Effect of Metoclopramide on G6PDH Activity in First Trimester of Pregnancy
Huda Shaker Arif 1*, Sameaa J. Khammas 2 Mohammed I. Hamzah3
Department of chemistry, College of Science women; University of Baghdad, Baghdad, Iraq
Corresponding Author Email *: [email protected]
ABSTRACT
The aim of this study was to investigate effect of Metoclopramide drugs on G6PD and antioxidant enzymes. Metoclopramide drugs are currently being used to Reduce or prevent nausea and vomiting in patients. This is the first study to show effect of Metoclopramide on glucose-6-phosphate dehydrogenase (G6PD) and antioxidant enzyme activities in first trimester in pregnancy. For in vitro studies, G6PD was purified from human erythrocyte Metoclopramide hydrochloride showed no inhibition effects
Keywords: Glucose-6-phosphate dehydrogenase; Antioxidant enzymes; Inhibition, Metoclobramide drug,
Introduction
Metoclopramide hydrochloride (Figure. 1) is Benzamide, 4-amino-5-chloro-N-[2-(diethylamino) ethyl]-2- methoxy-, mono hydrochloride, monohydrate; 4-Amino5-chloro -N-[2-(diethylamino) ethyl]-o-anisamide mono hydrochloride monohydrate. [1] Symptomatic treatment of nausea and vomiting including that associated with acute migraine Delayed (but not acute) chemotherapy- induced nausea and vomiting Radiotherapy-induced nausea and vomiting Prevention of postoperative nausea and vomiting. [2,3].
Glucose-6-phosphate dehydrogenase (d-glucose-6-phos phate: NADP+ oxidoreductase EC 1.1.1.49; G6PD) oxidizes glucose-6-phosphate to 6-phosphogluconolactone, reducing NADP+ to NADPH. The pentose phosphate pathway (PPP) is the only source of NADPH in the erythrocytes. The main function of the pathway seems to be to protect the erythro cytes against oxidative damage that is caused by free radical in the number of molecules in cells, including membrane lipids, proteins, and nucleic acids [4,5]. Formation of these harmful radicals is an occurring intracellular metabolic pro cess [6,7].
Aim of Study
1- To estimate G6PDH levels in pregnant women with Metoclopramide and control group.
2- To find any correlation between G6PDH and other external factor and some BMI parameter in all study group
Methodology
In this study, taken Forty blood samples in the EDTA tube for pregnant women in the first trimester of pregnancy, as they were collected from (Imamine Al-Kadhimin Medical City Hospital) for forty orders, after which g6pdh was measured in the laboratories of the Nahrain Medical College, after which the statistical work was done with SPS.
The enzyme activity is determined by measurement of the rate absorbance change at 340 nm due to the reduction of NADP. G-6-P
G-6-P + NADP G-6-PDH gluconate-6-p + NADPH + H+
SAMPLE:
Erythrocytes.
PREPARATION OF SAMPLE:
Wash 0.2 ml of blood with 2 ml aliquots of 0.9% NaCl solution. Centrifuge after each wash for 10 min at around 3000 rpm. Repeat 3 times. Suspend the washed centrifuged erythrocytes in 0.5 ml of solution 4 and let stand for 15 min at +4°C and then centrifuge again.
Use the supernatant in the assay within 2 hours. [8-10]
Device name: Apel PD-303 spectrophotometer
MANUAL USE
Wavelength: 340 nm (Hg 334 nm or Hg 365 nm) Cuvette: I cm light path
Temperature: +37°C Measurement: against air Pipette into test tube
Semi Micro
RI 1.00 ml
R2 0.003 ml
Haemolysate 0.015 ml
Mix .incubate for 5 minutes at+37°C , then add , R3 0.015 ml
Mix ,read initial absorbance and start timer simultaneously Read again after 1,2 and 3 minutes
The following equation is used .
G-6-PDH mU/gHb = mU .erthrocytes per ml x 100
Hb (g/di ) x 1000
100 =Factor to convert from ml to di
Hb(g/di) = Haemoglobin concentration determined for each specimen
RESULTS
The total number of study participants was 40. All of them were pregnant women in the 1st trimester and they were divided into two groups: Metoclopramide group included 20 pregnant women used metoclopramide for treatment of nausea and vomiting and control group included 20 pregnant women didn’t use metoclopramide.
3.1. Age
The distribution of study groups by age is shown in figure (3.1). Study participants’ age was ranging from 20 to 31 years with a mean of 24.6 years and a standard deviation (SD) of ± 2.9 years. The highest proportion of study participants in metoclopramide group was aged < 25 years (55%); while 60% of controls were aged ≥ 25 years (60%).
Figure 3.1: Distribution of study groups by age METOCLOPRAMIDE CONTROL
55% 40% 45%
60%
Age (Year)
< 25
≥ 25
3.2. Body Mass Index (BMI)
Figure 3.2 shows the distribution of study groups by BMI level. In this study, the highest proportion of study participants in metoclopramide and control groups was overweighed (55%
and 45% respectively).
Figure 3.2: Distribution of study groups by BMI level
The comparison in age and BMI between study groups is shown in table (3.1). No statistical significant differences (P ≥ 0.05) in age and BMI between study groups.
Table 3.1: Comparison in certain information between study groups Variable
Study group
P - Value Metoclopramide
Mean ± SD
Control Mean ± SD
Age (Year) 24.6 ± 3.0 24.5 ± 3.0 0.988
BMI (kg/m2) 28.57 ± 3.0 27.65 ± 2.8 0.324
3.3. Hemoglobin and PCV
The comparison in Hb and PCV between study groups is shown in table (3.2). Means of Hb and PCV were significantly higher in controls than that in metoclopramide group (12.7 versus 12.1 g/dl, P= 0.003; and 37.5 versus 35.6%, P= 0.006 respectively).
15%
25%
55%
45%
30% 30%
Metoclopramide Control
BMI Level
Normal Overweight Obese
Table 3.2: Comparison in hemoglobin and PCV between study groups Variable
Study group
P - Value Metoclopramide
Mean ± SD
Control Mean ± SD
Hemoglobin (g/dl) 12.1 ± 0.6 12.7 ± 0.6 0.003
PCV (%) 35.6 ± 2.2 37.5 ± 1.9 0.006
3.4. G6PD
The comparison in G6PD level between study groups is shown in figure and table (3.3).
We noticed that there was no statistical significant difference in G6PD level between study groups (5.35 versus 5.34 U/g, P= 0.959).
Figure 3.3: G6PD level in study groups
Table 3.3: Comparison in G6PD level between study groups
G6PD level (U/g)
Study group
P - Value Metoclopramide
Mean ± SD
Control Mean ± SD
5.35 ± 0.62 5.34 ± 0.59 0.959 Correlation between G6PD level with age and BMI
Weak statistically significant association detected between G6PD level and BMI (r=
0.377, P= 0.005). No statistically significant correlation was detected (P = 0.189) between G6PD level and age as shown in table (3.4).
5.35 5.34
G6PD level (U/g)
Metoclopramide Control
Table 3.4: Correlation between G6PD level with age and BMI
Variable G6PD level (U/g)
r P - Value
Age (Year) 0.212 0.189
BMI (kg/m2) 0.377 0.005
Statistical analysis
The data analyzed using Statistical Package for Social Sciences (SPSS) version 26. The data presented as mean, standard deviation and ranges. Categorical data presented by frequencies and percentages. Independent t-test (two tailed) was used to compare the continuous variables accordingly. Pearson’s correlation test (r) was used to assess correlation between G6PD level and both of age and BMI. A level of P – value less than 0.05 was considered significant.
REFRENCES
[1] The United State Pharmacopeia Convection, Inc, 41- NF, 2018; 36: 2699.
[2] British National Formulary (BNF) Royal Pharmaceutical Society, 2016; (70): 347.
[3] Sweetman S. Martindale: The Complete Drug Reference. The Pharmaceutical Press: London, 2009; 36: 502.
[4] Mates JM, Perez-Gomez C, Nunez De Castro I. Antioxidant enzymes and human diseases.
Clin Biochem 1999;32(8):595–603.
[5] ÖZMEN, İ., Ciftci, M., KÜFREVİOĞLU, Ö. İ., & ÇÜRÜK, M. A. (2004). Investigation of glucose 6-phosphate dehydrogenase (G6PD) kinetics for normal and G6PD-deficient persons and the effects of some drugs. Journal of enzyme inhibition and medicinal chemistry, 19(1), 45-50.
[6] Ames BN, Shigenaga MK, Hagen TM. Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci USA 1993;90:7915–22.
[7] McCord JM. Human disease, free radicals, and the oxi dant/antioxidant balance. Clin Biochem 1993; 26:351–7
[8]. Kornberg, A. et al, Methods in Enzymology I, Academic Press, New York, 1955; p.323.
[9]. Makarem, A Clinical Chemistry-Principles and Techniques. 2d Ed. R.F. Henry, D. C.
Cannon, J.W. Winkelman, Editors. Harper and Row, Hagerstown [MD]. 1974; I128-1135.
[10]. Lohr GW, Waller HD: Glucose-6-Phosphate Dehydrogenase. Methods of Enzymatic Analysis, 3rd Edition - Varlag Chemie, Wehnheim: 1974; p. 636
[11]. Yogesh Hole et al 2019 J. Phys.: Conf. Ser. 1362 012121
