Genetics Analysis And Traditional Chinese Medicine Clinical Exploration On Mutations In The Thiazide-sensitive Na-Cl Co-transporter Gene (SLC12A3) In Five Families

ObjectiveNa-Cl co-transporter（NCCT） is an important protein to maintain the stability of blood pressure and the the balance of water and salt electrolyte,this study attempts to explore the genetic characteristics of five families which diagnosed with Gitelman’s syndrome based on NCCT gene （SLC12A3） defects. And this study is not only to analysis the traditional Chinese medicine syndrome phenotype about the genetic defect disease above-mentioned, but also to discuss the advantages and countermeasures of traditional Chinese medicine prescription for the treatment of this disease.Method1. Under the informed consent, chose24people from five families to study, extracted peripheral blood genomic DNA; Use the tool Primer Primier5software to design primers, and use PCR product amplification26exons of SLC12A3gene, and PCR product purification, directly sequenced by Sanger method looking forward to find the mutations.2. Collect the linical symptoms and biochemical indicators, then diagnos syndrome by traditional Chinese medicine analysis. Treat five propositus according to Western medicine and Chinese medicine progressively and superimposedly, the Western medicine scheme is: oral postassium chloride for2weeks, and panangin combine with spironolactone or ammonia benzene prescription2weeks later, again after2weeks add Chinese traditional medicine （which is sijunzi decoction and ultimately for five tonga subtraction）, during the time test the potassium levels every2weeks.Results1. Family I propositus A has three mutations:Thr60Met、965-1₉76dell3insl2（small indels mutation）, Ala122Ala（homozygous and silent mutations）; Family II propositus B has three mutations:Asn359Lys、Thr382Met、Arg913Gln; Family â…¢ propositus C has two mutations, respectively is:exon1C179T, causes Thr60Met into methionine mutant （threonine mutation into methionine）; exon24C2782T, causes the CGT to TGT change, leading to Arg928Cys, arginine is replaced by the cysteine, missense mutation; 2. Family IV propositus D by mutation gene sequencing respectively is:exon24C2782T, causes the CGT-TGT change, leading to Arg928Cys, Ala122Ala, which are homozygous silent mutation; exon11C1395T, causes the ACC to ACT, leading to Thr465Thr are homozygous silent mutation; Family V propositus E also have a mutation point:exon17C2129A, causes the TCG-> TAG, causes Ser710Term, where comes out the termination codon, for nonsense mutations;3. During our treatment, the potassium level and symptoms were improved in different stages of these five probands.ConclusionsIn this study there are three GS families for compound heterozygous mutations, mostly missense mutation, two families for homozygous mutations, the way of homozygous mutations was found for the first time. Recently researches at home and abroad have shown that Gitelman’s syndrome in patients with specific link between genotype and phenotype are difficult to determine. The current domestic have not set up the improvement of the clinical diagnosis of gitelman syndrome screening system, lack of large sample data about sodium chlorine transporter defect families, and is unable to make more accurate predictions and assessments about the incidence of Chinese GS temporarily. GS’s confirmed still depends on genetic diagnosis.GS herbal treatment is still lack of conventional prescription, our traditional Chinese medicine treatment based on the sijunzi decoction and buyanghuanwudecoction （BHD）, and we put the prescription as our conventional prescription. We use the Western medicine-Chinese medicine superposition of treatment step by step on five proposituses. Through the observation and record of potassium and other biochemical examination levels and the improvement of clinical symptoms on objects of our study, we find that the treatment effect is good, especially in improving the clinical symptoms effect is remarkable.