| Objective:To determine the relations between macrophage migration inhibitory factor (MIF) gene promoter-173G/C single nucleotide polymorphisms, serum MIF concentration and genetic susceptibility to tuberculosis.Methods:1. Using case-control study, the test was divided into healthy control group, primary pulmonary tuberculosis group and secondary pulmonary tuberculosis group.2. Collect peripheral blood with EDTA anticoagulant and clinical information of specimens, then the blood serum was separated and stored at-80℃for testing. Extract genomic DNA from peripheral blood of all specimens.3. Detect MIF gene promoter-173G/C single nucleotide polymorphisms using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, Calculate genotype frequency and allele frequency using counting method and analyze the relationship between MIF-173G/C polymorphisms and tuberculosis susceptibility.4. Detect the MIF concentration of serum using ELISA method and compare its difference in different groups using SPSS17.0software.Result:1. Three genotypes were detected at-173G/C site of MIF. For MIF-173gene polymorphism, the frequency of GG,GC, and CC genotype in healthy control group, primary pulmonary tuberculosis group, and secondary pulmonary tuberculosis group was52.63%,47.37%,0,39.44%,57.75%、2.82%and56.58%、42.11%、 1.31%, respectively. The frequency of G allele and C allele in healthy control group, primary pulmonary tuberculosis group and secondary pulmonary tuberculosis group was76.32%.23.68%,68.31%、31.69%and77.63%.22.37%,respectively. Compare the frequency of GG, GC and CC genotype between health control group and primary pulmonary tuberculosis group,the frequency of GG genotype decreased and the frequency of GC, CC genotype increased in primary pulmonary tuberculosis group, the difference was significant (OR=1.706,95%C1:1.037-2.809,P<0.05); Compare the frequency of GG,GC and CC genotype between primary pulmonary tuberculosis group and secondary pulmonary tuberculosis group, the frequency of GG genotype increased and the frequency of GC, CC genotype decreased in primary pulmonary tuberculosis group, the difference was significant (OR=0.500,95%CI:0.284-0.879, P <0.05); Compare the frequency of GG, GC and CC genotype between health control group and secondary pulmonary tuberculosis group, the frequency of GG, CC genotype increased and the frequency of GC genotype decreased in secondary pulmonary tuberculosis group, the difference was significant(OR=0.853,95%CI:0.476-1.529, P>0.05). Compare the frequency of allele G and C between health control group and primary pulmonary tuberculosis group, the frequency of G allele decreased and the frequency of C allele increased in primary pulmonary tuberculosis group, the difference was significant (OR=1.495,95%CI:1.007-2.218,P<0.05). Compare the frequency of allele G and C between primary pulmonary tuberculosis group and secondary pulmonary tuberculosis group, the frequency of G allele increased and the frequency of C allele decreased in secondary pulmonary tuberculosis group, the difference was significant (OR=0.621,95%CI:0.394-0.980,P<0.05). Compare the frequency of allele G and C between health control group and secondary pulmonary tuberculosis group, the frequency of G allele increased and the frequency of C allele decreased in secondary pulmonary tuberculosis group, the difference wasn’t significant (OR=0.928,95%CI:0.570-1.513,P>0.05).2. MIF concentration of serum was5.750±3.260ng/ml in health control group, MIF concentration of serum was12.050±4.500ng/ml in primary pulmonary tuberculosis group and MIF concentration of serum was14.025±5.355ng/ml in secondary pulmonary tuberculosis group. MIF concentration of serum was5.460±4.442ng/ml in health control group with GG genotype, while it was5.880±1.115ng/ml in health control group with GC+CC genotype. MIF concentration of serum was12.290±15.100ng/ml in primary pulmonary tuberculosis group with GG genotype, while it was11.990±3.580ng/ml in primary pulmonary tuberculosis group with GC+CC genotype. MIF concentration of serum was15.350±6.450ng/ml in secondary pulmonary tuberculosis group with GG genotype,while it was13.130±6.190ng/ml in secondary pulmonary tuberculosis group with GC+CC genotype.Compared with MIF concentration in serum of health control group, MIF concentration increased in primary pulmonary tuberculosis group and secondary pulmonary tuberculosis group respectively, and the difference were all significant(P<0.05). Compared with MIF concentration in serum of primary pulmonary tuberculosis group, MIF concentration increased in secondary pulmonary tuberculosis group, but the difference wasn’t significant(P>0.05). Compared MIF concentration in serum of same group but different genotype, the difference were not significant in both health control group and primary pulmonary tuberculosis group between different genotypes(P>0.05). In secondary pulmonary tuberculosis group, compared with MIF concentration in serum of GC+CC genotype patients, MIF concentration increased in GG genotype patients, the difference was significant(P<0.05). Compared with MIF concentration in serum of health control group with GG genotype, MIF concentration increased in primary pulmonary tuberculosis group with GG genotype and secondary pulmonary tuberculosis group with GG genotype respectively, and the difference were both significant(P<0.05). Compared with MIF concentration in serum of health control group with GC+CC genotype, MIF concentration increased in primary pulmonary tuberculosis group with GC+CC genotype and secondary pulmonary tuberculosis group with GC+CC genotype respectively, and the difference were both significant(P<0.05). Compared with MIF concentration in serum of primary pulmonary tuberculosis group with GG genotype, MIF concentration increased in secondary pulmonary tuberculosis group with GG genotype, the difference was significant(P<0.05), Compared with MIF concentration in serum of primary pulmonary tuberculosis group with GC+CC genotype, MIF concentration increased in secondary pulmonary tuberculosis group with GC+CC genotype, but the difference wasn’t significant(P>0.05).Conclusion1. MIF gene promoter-173G/C site single nucleotide polymorphisms was related with TB susceptibility.Patients with GG genotype were not easy to get TB, but if they got TB, they were easy to relapse. Patients with GC+CC genotype were easy to get TB, but they were not easy to relapse.2. Compared with MIF concentration in serum of healthy people, MIF concentration increased significantly in TB patients. MIF concentration was higher in patients with GG genotype than patients with GG+GC genotype.3. MIF played an important role in the incidence and development of tuberculosis. |
PDF Full Text Request