IgA腎病濕熱質(zhì)人群與IL-10基因啟動子及ICAM-1基因位點多態(tài)性關系研究
發(fā)布時間：2019-03-29 17:09
【摘要】：目的：探討IgA腎病濕熱體質(zhì)與白介素10(IL-10)-1082/-819/-592位點及細胞粘附因子1 (ICAM-1) G241R/K469E位點多態(tài)性之間的關系。找出IgA腎病及濕熱體質(zhì)的遺傳易感基因及兩者之間的關系,從而為更好地運用中西醫(yī)結(jié)合的理論防治IgA腎病提供實驗依據(jù)。方法：在住院部及體檢中心收集志愿者,在符合納入標準并除外排除標準的前提下對研究對象進行中醫(yī)體質(zhì)評估并將其分成4組,分別是：IgA腎病濕熱質(zhì)組,IgA腎病平和質(zhì)組,健康濕熱質(zhì)組及健康平和質(zhì)組。抽取其靜脈血3m1以提取DNA。將提取到的DNA進行PCR擴增,得到基因片段后電泳以檢測該片段是否為目的基因,送上海生工公司測序。最后通過測序鋒圖讀取上述五個位點的基因型及等位基因頻率,并運用SPSS19.0軟件進行統(tǒng)計學分析,找出各組之間的差異基因。結(jié)果：1.在納入的98例研究對象中,IgA腎病濕熱質(zhì)組29例,IgA腎病平和質(zhì)組21例,健康濕熱質(zhì)組22例,健康平和質(zhì)組26例,各組之間年齡、性別無統(tǒng)計學差異,說明各組年齡及性別基線齊。2.對IL-10-1082/-891/-592位點及ICAM-1-G241R/K469E位點的基因頻率進行Hardy-Weinberg遺傳平衡檢驗時,計算出IL-10中三個位點基因型的p值0.05,說明具有較好的群體代表性。而ICAM-1中的兩個位點計算出的p值分別是0.039和0.043,均0.05,說明其群體代表性不太理想。3.對上述5個位點基因型及等位基因進行分析得出IL-10-1082位點在四組之間存在統(tǒng)計學差異,而其余位點各組之間無差異。而IL-10-1082位點基因型主要以AA為主(87.8%),GG等位基因未檢測出,等位基因主要以A為主(93.9%)。4.進一步對各組之間IL-10-1082位點基因進行分析,發(fā)現(xiàn)全部IgA腎病人群(不區(qū)分體質(zhì)類型)AA基因型及A等位基因比健康人群要高(94%：81.3%和97%：90.6%)；而在濕熱人群中,IgA腎病人群比健康人群的從基因頻率及A等位基因同樣要高(100%：85.7%和100%：92.9%)；在健康人群當中,濕熱體質(zhì)人群比平和體質(zhì)人群其GG基因型及G等位基因要高(36.4%：3.8%和18.2%：1.9%)。結(jié)論：1.IL-10基因啟動子-1082位點中的AA基因型及A等位基因可能是IgA腎病的遺傳易感因素,具有該基因型的人群更容易得IgA腎病。2.在濕熱體質(zhì)人群中,IgA腎病之人其AA基因頻率及A等位基因比健康人群會更高,而在平和體質(zhì)人群中,該位點在IgA腎病與健康人群之間并未存在差異,這說明IgA腎病可能與濕熱體質(zhì)存在一定相關性。3.在健康人群中,濕熱體質(zhì)人群GG基因頻率反而會高些,而在IgA腎病中卻無此現(xiàn)象。這說明G等位基因可能是健康濕熱質(zhì)的危險因素,而IgA腎病始終與AA基因型及A等位有關。
[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.
【學位授予單位】：廣州中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R692.31
本文編號：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.
【學位授予單位】：廣州中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R692.31
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