IgA肾病湿热质人群与IL-10基因启动子及ICAM-1基因位点多态性关系研究
发布时间：2019-03-29 17:09
【摘要】：目的：探讨IgA肾病湿热体质与白介素10(IL-10)-1082/-819/-592位点及细胞粘附因子1 (ICAM-1) G241R/K469E位点多态性之间的关系。找出IgA肾病及湿热体质的遗传易感基因及两者之间的关系,从而为更好地运用中西医结合的理论防治IgA肾病提供实验依据。方法：在住院部及体检中心收集志愿者,在符合纳入标准并除外排除标准的前提下对研究对象进行中医体质评估并将其分成4组,分别是：IgA肾病湿热质组,IgA肾病平和质组,健康湿热质组及健康平和质组。抽取其静脉血3m1以提取DNA。将提取到的DNA进行PCR扩增,得到基因片段后电泳以检测该片段是否为目的基因,送上海生工公司测序。最后通过测序锋图读取上述五个位点的基因型及等位基因频率,并运用SPSS19.0软件进行统计学分析,找出各组之间的差异基因。结果：1.在纳入的98例研究对象中,IgA肾病湿热质组29例,IgA肾病平和质组21例,健康湿热质组22例,健康平和质组26例,各组之间年龄、性别无统计学差异,说明各组年龄及性别基线齐。2.对IL-10-1082/-891/-592位点及ICAM-1-G241R/K469E位点的基因频率进行Hardy-Weinberg遗传平衡检验时,计算出IL-10中三个位点基因型的p值0.05,说明具有较好的群体代表性。而ICAM-1中的两个位点计算出的p值分别是0.039和0.043,均0.05,说明其群体代表性不太理想。3.对上述5个位点基因型及等位基因进行分析得出IL-10-1082位点在四组之间存在统计学差异,而其余位点各组之间无差异。而IL-10-1082位点基因型主要以AA为主(87.8%),GG等位基因未检测出,等位基因主要以A为主(93.9%)。4.进一步对各组之间IL-10-1082位点基因进行分析,发现全部IgA肾病人群(不区分体质类型)AA基因型及A等位基因比健康人群要高(94%：81.3%和97%：90.6%)；而在湿热人群中,IgA肾病人群比健康人群的从基因频率及A等位基因同样要高(100%：85.7%和100%：92.9%)；在健康人群当中,湿热体质人群比平和体质人群其GG基因型及G等位基因要高(36.4%：3.8%和18.2%：1.9%)。结论：1.IL-10基因启动子-1082位点中的AA基因型及A等位基因可能是IgA肾病的遗传易感因素,具有该基因型的人群更容易得IgA肾病。2.在湿热体质人群中,IgA肾病之人其AA基因频率及A等位基因比健康人群会更高,而在平和体质人群中,该位点在IgA肾病与健康人群之间并未存在差异,这说明IgA肾病可能与湿热体质存在一定相关性。3.在健康人群中,湿热体质人群GG基因频率反而会高些,而在IgA肾病中却无此现象。这说明G等位基因可能是健康湿热质的危险因素,而IgA肾病始终与AA基因型及A等位有关。
[Abstract]:Aim: to investigate the relationship between the damp-heat constitution of IgA nephropathy and the polymorphism of interleukin-10 (IL-10)-1082 / 819 / 592 site and cell adhesion factor-1 (ICAM-1) G241R/K469E site. To find out the genetic susceptibility genes of IgA nephropathy and damp-heat constitution and the relationship between them, so as to provide experimental basis for better use of the theory of combination of traditional Chinese and western medicine to prevent and treat IgA nephropathy. Methods: volunteers were collected from the inpatient department and the physical examination center. On the premise of meeting the inclusion criteria and excluding the exclusion criteria, the subjects were divided into four groups: IgA nephropathy damp-heat group, IgA nephropathy level and plasma group, and the subjects were divided into 4 groups: IgA nephropathy damp-heat group, IgA nephrosis level group and plasma group, and the subjects were divided into 4 groups. Healthy damp-heat group and healthy pacification group. Extraction of 3m1 from venous blood to extract DNA. The extracted DNA was amplified by PCR, and the gene fragment was obtained by electrophoresis in order to detect whether the fragment was the target gene and sent to Shanghai Shenggong Company for sequencing. Finally, the genotype and allele frequencies of the above five loci were read by sequencing frontograph, and statistical analysis was carried out by using SPSS19.0 software to find out the different genes among each group. Results: 1. Among the 98 subjects, there were 29 cases in IgA nephropathy group, 21 cases in IgA nephropathy group, 22 cases in healthy damp-heat group, 26 cases in healthy pacification group. There was no significant difference in age and sex among each group. Description of age and gender baseline alignment in each group 2. When the gene frequencies of IL-10-1082/-891/-592 locus and ICAM-1-G241R/K469E locus were tested by Hardy-Weinberg, the p value of the genotypes of the three loci in IL-10 was calculated to be 0.05, which indicated that the gene frequencies of the three loci were well represented in the population. The P values of the two loci in ICAM-1 were 0.039 and 0.043, respectively, both of which were 0.05, indicating that the population representation of the two loci was not ideal. 3. The genotypes and alleles of the above five loci were analyzed and the results showed that there were statistical differences among the four groups of IL-10-1082 loci, but there was no difference among the other loci among the four groups. The main genotype of IL-10-1082 locus was AA (87.8%), GG allele was not detected) and allele A was mainly A (93.9%). Further analysis of IL-10-1082 locus gene showed that AA genotype and A allele were higher in all patients with IgA nephropathy than those in healthy population (94% / 81.3% and 97% / 90.6%, respectively), and the genotype and A allele were higher in all patients with IgA nephropathy than those in healthy population (94% / 81.3% and 97% / 90.6%). In the damp-heat population, the frequency and A allele of IgA nephropathy were also higher than those of healthy people (100% vs 85.7% and 100% / 92.9%). In the healthy population, the GG genotype and G allele were higher in the damp-heat constitution group than those in the mild constitution group (36.4% vs 3.8% and 18.2% / 1.9%). Conclusion: the AA genotype and A allele in the promoter of 1.IL-10 gene at-1082 locus may be the genetic susceptibility factors of IgA nephropathy. The population with this genotype is more likely to develop IgA nephropathy. In the population with damp-heat constitution, the frequency of AA gene and allele A in IgA nephropathy group were higher than those in healthy population, but there was no difference between IgA nephropathy and healthy population in mild constitution group. This indicates that there may be some correlation between IgA nephropathy and damp-heat constitution. 3. In healthy population, the frequency of GG gene was higher in damp-heat constitution group, but not in IgA nephropathy. This suggests that G allele may be a risk factor for healthy damp-heat, and IgA nephropathy is always associated with AA genotype and A allele.
【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R692.31
本文编号：2449723
[Abstract]:Aim: to investigate the relationship between the damp-heat constitution of IgA nephropathy and the polymorphism of interleukin-10 (IL-10)-1082 / 819 / 592 site and cell adhesion factor-1 (ICAM-1) G241R/K469E site. To find out the genetic susceptibility genes of IgA nephropathy and damp-heat constitution and the relationship between them, so as to provide experimental basis for better use of the theory of combination of traditional Chinese and western medicine to prevent and treat IgA nephropathy. Methods: volunteers were collected from the inpatient department and the physical examination center. On the premise of meeting the inclusion criteria and excluding the exclusion criteria, the subjects were divided into four groups: IgA nephropathy damp-heat group, IgA nephropathy level and plasma group, and the subjects were divided into 4 groups: IgA nephropathy damp-heat group, IgA nephrosis level group and plasma group, and the subjects were divided into 4 groups. Healthy damp-heat group and healthy pacification group. Extraction of 3m1 from venous blood to extract DNA. The extracted DNA was amplified by PCR, and the gene fragment was obtained by electrophoresis in order to detect whether the fragment was the target gene and sent to Shanghai Shenggong Company for sequencing. Finally, the genotype and allele frequencies of the above five loci were read by sequencing frontograph, and statistical analysis was carried out by using SPSS19.0 software to find out the different genes among each group. Results: 1. Among the 98 subjects, there were 29 cases in IgA nephropathy group, 21 cases in IgA nephropathy group, 22 cases in healthy damp-heat group, 26 cases in healthy pacification group. There was no significant difference in age and sex among each group. Description of age and gender baseline alignment in each group 2. When the gene frequencies of IL-10-1082/-891/-592 locus and ICAM-1-G241R/K469E locus were tested by Hardy-Weinberg, the p value of the genotypes of the three loci in IL-10 was calculated to be 0.05, which indicated that the gene frequencies of the three loci were well represented in the population. The P values of the two loci in ICAM-1 were 0.039 and 0.043, respectively, both of which were 0.05, indicating that the population representation of the two loci was not ideal. 3. The genotypes and alleles of the above five loci were analyzed and the results showed that there were statistical differences among the four groups of IL-10-1082 loci, but there was no difference among the other loci among the four groups. The main genotype of IL-10-1082 locus was AA (87.8%), GG allele was not detected) and allele A was mainly A (93.9%). Further analysis of IL-10-1082 locus gene showed that AA genotype and A allele were higher in all patients with IgA nephropathy than those in healthy population (94% / 81.3% and 97% / 90.6%, respectively), and the genotype and A allele were higher in all patients with IgA nephropathy than those in healthy population (94% / 81.3% and 97% / 90.6%). In the damp-heat population, the frequency and A allele of IgA nephropathy were also higher than those of healthy people (100% vs 85.7% and 100% / 92.9%). In the healthy population, the GG genotype and G allele were higher in the damp-heat constitution group than those in the mild constitution group (36.4% vs 3.8% and 18.2% / 1.9%). Conclusion: the AA genotype and A allele in the promoter of 1.IL-10 gene at-1082 locus may be the genetic susceptibility factors of IgA nephropathy. The population with this genotype is more likely to develop IgA nephropathy. In the population with damp-heat constitution, the frequency of AA gene and allele A in IgA nephropathy group were higher than those in healthy population, but there was no difference between IgA nephropathy and healthy population in mild constitution group. This indicates that there may be some correlation between IgA nephropathy and damp-heat constitution. 3. In healthy population, the frequency of GG gene was higher in damp-heat constitution group, but not in IgA nephropathy. This suggests that G allele may be a risk factor for healthy damp-heat, and IgA nephropathy is always associated with AA genotype and A allele.
【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R692.31
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