Research On Vitamin D And Tuberculosis Incidence And Its Adjuvant Therapy

Objective:Determine the serum 25-hydroxyvitamin D concentration and the polymorphism of vitamin D receptor gene and analysis the relevance between 25-hydroxyvitamin D concentration,the genotype of vitamin D receptor with tuberculosis.Methods: We enrolled 155 tuberculosis patients and 109 healthy volunteers and measured the serum25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 of them by isotopedilution ultra-performance liquid chromatographytandem mass spectrometry methods.We got 25-hydroxyvitamin D level and defined 25-hydroxyvitamin D level less than 12ng/ml as vitamin D deficiency,25-hydroxyvitamin D level more than 12ng/ml and less than 20ng/ml as low vitamin D status,and 25-hydroxyvitamin D level more than 20ng/ml as vitamin D sufficient.The patients were divided into vitamin D deficiency group with tuberculosis,low vitamin D status group with tuberculosis,vitamin sufficient group with tuberculosis and the control group was divided into vitamin D deficiency control group,low vitamin D status control group,vitamin D sufficient control group for statistical analysing.We collected the general information,the clinical information and laboratory results of the tuberculosis patients and healthy volunteers.We extracted genomic DNA from the blood of patients and healthy volunteers and determined VDR(vitamin D receptor)gene genotypes by polymerasechain reaction-restriction fragment length polymorphism(PCR-RFLP)analysis methods.Combined the 25-hydroxyvitamin D level and VDR gene genotypes,we analysised the relevance between them with tuberculosis and explored the possible immune mechanism of vitamin D acting in tuberculosis.Results:(1)We enrolled 155 tuberculosis patients as case group and 109 healthyvolunteers as control group and after compairing with the two groups,we found that they were equal in sex and age.(2)The mean serum25-hydroxyvitamin D concentration was 9.80�4.87 ng/ml in the case group and 20.72�7.28 ng/ml in the control group,when compaired with the two group the mean serum 25-hydroxyvitamin D concentration in the case group was 10.92 ng/ml higher than that of control group and the difference was statistically significant.(3)We compaired the polymorphisms and frequencies of VDR gene of case group and control group by Chi-square test and found that there was no relevance between VDR genotypes and gene frequencies and tuberculosis.(4)We compaired the mean serum25-hydroxyvitamin D concentration of diferent genotypes of case group and control group and found that the mean serum 25-hydroxyvitamin D concentration was 8.12 �4.69 ng/ml of FF genotype patients and 10.76�6.04 ng/ml of the Ff genotype patients,the mean serum 25-hydroxyvitamin D concentration of FF genotypewas 2.64 ng/ml higher than that of FF genotype patients,the difference was statistically significant.Conclusion : We found there was a strong relevance between serum25-hydroxyvitamin D concentration with tuberculosis,also there is an intrinsic relevance between serum 25-hydroxyvitamin D concentration.Further research is need to reveal the mechanism of vitamin D and to evaluate the adjuvant effect of vitamin D.Objective To establish the chronic guinea pig model of tuberculosis and to evaluate the role of adjuvant therapy with Vitamin D.Methods Forty-eight guinea pigs infected with mycobacterium tuberculosis standard strain(H37Rv)were randomly divided into blank control group,daily regular dosage of vitamin D group,single high dosage of vitamin D group taken every other week,isoniazid and rifampicin group,isoniazid and rifampicin plus daily regular dosage of vitamin D group,and isoniazid and rifampicin plus high dosage of vitamin D taken every other week group according to different treatment plans.There were 8Guinea Pigs in each group.4 weeks after infection,we initiated treatment.4 guinea pigs in each group were weighed,sacrificed,and dissected 4 weeks and 8 weeks after treatment.Later we counted the Colony Forming Unit(CFU).We also performed the pathological examination in lungs and spleens.Results Our model animals had good tolerance and there was no death in each group throughout the infection and treatment.When compared to the control groups,the CFU counts of lungs and spleens of the guinea pigs did not decline,and the pathological changes of lungs and spleens did not improve in the two groups treated with vitamin D alone.Besides,after 4 weeks of treatment the CFU of lungs and spleens were4.98�0.26 lg CFU and 4.02�0.03 lg CFU in the group of isoniazid and rifampicin and 4.39�0.11 lg CFU and 2.30�0.43 lg CFU in the group of isoniazid and rifampicin plus a single high dosage of vitamin D.When compared the group of isoniazid and rifampicin plus a single high dosage of vitamin D every other week with the group of isoniazid and rifampicin alone,the CFU counts decreased by 0.59 lg CFU in lungs and 1.72 lg CFU in spleens.After 8 weeks of treatment the CFU of lungs and spleens were3.73�0.23 and 2.26�0.24 lg CFU in the group of isoniazid and rifampicin,and the CFU of lungs were 3.21�0.23 lg CFU and spleens were amicrobic in the group of isoniazid and rifampicin plus a single high dosage of vitamin D.When compared the group of isoniazid and rifampicin plus a single high dosage of vitamin D every other week with the group of isoniazid and rifampicin alone,the CFU counts decreased by 0.52 lg CFU in lungs and 2.26 lg CFU in spleens.Also the pathological changes of spleens in the group of isoniazid and rifampicin plus a single high dosage of vitamin D after4 weeks of treatment improved and so as the lungs in the same group but of 8 weeks of treatment.The CFU of spleens were 2.36�0.10 lg CFU in the group of isoniazid and rifampicin plus daily regular dosage of vitamin D after 4 weeks of treatment.When compared the group of isoniazid and rifampicin plus daily regular dosage of vitamin D with isoniazid and rifampicin alone,the CFU of spleens decreased by 1.66 lg CFU after 4 weeks of treatment.Conclusion In the chronic tuberculosis model of guinea pig,vitamin D shows the effect as an adjuvant therapy when combined with isoniazid and rifampicin.It can be furtherstudiedfor the clinical practice.