Study Of Association Between The First Stage Of CKD With Damp-heat Constitution And ICAM-1 Gene Polymorphism And Expression Of Serum SICAM-1

ObjectiveTo study the association among patients with first stage of CKD with damp-heat constitution and ICAM-1 G241R/K469E polymorphism and sICAM-1 levels in serum. To analyze the differences of ICAM-1 gene polymorphisms and sICAM-1 levels in serum among damp-heat constitution, first stage of CKD and first stage of CKD with damp-heat constitution. So that to compare the influence of sICAM-1 levels in serum among different genotypes.Finally, to discusse clinical significance of diagnosis of early CKD and damp-heat constitution by ICAM-1 gene polymorphism and serum level of sICAM-1 test.Methods1.Patients in inpatient department of nephrology and outpatient of chronic disease, who were in first stage of CKD, would be identified the constitution of TCM and brought into the study if accord with inclusion and exclusion criteria, and divide into first stage of CKD with damp-heat constitution group and first stage of CKD with normal constitution group. Healthy people in medical examination center and outpatient of health management,would be identified the constitution of TCM and brought into the study if accord with inclusion and exclusion criteria, and divide into healthy damp-heat constitution group and healthy normal constitution group.2. All the subjects were collected venous blood samples after informed consent.Each blood sample extracted genomic DNA, then used ultraviolet specrophotometer to test purity and concentration of genomic DNA, these products were amplified by PCR, and verified whether the PCR products were objective fragmented gene by agarose gel electrophoresis,finally sent the products to biological company to sequence,different genotypes of ICAM-1 G241R/K469E could be judged by sequenced atlas.To analyze the difference of frequency of ICAM-1 G241R/K469E genotype and allele in four study groups, as well as study the relationship between the difference and first stage of CKD with damp-heat constitution. In the end, test the population representativeness by Hardy-Weinberg law.3. ELISA method is used to detect serum sICAM-1 level. Then analyze the difference of serum sICAM-1 levels among four groups, study the relationship between the difference and first stage of CKD with damp-heat constitution, as well as the relationship the difference and ICAM-1 G241R/K469E polymorphism.Results1. There were 117 study objects were brought into the study, included 66 cases of first stage of CKD group and 51 cases of healthy group. Three kinds of disease category in first stage of CKD group including primary glomerulonephritis(52 cases), LN(8 cases), chronic pyelonephritis (6 cases). The average age was similar in four groups, age factor didn’t affect the study.2. There was no statistical difference of ICAM-1 G241R/K469E genotypes of all subjects between actual observed frequency number and theoretical frequency number, it conformed to the Hardy-Wei nberg law(P>0.05).3. In all study objects, genotype frequencies of ICAM-1 G241Rwere GG0.692, GR 0.308, RR didn’t been detected, the G/R allele frequencies was 0.846/0.154. Different genotype and allele frequencies were same in four study groups.4. In all study objects, genotype frequencies of ICAM-1 K469Ewere KKO.59, KE 0.376,EE 0.034,the K/E allele frequencies was 0.778/0.222. In patients with first stage of CKD, KE/EE genotype frequencies of whom had damp-heat constitution were higher than whom had normal constitution(0.455 vs 0.273, 0.121 vs 0), but KK genotype frequencies was lower (0.424 vs 0.727),E allele frequencies of whom had damp-heat constitution were higher than whom had normal constitution(0.348 vs 0.136), but K allele frequencies was lower(0.652 vs 0.864). In people had damp-heat constitution, KE/EE genotype frequencies of whom with first stage of CKD were higher than healthy people(0.455 vs 0.32, 0.121 vs 0), but KK genotype frequencies was lower(0.424 vs 0.68),E allele frequencies of whom with first stage of CKD were higher than healthy people (0.348 vs 0.16), but K allele frequencies was lower (0.652 vs 0.84).5. Population distribution of serum sICAM-1 levels was not all the same in four study groups. First stage of CKD with damp-heat constitution group had higher expression of serum sICAM-1 than first stage of CKD with normal constitution group. First stage of CKD with damp-heat constitution group had higher expression of serum sICAM-1 than healthy normal constitution group. Healthy damp-heat constitution group had higher expression of serum sICAM-1 than first stage of CKD with normal constitution group. Healthy damp-heat constitution group had higher expression of serum sICAM-1 than healthy normal constitution group.6. In four study groups, serum sICAM-1 levels were same in GG, GR genotypes of ICAM-1 G241R. But in first stage of CKD with damp-heat constitution group, KE genotype of ICAM-1 K469E had higher expression of serum sICAM-1 than KK genotype. In other groups, serum sICAM-1 levels were same in KK, KE genotypes of ICAM-1 K469E.Conclusion1. There was no statistical difference of ICAM-1 G241R/K469E genotypes of all subjects between actual observed frequency number and theoretical frequency number, it conformed to the Hardy-Weinberg law, the sample size in this study can represent one population.2. In all study objects, most of ICAM-1 G241R, K469E genotypes are GG or GR and KK or KE, most of alleles are G and K. It has same distribution of ICAM-1 G241R polymorphism in four study groups, and didn’t show the correlation of damp-heat constitution and early CKD. E allele of ICAM-1 K469E may be one of risk factor to damp-heat constitution and early CKD, people with KE or KK genotype may show the susceptibility to damp-heat syndrome and CKD.3. Damp-heat constitution maybe one of independent risk factors to cause higher expression of serum sICAM-1, but first stage of CKD didn’t show the correlation of serum sICAM-1 higher expression.4. ICAM-1 G241R genotypes and serum sICAM-1 levels had no obvious correlation. E allele of ICAM-1 K469E may have enhancement to serum sICAM-1 expression, it may be one of genetic risk factors to cause higher expression of serum sICAM-1, so KE genotype heterozygote that contains E allele also have enhancement to serum sICAM-1 expression.