P=0.042),BMI(OR=1.035,95%CI 1.011-1.059,P=0.003),FBG(OR= 1.025,95%CI 1.001-1.050,P=0.045),2hPBG(OR=1.112,95%CI 1.083-1.142,P=0.000),HbA1c(OR=1.100,95%CI 1.053-1,150,P=0.000),SBP(OR=1.008,95%CI 1.002-1.013,P=0.004),DBP(OR=0.997,95%CI 0.995-1.000,P=0.024),瘀血阻滞证(OR=1.247,95%CI 1.038-1.502,P=0.018),痰瘀互阻证(OR=1.460,95%CI 1.144-1.865,P=0.002),阴虚燥热证(OR=0.444,95%CI 0.326-0.606,P=0.000),脾肾阳虚证(O R=2.335,95%CI 1.549-3.520,P=0.000)与糖尿病微血管病变有相关性。经多因素回归分析后发现病程(OR=1.052,95%CI 1.036-1.069,P=0.000),家族史(O R=1.241,95%CI 1.016-1.517,P=0.035),高血压病(OR=1.315,95%CI 1.052-1.643,P=0.035),心血管病(OR=1.522,95%CI 1.204-1.924,P=0.000),2h PBG(OR= 1.098,95%CI 1.065-1.132,P=0.000),HbA1c(OR=1.043,95%CI 1.002-1.084,P=0.038),瘀血阻滞证(OR=1.251,95%CI 1.025-1.528,P=0.028),痰瘀互阻证(OR=1.319,95%CI 1.016-1.712,P=0.038),阴虚燥热证(O R=0.535,95%CI 0.383-0.749,P=0.000)和脾肾阳虚证(OR=1.660,95%CI 1.074-2.565,P=0.022)与糖尿病微血管病变有相关性。病程(OR=1.069,95%CI 1.054-1.086,P=0.000),家族史(OR=1.324,95%CI 1.085-1.615,P=0.006),2hPBG(OR=1.099,95%CI 1.069,P=1.129),阴虚燥热证(OR=0.515,95%C 1 0.345-0.768,P=0.001)和脾肾阳虚证(OR=1.536,95%CI 1.072-2.202,P=0.019)与 DR 有相关性。年龄(OR=0.979,95%CI 0.968-0.989,P=0.000),病程(OR=1.047,95%CI 1.030-1.064,P=0.000),高血压病(OR=1.697,95%CI 1.311-2.196,P=0.000),心血管病(OR=1.345,95%CI 1.053-1.718,P=0.018),脑血管病(OR=1.354,95%CI 1.031-1.777,P=0.029),FBG(OR=1.029,95%C 1 1.001-1.059,P,0.044),2hPBG(OR=1.051,95%CI 1.021-1.082,P=0.001),SBP(OR=1.008,95%CI 1.001-1.014,P=0.015),痰瘀互阻证(OR=1.438,95%CI 1.099-1.881,P=0.015),阴虚燥热证(OR=0.603,95%CI 0.376-0.968,P=0.036),脾肾阳虚证(OR=2.286,95%CI 1.583-3.301,P=0.000)与 DN 有相关性。病程(OR=1.043,95%CI 1.029-1.058,P=0.000),家族史(OR=1.399,95%CI 1.158-1.691,P=0.001),高脂血症(OR=1.328,95%CI 1.049-1.682,P=1.328),心血管病(OR=1.555,95%CI 1.253-1.929,P=0.000),BMI(OR=1.033,95%CI 1.008-1.058,P=0.009),2hPBG(OR=1.093,95%CI 1.064-1.123,P=0.000),痰瘀互阻证(OR=1.386,95%CI 1.088-1.765,P=0.008),阴虚燥热证(OR=0.683,95%CI 0.487-0.957,P=0.027)及脾肾阳虚证(OR=1.541,95%CI 1.060-2.241,P=0.023)与DPN有相关性。由患者临床证型可知位于前三位的是气阴两虚证(58.3%)瘀血阻滞证(43.4%)和痰瘀互阻证(18.5%)。由病程可知:病程≤5年,有52.1%患者出现微血管病变;病程6-10年,有61.8%患者出现微血管病变;病程11-15年,有72.9%患者出现微血管病变,病程16-20年,有77.6%患者出现微血管病变;病程20年,有80.9%患者出现微血管病变。由HbA1c可知,HbA1c水平越高,微血管病变发生率就越高;同样,由B MI可知,BMI水平越高,微血管病变发生率就越高。根据前期研究得到的5个DR危险因素,在2041例T2DM患者中进行匹配,最终纳入符合条件的12对共24例T2DM患者,所有患者之间均无血缘关系。提取的DNA样品全部符合实验要求,将芯片扫描结果比对Hapmap数据,筛选出7691个有效SNP位点,最后筛选出 rs9345283、rs738322、rs2844795、rs1727638、rs16984239、rs12505641、rs12142968、rs11243897、rs11175883这9个12对患者均存在差异的的基因位点,通过NCBI数据库、UCSC数据库、lncRNASNP数据库和HGNC数据库查询到rs738322位点位于PLA2G6基因,rs2844795位点位于TRIM31基因,rs 1727638 位点位于 LINC01626 基因,rs12505641 位点位于 LOC105374524 基因 r s11243897 位点位于 AK8 基因;而 rs9345283 位点、rs16984239 位点、rs12142968位点及rs1 1175883位点未找到所在基因。通过KEGG数据库查找相关基因功能:PLA2G6基因参与炎症调节及脂质代谢,TRIM31基因参与免疫应答调控,AK8基因参与上皮细胞迁移中的负调控,未查找到LOC105374524基因和LINC 01626基因相应的功能。结论:1病程,家族史,高血压病,心血管病,2hPBG,HbA1c,瘀血阻滞证,痰瘀互阻证,阴虚燥热证和脾肾阳虚证为糖尿病微血管病变的危险因素;病程,家族史,2hPBG,阴虚燥热证和脾肾阳虚证是DR的危险因素;年龄,病程,高血压病,心血管病,脑血管病,FBG,2hPBG,SBP,痰瘀互阻证,阴虚燥热证,脾肾阳虚证是DN的危险因素;病程,家族史,高脂血症,心血管病,BMI,2h PBG,痰瘀互阻证,阴虚燥热证及脾肾阳虚证是DPN的危险因素。2考虑PLA2基因多态性可能通过炎症介导、调节胰岛分泌及胰岛细胞功能、调节血管平滑肌收缩功能,参与糖尿病视网膜病变的发生。Trim31基因可能通过活化NLRP3炎性体作用,参与糖尿病视网膜病变的发生。
[Abstract]:Objective: to find out the risk factors associated with diabetic microangiopathy by analyzing the general data, clinical symptoms, TCM syndromes and laboratory tests of diabetic patients, and on the basis of the risk factors of diabetic retinopathy, screening patients' independent susceptibility genes for the future type 2 diabetic microvascular lesions. The in-depth study of prevention and treatment and diabetic retinopathy was provided. Methods: (1) a self compiled questionnaire was used to collect information about the T2DM patients in the endocrinology outpatient department and the inpatient department of Beijing, Shijiazhuang and Baoding during the period from March 2016 to January 2017. The cross section analysis was carried out. The factors were analyzed by Logistic regression analysis. Single factor and multi factor analysis. (2) on the basis of identifying the risk factors of DR in the early stage, the relevant information (including general data, clinical symptoms and laboratory tests) with all the DR patients with all the risk factors of DR (including general data, clinical symptoms and laboratory tests) was matched, and a pair of DR and non DR patients with all relevant information were selected and included in the study. The DNA of the venous whole blood of the patient, using the ILLUMINA human SNP typing detection chip technology, screening 242901 SNP loci of the DR patients to find all the different sites in the group, through the NCBI database (http://www.ncbi.nlm.nih.gov/), the UCSC database (http:// genome.ucsc.edu/), lncR NASNP. According to the library and the HGNC database (http://www.gen enames.org/), the gene was searched for the loci of the heterotopic and the KEGG database (http://www.kegg.jp/) was used to find the gene metabolic pathway. Results: 2041 cases of diabetic patients entered the investigation, including 1318 patients with microangiopathy (734 cases of diabetic retinopathy, only 734 cases of diabetic retinopathy, " There were 138 patients with diabetic retinopathy, 468 patients with diabetic nephropathy, 96 patients with diabetic nephropathy, 1044 patients with diabetic peripheral neuropathy, 379 patients with diabetic peripheral neuropathy and 723 patients without microvascular disease. Single factor Logistic regression analysis showed year Age (OR=1.016,95%CI 1.008-1.023, P=0.000), course of disease (OR=1.071,95%CI 1.005-1.086, P=0.000), family history (OR=1.306,95%CI 1.085-1.573, P=0.005), hypertension (OR=1.651,95%CI 1.376-1.983, P=0.000), hyperlipidemia, cardiovascular disease, cerebrovascular disease 9,95%CI 1.009-1.595, P=0.042), BMI (OR=1.035,95%CI 1.011-1.059, P=0.003), FBG (OR= 1.025,95%CI 1.001-1.050, P=0.045). 95%CI 1.038-1.502, P=0.018), phlegm and stasis syndrome (OR=1.460,95%CI 1.144-1.865, P=0.002), yin deficiency and heat syndrome (OR=0.444,95%CI 0.326-0.606, P=0.000), spleen kidney yang deficiency syndrome (O R=2.335,95%CI 1.549-3.520) and diabetic microvascular disease. O R=1.241,95%CI 1.016-1.517 (P=0.035), hypertension (OR=1.315,95%CI 1.052-1.643, P=0.035), cardiovascular disease (OR=1.522,95%CI 1.204-1.924, P=0.000), 2h PBG, stagnation of blood stasis syndrome, syndrome of stagnation of phlegm and blood stasis syndrome (phlegm and stasis syndrome). OR=1.319,95%CI 1.016-1.712, P=0.038), yin deficiency and heat syndrome (O R=0.535,95%CI 0.383-0.749, P=0.000) and spleen and kidney yang deficiency syndrome (OR=1.660,95%CI 1.074-2.565, P=0.022) are related to diabetic microvascular disease. =1.129), yin deficiency and heat syndrome (OR=0.515,95%C 1 0.345-0.768, P=0.001) and spleen and kidney yang deficiency syndrome (OR=1.536,95%CI 1.072-2.202, P=0.019) are related to DR. Age (OR=0.979,95%CI 0.968-0.989, P=0.000), disease course (OR=1.047,95%CI), high blood pressure disease, cardiovascular disease (1.) 053-1.718, P=0.018), cerebrovascular disease (OR=1.354,95%CI 1.031-1.777, P=0.029), FBG (OR=1.029,95%C 1 1.001-1.059, P, 0.044), 2hPBG (OR=1.051,95%CI 1.021-1.082), syndrome of stagnation of phlegm and stasis, spleen deficiency and heat syndrome OR=2.286,95%CI 1.583-3.301 (P=0.000) has a correlation with DN. The course of disease (OR=1.043,95%CI 1.029-1.058, P=0.000), family history (OR=1.399,95%CI 1.158-1.691, P=0.001), hyperlipidemia (OR=1.328,95%CI 1.049-1.682), cardiovascular disease. G (OR=1.093,95%CI 1.064-1.123, P=0.000), phlegm and blood stasis syndrome (OR=1.386,95%CI 1.088-1.765, P=0.008), yin deficiency and heat syndrome (OR=0.683,95%CI 0.487-0.957, P=0.027) and spleen and kidney yang deficiency syndrome (OR=1.541,95%CI 1.060-2.241) are related. It is known from the patient's bed syndrome that the first three places are Qi Yin two deficiency syndrome (58.3%) stagnation of blood stasis syndrome (43.4%) and the mutual obstruction syndrome of phlegm and stasis (18.5%). It is known from the course of the disease that the course of the disease is less than 5 years, there are 52.1% patients with microvascular lesions, 61.8% patients have microvascular lesions for 6-10 years, 11-15 years in the course of disease, 72.9% patients with microvascular lesions, 77.6% patients with microvascular lesions, 20 years in the course of disease, and microvascular lesions in 80.9% patients. HbA1c shows that the higher the level of HbA1c, the higher the incidence of microvascular lesions; also, the higher the level of BMI, the higher the BMI level, the higher the incidence of microvascular lesions. According to the 5 DR risk factors obtained from the previous study, 2041 cases of T2DM patients were matched, and 12 pairs of T2DM patients were included, and all the patients had no blood. The extracted DNA samples all meet the requirements of the experiment. 7691 effective SNP loci are screened by the results of the chip scanning compared with the Hapmap data. Finally, the rs9345283, rs738322, rs2844795, rs1727638, rs16984239, rs12505641, rs12142968, rs11243897, and rs11175883 are the 9 12 gene loci, which are different in the number of patients. According to the library, the UCSC database, the lncRNASNP database and the HGNC database inquire that the rs738322 loci are located in the PLA2G6 gene, the rs2844795 site is located in the TRIM31 gene, the RS 1727638 loci is located in the LINC01626 gene, and the rs12505641 loci is located in the LOC105374524 gene. 68 loci and RS1 1175883 loci did not find the gene. Through the KEGG database to find the related gene function: PLA2G6 gene involved in the regulation of inflammation and lipid metabolism, TRIM31 gene involved in the regulation of immune response, AK8 gene involved in the negative regulation of epithelial cell migration, and the function of LOC105374524 gene and LINC 01626 gene was not found. Conclusion: 1 The course, family history, hypertension, cardiovascular disease, 2hPBG, HbA1c, stagnation of blood stasis syndrome, phlegm and stasis syndrome, yin deficiency and heat syndrome and spleen kidney yang deficiency syndrome are the risk factors of diabetic microvascular disease; the course of disease, family history, 2hPBG, yin deficiency and heat syndrome and spleen kidney yang deficiency syndrome are risk factors for DR; age, course of disease, hypertension, cardiovascular disease, cerebrovascular disease, FBG, 2h PBG, SBP, mutual obstruction of phlegm and stasis syndrome, yin deficiency and heat syndrome, spleen kidney yang deficiency syndrome are the risk factors of DN; the course of disease, family history, hyperlipidemia, cardiovascular disease, BMI, 2h PBG, phlegm stasis syndrome, yin deficiency and heat syndrome and spleen kidney yang deficiency syndrome are the risk factors of DPN, which can regulate the secretion of islet and the function of islet cell, regulate the function of islet cells and regulate the function of islet cells, and regulate the function of islet cells. The contractile function of vascular smooth muscle and the involvement of.Trim31 gene in diabetic retinopathy may be involved in the pathogenesis of diabetic retinopathy by activating the role of NLRP3 inflammatory body.

【学位授予单位】：中国中医科学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R259;R276.7

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