The importance of rs1021737 and rs482843 polymorphisms of cystathionine gammalyase in the etiology of preeclampsia in the Caucasian population
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(19) 0 & #/ 1. Laboratory of Experimental Pharmacogenetics, Department of Clinical Pharmacy and Biopharmacy, University of Medical Sciences, Poznan, Poland Department of Pharmacology and Phytochemistry, Institute of Natural Fibers and Medicinal Plants, Poznan, Poland 3 Division of Perinatology and Women’s Diseases, Poznan University of Medical Sciences, Poznan, Poland 4 Laboratory of Molecular Biology in Division of Perinatology and Women’s Diseases, Poznan University of Medical Sciences, Poznan, Poland 5 Division of Gynecology and Obstetrics, Podhale Multidisciplinary Hospital, Nowy Targ, Poland 6 Department of General Pharmacology and Pharmacoeconomics, Pomeranian Medical University, Szczecin, Poland 2. Abstract Introduction: Recently, an increasing number of reports indicate the participation of genetic factors in the pathogenesis of preeclampsia (PE). The genes involved in the synthesis of nitric oxide that participates in the vasolidation, may play an important role in the development of this disorder. Hydrogen sulfide (H2S) which is produced by cystathionine gamma-lyase exhibits a similar effect to nitric oxide. It is suggested that certain polymorphisms of the CTH gene may participate in the development of chronic hypertension and preeclampsia. Aim of the study: To evaluate the frequency of genotypes and alleles of rs1021737 and rs482843 polymorphisms of CTH gene in women with preeclampsia from Wielkopolska region. Material and methods: The study group consisted of 60 patients with diagnosed preeclampsia, into the control group 120 healthy pregnant women were enrolled. The examined rs1021737 and rs482843 polymorphisms of CTH gene were determined using PCR-RFLP method. Results: Analysis of rs482843 polymorphism in the CTH gene showed a statistically significant difference in the prevalence of mutated GG genotype (p<0.000001) and mutated G allele (p<0.000001) in the group of pregnant women with PE compared to the control group. There was no such correlation for the rs1021737 polymorphism. Furthermore, there are also no relationship between studied polymorphisms and selected clinical and biochemical parameters.. Corresponding author: Agnieszka Seremak-Mrozikiewicz Division of Perinatology and Women’s Diseases Poznan University of Medical Sciences, Poznan, Poland 33 Polna Street, 60-535 Poznan e-mail: asm@data.pl. Nr 2/2015. © Polskie Towarzystwo Ginekologiczne. Otrzymano: 12.02.2014 Zaakceptowano do druku: 26.08.2014. 119.
(20) Ginekol Pol. 2015, 86, 119-125. P R A C E O R Y G I N A L N E poł ożn i ct wo. Przemysław M. Mrozikiewicz et al. The importance of rs1021737 and rs482843 polymorphisms of cystathionine gamma-lyase in the etiology of preeclampsia.... Conclusions: The results of rs482843 polymorphism analysis suggest that mutated GG genotype predisposes to preeclampsia occurrence. There was no such relationship for the rs1021737 polymorphism of CTH gene. Hence, further studies based on the determination of CSE expression level in women with PE may confirm the observed relationship between the rs482843 polymorphism and the risk of preeclampsia.. Słowa kluczowe: preeclampsia /
(21) / / / /. Streszczenie Wstęp: W ostatnim czasie coraz więcej doniesień wskazuje na udział czynników genetycznych w etiopatogenezie stanu przedrzucawkowego (PE – preeclampsia). Istotną rolę mogą odgrywać geny uczestniczące w syntezie tlenku azotu, głównego wazodylatora naczyń. Podobne działanie wazodyletacyjne pełni także siarkowodór (H2S) syntetyzowany przy udziale gamma-liazy cystationinowej. Sugeruje się, że określone polimorfizmy genu CTH mogą mieć znaczenie w rozwoju nadciśnienia przewlekłego i stanu przedrzucawkowego. Cel pracy: Określenie częstości występowania genotypów i alleli polimorfizmów rs1021737 i rs482843 genu CTH w grupie kobiet ze stanem przedrzucawkowym z regionu Wielkopolski. Materiał i metody: Grupa badana obejmowała 60 pacjentek ze zdiagnozowanym stanem przedrzucawkowym, natomiast grupa kontrolna stanowiła 120 zdrowych kobiet ciężarnych. Analizowane polimorfizmy rs1021737 i rs482843 genu CTH oznaczono przy pomocy metody PCR-RFLP. Wyniki: Analiza polimorfizmu rs482843 genu CTH wykazała różnicę istotną statystycznie w częstości występowania genotypu zmutowanego GG (p<0.000001) oraz zmutowanego allela G (p<0.000001) w grupie kobiet ciężarnych z PE w stosunku do grupy kontrolnej. Nie zaobserwowano takiej zależności dla polimorfizmu rs1021737. Stwierdzono również brak zależności między badanymi polimorfizmami a wybranymi parametrami klinicznymi i biochemicznymi. Wnioski: Uzyskane wyniki dotyczące polimorfizmu rs482843 genu CTH sugerują predyspozycję u ciężarnych nosicielek genotypu zmutowanego GG do wystąpienia stanu przedrzucawkowego. Nie wykazano takich uwarunkowań etiopatologicznych w odniesieniu do polimorfizmu rs1021737 genu CTH. Dalsze badania określające poziom ekspresji CSE u kobiet z PE mogą potwierdzić obserwowaną zależność pomiędzy polimorfizmem rs482843 a ryzykiem wystąpienia stanu przedrzucawkowego.. Słowa kluczowe: stan przedrzucawkowy / siarkowodór / / / /. Introduction 2
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(1007) 2!
(1008)
(1009) -
(1010)
(1011)
(1012) 2
(1013) -
(1014) !5
(1015) !
(1016) 2
(1017) 3+C&##+C
(1018) AN
(1019) pRBBBB4
(1020)
(1021) ? !
(1022) --
(1023) !5
(1024) 2!
(1025) !
(1026) - !
(1027) !5
(1028) 5!
(1029)
(1030) 6
(1031) !
(1032) !5
(1033) '
(1034) 2
(1035)
(1036) !
(1037) 3H+#OCCE/H
(1038) .
(1039) #/$//#O/B#
(1040)
(1041) pRBBBB4
(1042) 38
(1043) ;;4. Table I. Restriction enzymes and hydrolysis conditions.. Polymorphism. Restriction enzyme. Recognized sequence. Time/temperature incubation. Length of fragments (bp). rs482843. HindIII. . .
(1044) . 16 h, 37ºC. AA (172, 25) AG (197, 171, 25) GG (197). rs1021737. EcoRI. . .
(1045) . 16 h, 37ºC. GG (528) GT (528, 429, 99) TT (429, 99). Nr 2/2015. © Polskie Towarzystwo Ginekologiczne. 121.
(1046) Ginekol Pol. 2015, 86, 119-125. P R A C E O R Y G I N A L N E poł ożn i ct wo. Przemysław M. Mrozikiewicz et al. The importance of rs1021737 and rs482843 polymorphisms of cystathionine gamma-lyase in the etiology of preeclampsia.... Table II. Comparison of selected clinical parameters in preeclampsia and control groups. Study group (PE) n=60. Control group n=120. p. Age (years) mean±SD range median. 29.45±5.12 18-40 30. 28.74±4.75 18-42 29. ns. Gestational age at delivery (weeks) mean±SD range median. 33.57±3.82 25-40 33. 39.09±1.26 37-42 39. p<0.0001. Systolic blood pressure (mmHg) mean±SD range median. 160.92±20.22 110–220 160. 108.50±11.59 90–140 110. p<0.0001. Diastolic blood pressure (mmHg) mean±SD range median. 102.75±12.80 70–130 100. 67.63±9.12 50–90 70. p<0.0001. Height (cm) mean±SD range median. 164.93±6.43 149-176 165. 167.38±4.98 150–180 168. ns. Body weight before pregnancy (kg) mean±SD range median. 64.56±15.64 42-128 60. 60.33±9.38 40–97 59.5. p<0.009. 80.59±15.83 55-139 76. 75.34±10.52 52–106 59.5. Infant birth weight (g) mean±SD range median. 1963±887.49 620-3940 1850. 3454.43±403.12 2560-4640 3430. p<0.0001. Number of pregnancies mean±SD range median. 1.53±0.83 1–4 1. 1.55±0.79 1–6 1. ns. Delivery mode (n) Spontaneous birth Cesarean section. 7 (11.67%) 53 (88.33%). 56 (46,67%) 64 (53.33%). p<0.0001. Hemoglobin (mmol/l) mean±SD range median. 8.46±2.54 0.90–15.30 7.80. 6.72±0.79 4.5–8.8 6.775. p<0.0001. Erythrocytes (10^6/μl) mean±SD range median. 4.28±0.95 3.24–9.92 4.08. 3.68±0.45 2.5–4.81 3.685. p<0.0001. Leukocytes (10^3/μl) mean±SD range median. 10.85±4.07 4.14–25.29 9.90. 23.39±72.20 8.27–806 16.23. ns. Platelets (g/l) mean±SD range median. 207.95±66.69 88.00–398.00 210.50. 199.77±62.84 9.1–314.6 206.45. ns. Hematocrit (l/l) mean±SD range median. 0.36±0.04 0.20–0.45 0.36. 0.33±3.26 0.219–0.36 0.32. ns. Protein in urine (mg/dl) mean±SD range median. 357.74±253.11 166.8–1336.8 500. 10.82±12.44 0-25 0. p<0.0001. Parameter. Body weight during pregnancy (kg) mean±SD range median. 122. © Polskie Towarzystwo Ginekologiczne. p<0.026. Nr 2/2015.
(1047) Ginekol Pol. 2015, 86, 119-125. P R A C E. O R Y G I N A L N E po ł o ż n i c t wo. Przemysław M. Mrozikiewicz et al. The importance of rs1021737 and rs482843 polymorphisms of cystathionine gamma-lyase in the etiology of preeclampsia.... Table III. Frequency of genotypes of rs1021737 polymorphism of the CTH gene in women with preeclampsia and the control group. Study group (PE). Control group. Observed n (%). Expected %. Observed n (%). Expected %. GG. 27 (45.00). 46.69. 52 (43.33). 45.00. GT. 28 (46.67). 43.28. 57 (47.50). 44.16. TT. 5 (8.33). 10.03. 11 (9.17). 10.84. 60 (100.00). 100.00. 120 (100.00). 100.00. Total G. 161 (67.08). 82 (68.33). T. 79 (32.92). 38 (31.67). 240 (100.00). 120 (100.00). Total. Table IV. Comparison of the odds ratio (OR) and 95% confidence interval (CI) for each genotype of the rs1021737 polymorphism. Polymorphism rs1021737. OR. 95% CI. p. GG. 1.07. 0.55 – 2.01. 0.48. GT. 0.97. 0.49 – 1.88. 0.52. TT. 0.90. 0.23 – 2.98. 0.55. G. 1.06. 0.65 – 1.75. 0.45. T. 0.94. 0.57 – 1.55. 0.45. Table V. Frequency of genotypes of rs482843 polymorphism of the CTH gene in women with preeclampsia and the control group. Study group (PE). Control group. Observed n (%). Expected %. Observed n (%). Expected %. AA. 16 (26.67). 19.51. 66 (55.00). 57.50. AG. 21 (35.00). 49.32. 50 (41.67). 36.66. GG Total. 23 (38.33). 31.17. 4 (3.33). 5.84. 60 (100.0). 100.00. 120 (100.00). 100.00. A. 53 (44.17). G. 67 (55.83). 58 (24.17). 120 (100.00). 240 (100.00). Total. 182 (75.83). Table VI. Comparison of the odds ratio (OR) and 95% confidence (CI) interval for each genotype of the rs482843 polymorphism. Polymorphism rs482843. OR. 95% CI. p. AA. 0.29. 0.14 – 0.61. 0.00025. AG. 0.75. 0.37 – 1.49. 0.24. GG. 18.03. 5.56 – 74.89. <0.000001. A. 0.25. 0.15 – 0.41. <0.000001. G. 3.97. 2.42 – 6.49. <0.000001.
(1048) -
(1049) !5
(1050)
(1051) 225
(1052) -
(1053) !5
(1054)
(1055)
(1056) 6
(1057)
(1058) ! !!
(1059) ? !
(1060) --
(1061)
(1062) !5
(1063) !
(1064) 2
(1065)
(1066) !
(1067) !
(1068) !5
(1069) -J
(1070) -
(1071) !2
(1072) 3GG
(1073) 5!
(1074) pVB/C
(1075) GT
(1076) 5!!
(1077) pVB$
(1078) ! !
(1079) 5
(1080) TT !2
(1081) pVB$$4
(1082) ;
(1083) !
(1084)
(1085)
(1086) -J
(1087) -
(1088) !5
(1089) G
(1090)
(1091).
(1092) !5
(1093) ! !
(1094) T
(1095)
(1096) -
(1097) !5
(1098)
(1099) 225
(1100)
(1101) 6
(1102)
(1103)
(1104) !5
(1105) '
(1106)
(1107) !
(1108) 38
(1109) ;;;
(1110)
(1111) ;<4. Nr 2/2015. ;6
(1112) !2
(1113) -
(1114) !5
(1115)
(1116) 225
(1117) -
(1118) !5
(1119)
(1120)
(1121)
(1122)
(1123) !5
(1124) '
(1125)
(1126) !5
(1127) !
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