Analysis of Gln223Agr Polymorphism of Leptin Receptor Gene in Type II Diabetic Mellitus Subjects among Malaysians
Abstract
:1. Introduction
2. Results and Discussion
2.1. Genotypic and Allelic Frequency
3. Methodology
3.1. Study Population
3.2. Sampling Method
3.3. Genotyping
3.4. Enzymatic Digestion
3.5. DNA Sequencing
3.6. Data Validation
3.7. Statistical Methods
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Characteristics | T2DM | Lower/Upper | Control | Lower/Upper | p-value | ||||
---|---|---|---|---|---|---|---|---|---|
Male | Female | Total | CI 95% | Male | Female | Total | CI 95% | ||
Age (years) | 60.7 ± 10.2 | 61.4 ± 8.8 | 61 ± 9.8 | 59.9–62.2 | 56.1 ± 11.2 | 49.7 ± 13 | 53.3 ± 12.4 | 51.8–54.8 | 0.000 ** |
BMI (Kg/m2) | 27.47 ± 4.38 | 28.29 ± 5.66 | 27.73 ± 4.83 | 27.1–28.3 | 26.42 ± 4.32 | 26.07 ± 4.94 | 26.27 ± 4.59 | 26.8–26.1 | 0.014 * |
WHR | 0.95 ± 0.05 | 0.93 ± 0.07 | 0.94 ± 0.06 | 0.94–0.95 | 0.92 ± 0.05 | 0.86 ± 0.07 | 0.9 ± 0.06 | 0.89–0.91 | 0.000 ** |
SBP (mm Hg) | 140.59 ± 22.55 | 144.37 ± 22.63 | 141.83 ± 22.6 | 139.1–144.56 | 138.34 ± 20.69 | 134.21 ± 18.75 | 136.64 ± 19.98 | 134.24–139.05 | 0.048 * |
DBP (mm Hg) | 80.05 ± 9.89 | 78.25 ± 9.79 | 79.45 ± 9.88 | 78.26–80.65 | 79.47 ± 9.47 | 78.03 ± 9.4 | 78.87 ± 9.45 | 77.73–80.02 | 0.170 |
FPG (mmol/L) | 8.29 ± 3.17 | 8.67 ± 3.4 | 8.41 ± 3.24 | 8.02–8.81 | 5.49 ± 0.98 | 4.79 ± 0.81 | 5.19 ± 0.97 | 5.07–5.31 | 0.000 ** |
HbA1C | 8.22 ± 1.94 | 8.27 ± 1.64 | 8.24 ± 1.84 | 8.02–8.45 | 5.85 ± 0.51 | 5.7 ± 0.38 | 5.79 ± 0.46 | 5.73–5.86 | 0.000 ** |
LDL (mmol/L) | 2.31 ± 0.78 | 2.46 ± 0.89 | 2.36 ± 0.82 | 2.26–2.46 | 2.74 ± 0.97 | 2.71 ± 0.97 | 2.73 ± 0.97 | 2.61–2.84 | 0.060 |
HDL (mmol/L) | 1.11 ± 0.28 | 1.31 ± 0.36 | 1.17 ± 0.32 | 1.13–121 | 1.16 ± 0.33 | 1.34 ± 0.42 | 1.23 ± 0.38 | 1.19–1.28 | 0.000 ** |
TG (mmol/L) | 1.61 ± 0.81 | 1.62 ± 0.75 | 1.61 ± 0.79 | 1.52–1.71 | 1.5 ± 0.83 | 1.04 ± 0.49 | 1.3 ± 0.74 | 1.28–1.38 | 0.001 ** |
Chol (mmol/L) | 4.17 ± 0.96 | 4.59 ± 1.04 | 4.3 ± 1.01 | 4.18–4.43 | 4.56 ± 1.03 | 4.67 ± 1.25 | 4.61 ± 1.13 | 4.47–4.74 | 0.003 ** |
CVD Risk% | 3.97 ± 1.25 | 3.78 ± 1.31 | 3.9 ± 1.27 | 3.75–4.06 | 4.15 ± 1.28 | 3.67 ± 1.17 | 3.94 ± 1.26 | 3.79–4.09 | 0.014 ** |
Family History | |||||||||
No | 97 (51.1) | 45 (49.5) | 142 (50.5) | 104 (66.2) | 69 (56.1) | 173 (61.8) | 0.007 **,† | ||
Yes | 93 (48.9) | 46 (50.5) | 139 (49.5) | 53 (38.8) | 54 (43.9) | 107 (38.2) | |||
Duration (years) | 10 ± 8 | 10.6 ± 8.2 | 10.2 ± 8.1 | - | - | - | - |
T2DM (n-284) | Controls (n-281) | ||||||
---|---|---|---|---|---|---|---|
Genotypes and Alleles | Malay (n = 145) | Chinese (n = 49) | Indians (n = 90) | Malay (n = 133) | Chinese (n = 71) | Indians (n = 77) | p value |
WT (GG) | 28.7 | 26.1 | 41.2 | 16.4 | 8.8 | 30.1 | 0.057 |
HOM (AA) | 59.6 | 73.9 | 50.6 | 68.8 | 83.8 | 50.7 | 0.011 * |
HET (GA) | 11.8 | - | 8.2 | 14.8 | 7.4 | 19.2 | 0.095 |
Alleles | |||||||
G | 0.346 | 0.261 | 0.453 | 0.238 | 0.125 | 0.397 | NS |
A | 0.654 | 0.739 | 0.547 | 0.762 | 0.875 | 0.603 | |
Post-Hoc Test | p value | (95% Confidence interval(lower bound-upper bound) | |||||
Chinese alone | |||||||
GG vs. AA | 0.019 * | 0.50–0.54 | |||||
GG vs. GA | 0.007 * | 0.19–1.14 | |||||
AA vs. GA | 0.091 | −0.05–0.81 | |||||
Three Ethnics | |||||||
GG vs. AA | 0.001 * | 0.80–0.27 | |||||
GG vs. GA | 0.001 * | 0.11–0.41 | |||||
AA vs. GA | 0.001 * | −0.41–−0.11 |
Factors | T2DM | Control | p-value | ||||
---|---|---|---|---|---|---|---|
WT (GG) | HOM (AA) | HET (GA) | WT (GG) | HOM (AA) | HET (GA) | ||
Age (years) | 61.8 ± 9.8 | 60.5 ± 9.5 | 60.2 ± 9.9 | 56.4 ± 11.2 | 53.8 ± 12.0 | 52.3 ± 14.9 | 0.007 ** |
BMI (Kg/cm2) | 27.87 ± 5.26 | 27.67 ± 4.73 | 27.4 ± 4.31 | 26.43 ± 4.67 | 26.31 ± 4.67 | 26.35 ± 4.13 | 0.423 |
WHR | 0.94 ± 0.06 | 0.94 ± 0.06 | 0.93 ± 0.03 | 0.91 ± 0.05 | 0.89 ± 0.06 | 0.9 ± 0.1 | 0.022 ** |
SBP (mm Hg) | 145.6 ± 19.9 | 139.2 ± 23.1 | 144.3 ± 22.7 | 135.36 ± 18.65 | 137.8 ± 20.4 | 134.5 ± 19.6 | 0.334 |
DBP (mm Hg) | 80.27 ± 8.7 | 78.97 ± 10.1 | 79.34 ± 7.8 | 77.48 ± 8.17 | 79.32 ± 9.82 | 78.52 ± 9.16 | 0.385 |
FPG (mmol/L) | 8.28 ± 3.2 | 8.55 ± 3.27 | 7.58 ± 3.1 | 5.59 ± 0.75 | 5.17 ± 0.99 | 5.03 ± 0.91 | 0.000 ** |
HbA1C | 8.13 ± 1.7 | 8.26 ± 1.8 | 8.43 ± 2.2 | 5.83 ± 0.46 | 5.79 ± 0.47 | 5.76 ± 0.54 | 0.000 ** |
LDL (mmol/L) | 2.41 ± 0.88 | 2.32 ± 0.74 | 2.55 ± 1.22 | 2.57 ± 0.9 | 2.82 ± 0.97 | 2.59 ± 1.09 | 0.000 ** |
HDL (mmol/L) | 1.16 ± 0.25 | 1.17 ± 0.35 | 1.12 ± 0.29 | 1.25 ± 0.33 | 1.24 ± 0.38 | 1.22 ± 0.42 | 0.003 ** |
TG (mmol/L) | 1.54 ± 0.61 | 1.65 ± 0.88 | 1.36 ± 0.64 | 1.43 ± 0.71 | 1.31 ± 0.78 | 1.14 ± 0.54 | 0.139 |
Chol (mmol/L) | 4.44 ± 1.13 | 4.25 ± 0.92 | 4.18 ± 1.15 | 4.47 ± 0.97 | 4.7 ± 1.15 | 4.43 ± 1.28 | 0.010 ** |
CVD Risk% | 3.99 ± 1.23 | 3.9 ± 1.29 | 3.67 ± 0.89 | 3.79 ± 1.06 | 3.98 ± 1.28 | 3.82 ± 1.2 | 0.216 |
Family History | |||||||
No | 37 (43.5) | 86 (54.8) | 11 (50) | 31 (63.3) | 108 (59.3) | 27 (71.1) | 0.000 **,† |
Yes | 48 (56.5) | 71 (45.2) | 11 (50) | 18 (36.7) | 74 (40.7) | 11 (28.9) | |
Duration (years) | 10.9 ± 8.4 | 9.9 ± 7.9 | 8.6 ± 6.3 | - |
SNP ID | Amino acid changing region | Enzymatic digestion time temperature and volume | Cutting position | Forward and reverse primer | PCR product size (bp) | PCR cycling condition (time and temperature) | Reference |
---|---|---|---|---|---|---|---|
rs1137101 | Gln223Arg | Msp I 45 min 37 °C 4 Unit | 5′...C^C G G...3′ 3′...G G C^C...5′ | 5′-CAAACTCAACGACACTCTCCTT-3′ 5′-CTGAACTGACATTAGAGGTGAC-3′ | 80 | 5 min 95 °C 45 s 95 °C 30 s 57 °C 45 s 72 °C 30 cycle 10 min 72 °C 4 °C ∞ | [37] |
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Etemad, A.; Ramachandran, V.; Pishva, S.R.; Heidari, F.; Aziz, A.F.A.; Yusof, A.K.M.; Pei, C.P.; Ismail, P. Analysis of Gln223Agr Polymorphism of Leptin Receptor Gene in Type II Diabetic Mellitus Subjects among Malaysians. Int. J. Mol. Sci. 2013, 14, 19230-19244. https://doi.org/10.3390/ijms140919230
Etemad A, Ramachandran V, Pishva SR, Heidari F, Aziz AFA, Yusof AKM, Pei CP, Ismail P. Analysis of Gln223Agr Polymorphism of Leptin Receptor Gene in Type II Diabetic Mellitus Subjects among Malaysians. International Journal of Molecular Sciences. 2013; 14(9):19230-19244. https://doi.org/10.3390/ijms140919230
Chicago/Turabian StyleEtemad, Ali, Vasudevan Ramachandran, Seyyed Reza Pishva, Farzad Heidari, Ahmad Fazli Abdul Aziz, Ahmad Khairuddin Mohamed Yusof, Chong Pei Pei, and Patimah Ismail. 2013. "Analysis of Gln223Agr Polymorphism of Leptin Receptor Gene in Type II Diabetic Mellitus Subjects among Malaysians" International Journal of Molecular Sciences 14, no. 9: 19230-19244. https://doi.org/10.3390/ijms140919230