Tgf-¦Â1 And Ras Gene Polymorphism And Lumbar Disc Herniation Tcm Syndromes Related Research

Objective:To investigate the relationship between transforming growth factorβ1(TGF-β1), renin-angiotensin system(RAS)related gene:angiotensin-converting enzyme(ACE), angiotensinⅡtype 1 receptor(AT1R)gene polymorphisms and the traditional Chinese medicine syndrome of lumbar intervertebral disk protrusion(LIDP),search for the susceptible gene of the traditional Chinese medicine syndrome of LIDP,explore the possibility of gene faclitilized traditional Chinese medicine syndrome and provide basis on further research of traditional Chinese medicine genetic diagnosis and therapy of LIDP.Methods:1.According to the diagnosis standards of "Lumbar intervertebral disc protrusion" made by Hu you-gu and "Practical Orthopaedics"made by Lu yu-pu et al,Combining patient selection inclusion and exclusion criteria,selecting 118 LIDP patients at random,as cases group, including 63 Han and 55 Zhuang people.At the same time,118 cases are typed into 38 patients of blood stasis pattern,29 patients of cold hygrosis pattern,31 patients of Deficiency of both Liver and Kidney pattern and 20 patients of Dampness and Heat pattern by industry standard approved in 1994 and "new clinical research guiding principle of tradition Chinese medicine cure lumbar intervertebral disc protrusion" drafted by Chinese people public nation health forum in 2002.119 cases were from health examination center as healthy control group,including 64 for Han,55 for Zhuang,and matched with cases group by age gender etal2.Age gender,nation character,waist on-load enery(occupation)and the family history of objects were collected in interview.Body mass index,smoking index were recorded and the relevance subjects such as the level of intervertebral disc degeneration(IDD)were evaluated by MRI test.3.2ml vessel blood of each sample was taken to test the single nucleotide polymorphism(SNP)of TGF-β1 -509,TGF-β1 -800,ACE and AT1R gene A1166C.The technology of Polymerase Chain Reaction-Restriction Fragment Length Polymorphisms(PCR-RFLP)was applied.Main experiment courses included the extraction of peripheral blood,PCR amplification,restriction enzyme digestion,and so on.4 Genotypic frequency and allele frequency were calculated by frequency counter,comparison of genotype and allele frequenc between two groups were taken by x~2 test,comparison of mean between two groups by t test,odds ratio,interaction were analyzed by logistic regression,etal.All statistics were analyzed by SPSS 13.0 soft package.Results:1.DNA PCR amplification of TGF -β1 -509,TGF -β1 -800,ACE and AT1R gene A1166C were welldone.PCR amplification products of TGF-509 gene were digested into three genotypes such as CC,CT and TT type by Saul restrict enzyme,TGF-800 allele amplification products were digested by Tsp45I,only one genotype was detected,homozygous GG.ACE was genotyped intoⅡ,DD and DI types;AT1R gene was genotyped into AA,AC and CC by DdeI.2.The gene polymorphism of TGF-β1 -800 G/A in patient group have no different significance with that in control group,there was significant differents of polymorphism of TGF -β1 -509 C/T and AT1R1166A/C between patient group and control group,but there is no obvious difference between the group.It was showed that there was significant difference between ACE I/D polymorphism in patient group and the control group(P＜0.05).TGF-β1-509 C/T,ACE I/D and AT1R1166 A/C polymorphisms was relevant to IDD.There was signicant between two groups of AT1R1166 genotype distribution(P＜0.05).3.It is indicated that polymorphism of TGF-β1-509,ACE,AT1RA1166C and the serious waist onload enery poses could increase the risk of LIDP.4.The frequency of TGF-β1 CT genotype in blood stasis pattern of LIDP is higher than the non-blood stasis pattern;The TGF-β1 genotype and the frequency of C/T allelic gene in cold hygrosis pattern of LIDP compared with Deficiency of both Liver and Kidney pattern have the significant difference;The TGF-β1 genotype and the frequency of C/T allelic gene in cold hygrosis pattern have the significant difference;The frequency of ACE geneⅡgenotype in Deficiency of both Liver and Kidney pattern are higher than the non-Deficiency of both Liver and Kidney pattern;The frequency of AT1R1166 AC genotype in Zhuang is higher than Han in the cases group,and the patient with cold hygrosis pattern is higher than syndrome of blood stasis pattern. 5.The patient with blood stasis pattern of the LIDP of TGF-β1 -509 CT carrier and ACE ID carrier as well were in higher risk;The patient with cold hygrosis pattern of the LIDP with AT1R1166 gene AC carrier,TGF-β-509 CC carrier were in higher risk than LIDP with cold hygrosis pattern;The patients with Deficiency of both Liver and Kidney pattern of LIDP, and ACE geneⅡcarrier were in higher risk.Conclusions1.The polymorphisms of TGF-β1-509,ACE and AT1RA1166C are not the single risky factor to LIDP,The family history of Low Back Pain,body figure and waist onload enery(occupied)were the main risky factors to LIDP.2.The polymorphisms of TGF-β1- 509,ACE and AT1RA1166C are relevant to IDD.People who carried TGF-β1-509 CT,ACE DD and AT1R1166 AC might have more IDD risk. That is to say,the TGF-509 adiponectin gene and the RAS relevant gene might have relation to the progress of IDD,ACE gene was correlated with the occurrence of LIDP.3.TGF-β1 gene-509 adiponectin CT genotype is susceptible gene of syndrome of blood stasis pattern of LIDP.Serious waist unload enery is one of the risk factors of syndrome of blood stasis pattern of LIDP.4.ACEⅡpolymorphism is susceptible gene of Deficiency of both Liver and Kidney pattern of LIDP.5.AT1R1166 AC genotype and TGF-β1 -509 CC genotype carrier could increase the risk of the cold hygrosis pattern of LIDP.6.TGF-β1-509 CT genotype was dominant in Blood stasis pattern of LIDP,and these IDD carrying with TGF-β1-509 CT genotype are much more serious,whether there was revelance between LIDP and IDD was needed further research.7.The ACE I/D polymorphism and AT1R1166 A/C polymorphism in Han of Guangxi and Zhuang are susceptible gene sign of LIDP.8.There is no revelance to be found bewteen TGF-β1-800 C genotype and LIDP.The subject provides some experiment and clinic foundation for The TCM molecular syndrome of LIDP and offers basis on the further research.Wether relevant mechanism and some indexes could become standards of the syndrome of LIDP should enlarge the sample size to further reseach.