Investigating The Correlation Between ENOS Gene Single Nucleotide Polymorphisms And The Chinese Medical Evidence Of Systemic Lupus Erythematosus In Hainan

Objective:By analyzing the mononuclear polymorphic changes of e NOS gene in healthy individuals,SLE patients with yin deficiency and internal heat syndrome,and spleen kidney yang deficiency syndrome in Hainan region,potential gene loci that can be used to assist in syndrome differentiation are identified.Exploring the objective basis of TCM syndrome differentiation in SLE from a micro level,enriching the theoretical connotation of objectification of TCM syndrome differentiation and treatment,and accumulating data for the objectification research of SLE syndrome.Method:133 healthy controls and 133 SLE patients in Hainan(58 patients with yin deficiency and internal heat syndrome and 75 patients with spleen and kidney yang deficiency syndrome)were selected as the research subjects,and their clinical basic information,traditional Chinese medicine four diagnostic information,laboratory tests,and blood samples were collected.The SNap Shot genotyping technique was used to determine the single nucleotide polymorphisms at the rs3918181,rs1808593,and rs2070744 loci of the e NOS gene in the study subjects.Use SPSS25.0 software to statistically analyze the correlation between these three loci of e NOS gene in healthy controls,SLE patients with yin deficiency internal heat syndrome,and spleen kidney yang deficiency patients.Results:1.This study collected 58 SLE patients with yin deficiency and internal heat syndrome,and 75 SLE patients with spleen and kidney yang deficiency.The top 10 syndromes of yin deficiency and internal heat syndrome were: lack or absence of tongue coating,fever,yellow tongue coating,light red tongue,pulse number,fine pulse,night sweat,hot flashes,red tongue,and joint fixed pain.The top 10 syndromes of spleen and kidney yang deficiency syndrome include: tongue tooth marks,edema,pale tongue,weak pulse,fatigue,poor appetite,yellow tongue coating,white tongue coating,skin erythema,and abdominal distension.2.In the comparison of laboratory indicators between SLE patients with yin deficiency internal heat syndrome and spleen kidney yang deficiency syndrome,the increase in blood creatinine accounted for 100.0% in the yin deficiency internal heat group and less than 2.7% in the spleen kidney yang deficiency group.There was a significant difference between the two groups(P<0.001);The increase in blood urea nitrogen accounted for less than 1.7% in patients with yin deficiency and internal heat syndrome,and 10.7% in patients with spleen and kidney yang deficiency syndrome.The difference between the two groups was statistically significant(P<0.05).3.The HWE-P values for the e NOS gene rs3918181,rs1808593 and rs2070744 loci were all greater than 0.05.4.The genotype and allele frequency distribution of the e NOS gene rs1808593 locus showed significant differences between healthy individuals and SLE patients(P<0.01),and there were also statistical differences between the GT and TT genotypes between the two groups(P<0.01).The OR of GT genotype is 0.361,95%CI is 0.211-0.616;The OR of TT genotype is 2.445,95% CI is 1.461～4.091;The OR of allele G is 0.552,95% CI is 0.358-0.852;The OR of allele T is 1.811,and 95% CI is 1.174～2.795.The genotype and allele frequency of rs3918181 and rs2070744 loci of e NOS gene had no statistical difference between healthy people and SLE patients(P>0.05).5.In the comparison between healthy people and SLE patients with yin deficiency and internal heat syndrome,there was no statistical difference in the distribution of genotype and allele frequency of the e NOS gene rs3918181,rs1808593 and rs2070744 loci between the two groups(P>0.05).In the comparison between healthy individuals and SLE patients with spleen and kidney yang deficiency syndrome,the genotype and allele frequency distribution of the e NOS gene at rs1808593 and rs2070744 loci showed significant differences between the two groups(P<0.05).And the GT and TT genotypes at rs1808593 locus and the CT and TT genotypes at rs2070744 locus also showed statistical differences between the two groups(P<0.05).The OR of the GT genotype at rs1808593 locus is 0.199,95% CI is0.094～0.421,the OR of the TT genotype is 3.805,95% CI is 1.940～7.461,the OR of the G allele is 0.413,95% CI is 0.235～0.727,the OR of the T allele is 2.420,95% CI is 1.375～4.261;The OR of the CT genotype at rs2070744 locus is 0.393,95% CI is0.170～0.907,the OR of the TT genotype is 2.722,95% CI is 1.230～6.024,the OR of the C allele is 0.392,95% CI is 0.190～0.808,the OR of the T allele is 2.551,and 95%CI is 1.238～5.255.6.In the comparison of SLE with yin deficiency and internal heat syndrome and spleen kidney yang deficiency syndrome,the genotype and allele frequency distribution of e NOS gene rs3918181 locus had no statistical difference between the two groups(P>0.05),while the genotype and allele frequency distribution of rs1808593 and rs2070744 locus had statistical difference between the two groups(P<0.05).And the GT and TT genotypes of rs1808593 locus and CT,TT genotypes of rs2070744 locus The TT genotype also showed statistical differences between the two groups(P<0.05).OR of GT genotype at rs1808593 locus=0.271,95%CI=0.115～0.637,OR for TT genotype=2.863,95% CI=1.308～6.269,OR for G allele genotype=0.496,95% CI=0.256～0.962,OR for T allele genotype=2.015,95%CI=1.039～3.906;The OR of the CT genotype at the rs2070744 locus is 0.375,95%CI is 0.145～0.969,the OR of the TT genotype is 3.041,95% CI is 1.240～7.457,the OR of the C allele is 0.343,95% CI is 0.154～0.765,the OR of the T allele is 2.917,and 95% CI is 1.308～6.506.Conclusion:1.Elevated blood creatinine and elevated blood urea nitrogen may be laboratory indicators that differentiate between SLE yin deficiency internal heat syndrome and spleen kidney yang deficiency syndrome.2.In healthy individuals and SLE patients in Hainan region,the rs3918181,rs1808593,and rs2070744 loci of the e NOS gene all met the Hardy-Weinberg genetic balance test.3.In the comparison between healthy individuals and SLE,the rs1808593 locus of the e NOS gene is associated with susceptibility to SLE,and genotype GT and allele G are protective factors for SLE susceptibility,while genotype TT and allele T are risk factors for SLE susceptibility.4.In the comparison between healthy individuals and SLE with spleen and kidney yang deficiency syndrome,rs1808593 and rs2070744 of the e NOS gene are associated with the susceptibility of SLE with spleen and kidney yang deficiency syndrome.The GT and CT genotypes,G and C alleles at these two loci are protective factors for SLE with spleen and kidney yang deficiency syndrome,while TT genotype and T allele are risk factors for SLE with spleen and kidney yang deficiency syndrome.5.In the comparison between SLE Yin Deficiency Internal Heat Syndrome and Spleen Kidney Yang Deficiency Syndrome,the rs1808593 and rs2070744 loci of the e NOS gene may be reference indicators for distinguishing SLE Yin Deficiency Internal Heat Syndrome and Spleen Kidney Yang Deficiency Syndrome.The GT and CT genotypes,G and C alleles at these two loci are protective factors for SLE Spleen Kidney Yang Deficiency Syndrome,while the TT and T alleles are risk factors for SLE Spleen Kidney Yang Deficiency Syndrome.