Background
Methods
Subjects and phenotypical characteristics
Asthma (n = 467) | Control (n = 288) | |
---|---|---|
Mean age, years | 40.8 ± 14.1 | 45.1 ± 0.7 |
Range | 18-76 | 22-72 |
Sex, n (%) | ||
Male | 185 (39.6) | 158 (55.0) |
Female | 282 (60.4) | 130 (45.0) |
logeIgE, U/L | 5.24 ± 1.44 | 3.64 ± 1.10 |
Eosinophils,% | 1.63 ± 0.91 | 2.19 ± 0.93 |
Lung function | ||
FEV1,% | 68.94 ± 37.88 | 90.6 ± 14.92 |
FEV1/FVC,% | 63.93 ± 14.91 | 81.53 ± 9.52 |
SNPs selection and genotyping
Gene | Region | PCR primers(5'→3') | Sequencing primer(5'→3') | Annealing temperature(°C) | PCR product (bp) | Restriction Enzyme | RFLP products (bp) |
---|---|---|---|---|---|---|---|
VDR
| |||||||
Exon 1 | F:TGGCACCAAGGATGCCAGCT | 57 | 364 | ||||
R:TCCTGCTCCTGTGGCTGTGA | R:GTGAGCGCCGCATGTTCC | ||||||
Exon 2 | F:CCTCATGTCTTCTGTTGGAG | F:CCTCATGTCTTCTGTTGGAG | 57 | 438 | |||
R:TGCATCTGACCCTGGACTTC | |||||||
Exons 3,4 | F: CCTATCTTGGACCTTTACCC | F: CCTATCTTGGACCTTTACCC | 57 | 758 | |||
R:TCCATTAGGGAGCCTTCCAC | |||||||
Exon 5 | F: CTGTGGAGTCACTGTGGGAT | F: CTGTGGAGTCACTGTGGGAT | 57 | 390 | |||
R:GAAGTGGTGGATGAGTGATC | |||||||
Exons 6,7 | F: TGTTGGTGCCCAGCAGGTGT | F: TGTTGGTGCCCAGCAGGTGT | 57 | 680 | |||
R:GCTACGTCTCCCTTCAGGTT | |||||||
Exon 8 | F: TGCTGCCGTTGAGTGTCTGT | 57 | 526 | ||||
R:GGTTGGACAGGAGAGAGAAT | R:ACAGGAGAGAGAATGGGCTG | ||||||
CYP2R1
| |||||||
Exon 1 | F:GGGACCTGAGGGTATCCTGC | F:ACCTGAGGGTATCCTGCCAATG | 55 | 409 | |||
R:GGCGGCCATAAGTCCAACC | |||||||
Exon 2 | F: AGGGACGTAACAAATCCTGG | F: AGAGCCGCACTTACCACTTG | 57 | 412 | |||
R: GTAGTACAGCCTGAAAGGTC | |||||||
Exon 3 | F: AACAGGACCCAACCATGTAG | F: GGAGGACAATTTGGAGAAGG | 57 | 881 | |||
R: CATCGCAGGAGTTCCTAAAG | |||||||
Exon 4 | F: TGGCTTGTTATAGGTTATCC | 57 | 574 | ||||
R: CATTCTCTCCTGTTAGAATC | R: CATTCTCTCCTGTTAGAATC | ||||||
Exon 5 | F: GTTCTGCTTGCTGAAGTGTC | F: CATAGTTCCCTCTTTCTTTG | 57 | 415 | |||
R: ACCAAGTTCAGGGATAAGGC | |||||||
GC
| |||||||
rs4588 rs7041 | F:AAATAATGAGCAAATGAAAGAAGAC | 55 | 483 | Hae III | 297/186 | ||
R:CAATAACAGCAAAGAAATGAGTAGA | Sty I | 305/178 |
Haelll/Styl Genotypes | Potential haplotypes | Deduced haplotypes | Corresponding DBP genotypes | Asthma (N) | Control (N) |
---|---|---|---|---|---|
HH SS | HS/HS | HS/HS | Gc1F/Gc1F | 72 | 41 |
HH Ss | HS/Hs | HS/Hs | Gc1F/Gc2 | 129 | 59 |
HH ss | Hs/Hs | Hs/Hs | Gc2/Gc2 | 24 | 1 |
Hh SS | HS/hS | HS/hS | Gc1F/Gc1S | 93 | 46 |
Hh Ss | Hs/hS or HS/hs* | Hs/hS | Gc2/Gc1S | 71 | 35 |
hh/SS | hS/hS | hS/hS | Gc1S/Gc1S | 27 | 18 |
Hh/Ss | hS/hs* | Gc1s/not deduced | Not assigned | 0 | 0 |
Statistical analysis
Results
Genotype/Allele | Asthmatics, n (%) | Controls, n (%) | χ2
|
P
| |
---|---|---|---|---|---|
VDR rs2228570 | |||||
CC | 152 | 60 | |||
CT | 230 | 91 | |||
TT | 85 | 37 | 0.194 | 0.908 | |
C | 534 (57.2) | 211 (56.1) | |||
T | 400 (42.8) | 165 (43.9) | 0.122 | 0.727 | |
CYP2R1 rs12794717 | |||||
CC | 171 | 64 | |||
CT | 214 | 86 | |||
TT | 81 | 25 | 1.02 | 0.601 | |
C | 556 (59.7) | 214 (61.1) | |||
T | 376 (40.3) | 136 (38.9) | 0.234 | 0.628 | |
GC
| |||||
Gc1/1 | 192 | 105 | |||
Gc1/2 | 200 | 94 | |||
Gc2/2 | 24 | 1 | 10.401 |
0.006
| |
Gc1 | 584 (70.2) | 304 (76.0) | |||
Gc2 | 248 (29.8) | 96 (24.0) | 4.527 |
0.033
|
Serum 25OHD | Subjects, n | Genotypes, n (%) |
P value | ||
---|---|---|---|---|---|
CYP2R1 rs12794717 | |||||
CC
|
CT
|
TT
| |||
≥50 nmo l/L | 49 | 20 (40.8) | 24 (49) | 5 (10.2) | |
< 50 nmol/L | 363 | 132(36.4) | 167 (46) | 64 (17.6) | 0.42 |
VDR rs2228570 | |||||
CC | CT | TT | |||
≥50 nmol/L | 49 | 13 (26.5) | 25 (51.1) | 11 (22.4) | |
< 50 nmol/L | 362 | 124 (34.3) | 180 (49.7) | 58 (16) | 0.397 |
GC rs4588 and rs7041 | |||||
Gc1/1 | Gc1/2 | Gc2/2 | |||
≥50 nmol/L | 49 | 20 (40.8) | 26 (53.1) | 3 (6.1) | |
< 50 nmol/L | 363 | 169 (46.5) | 173 (47.7) | 21 (5.8) | 0.747 |