1. INTRODUCTION:

Polycystic ovary syndrome (PCOS):

Is the most frequent endocrine disorder in women of reproductive age with heterogeneous clinical and biochemical features.¹ The diverse and complex female endocrine disorder PCOS affects 1 in 15 women worldwide.² PCOS produces symptoms in approximately 5% to 10% of women of reproductiveage 12–45 years old.³The major characteristics of the syndrome are chronic anovulation, menstrual abnormalities (oligomenorrhea or amenorrhea), hyper androgenism, and PCOS appearance using ultrasound.⁴ PCOS is associated with obesity, Insulin resistance, and increased risk of type 2 diabetes mellitus,dyslipidemia, cardiovascular disease, and endometrial carcinoma,⁵and elevated testosteronelevels. Research suggests that anxiety, depression, and various other psychological problems are greater in PCOS than in healthy women.⁶

Nickel was measured by atomic absorption spectroscopy, while lead wasdeterminedaccording to the literature method.^7-9

2. MATERIALS AND METHODS:

The study was conducted with 70 women with PCOS disease and 50 control cases.The samples were collected from infertility unit at Al Hussein Teaching Hospital in Thi-Qar/Iraq. PCOS was defined when at least two of the following three features were present: oligo-/amenorrhea, Hyperandrogenism (and/or hirsutism), and polycystic ovaries. To compared with control group, we randomly selected 50 samples. None of these had symptoms of Hyperandrogenism, menstrual dysfunction, and infertility. Nitric acid, perchloric acid, and hydrochloric acid were used in the digestion processes.

2.1 Laboratory Analysis:

All cases were studied within the first 1-5 days after onset of menstruation. A sample of blood (5ml) was centrifuged (2500 rpm) for 10 minutes to get serum. The serum was kept at -20ᵒC. The metal concentrations were determined by Atomic absorption spectrophotometric.^10,11

2.2 Digestion of serum:

The digestion of serum was achieved by adding nitric acid (2ml, 70%) and perchloric acid (1ml, 70%) to the serum (0.5 ml) in a test tube. After heating the solution for 1 hour at 160ºC in oil bath, the solution was cooled. The volume was completed to 10 ml by adding hydrochloric acid (0.3N).¹²

2.3 Statistical Analysis:

The statistical analysis was achievedby using Microsoft Excel 2010 the results were expressed as mean ± standard deviation (mean ± SD). One-way ANOVA was used to compare parameters in different studied groups. Pearson's correlation was applied to determine the relationship between the present study parameters. P-values (P≤0.05) were considered statistically significant.

3. RESULT AND DISCUSSION:

3.1 The levels of trace elements (Ni, Pb, Fe, and Zn) in patients (PCOS) and control group with age general from 10-40 years are discussed in this section. Note each value represents mean±SD values with non-identical superscript (a and b) for vertical comparison are considered significantly differences (p≤0.05).

3.1.1 Nickel:

The present study showed that Ni levelswere significantly higher in PCO patients thanin control groups (p≤0.05).The results were in consistent with literature.⁹Nickel levels are higher in PCOS in comparison with control group.suggesting that nickel affects PCOS by increasing reproductive hormone levels related with oxidative stress. It seems that nickel disrupts physiological homeostasis, inducing glucose deregulation through ROS pathways¹⁸.

Table 1 The nickel concentrationsin serum (ppm).

Group	No.	Ni Mean ±SD
Patient	70	0.69±0.19 a
Control	50	0.33±0.09 b
LSD		0.04

LSD:Least Significant Difference*

Figure 1The nickel levels in patient and control groups.

3.1.2 Lead:

The resultsshowed that the concentration of lead was significantly higher in PCO patients than in control groups (p≤0.05), may be due to increasingthe accumulation in body with age and increasing FSH and LH hormone concentrations.¹³

Table 2: The concentration of lead(ppm).

Group	No	Pb Mean ±SD
Patient	70	0.48±0.14 a
Control	50	0.35±0.09 b
LSD		0.05

Figure 2: The lead levels in patient and control groups.

Zinc 33.1:

The results showed that a significant decrease in the concentration of zinc in PCO patients and control groups (p≤0.05). The resultsagreedwith literature.¹⁴ In other studies, the serum zinc level in PCO patients, was low due to association with TG / HDL, which is associated with cardiovascular disease.¹⁵

Table 3 The concentration of zinc (ppm)in patient and control groups.

Group	No	Zn Mean ±SD
Patient	70	0.91±0.20 b
Control	50	1.85± 0.28 a
LSD		0.07

Figure 3: The zinc levels in patient and control.

3.1.4 Iron:

The resultsshowed a significant increase in the concentration of serum ferritinin PCOS in comparison with control groups (p≤0.05). The results agreed with literature.^9,16 The menstrual dysfunction acts as a predictor of high serum ferritin in PCOS.¹⁷

Table 4 The concentration of iron (ppm).

Group	No	Zn Mean ±SD
Patient	70	0.91±0.20 b
Control	50	1.85± 0.28 a
LSD		0.07

Figure 4: The iron levels in patient and control groups.

3.2 The levels of trace elements with ageat constant BMI:

The patient groups are divided into three groups: ≤20 years, ≤30 years, and ≥40 years.constant BMI ≥75. Note: Each value represents mean±SD values with non-identical superscript (a and b) for vertical comparison and (A,B) for horizontal comparison are considered significantly differences (p≤0.05).

3.2.1 Nickel:

The results showed thatthe concentration of serum nickelin PCOS patient groups are higher than in controlgroups(p≤0.05).The serum nickel concentration in A, B, and C groups are higher than in AC, BC, and CC groups (p≤0.05) (see Table 5 and Figure 5). In addition, the serum Ni concentration in B and C groups are higher than inA, while BC and CC groups are higher than in AC.

Table 5: The concentration of Nickel.

Patient	Age	Ni Mean ±SD	Control	Age	NiMean ±SD	LSD
A	≤20	0.42± 0.09 ^A,c	AC	≤20	0.30± 0.06^B,b	0.02
B	≤30	0.70± 0.10 ^A,b	BC	≤30	0.38± 0.07^B,a	0.03
C	≥40	0.91±0.05 ^A,a	CC	≥40	0.39 ±0.09 ^B,a	0.05
	LSD	0.09		LSD	0.06

Table 7: The concentration of serum Fe(ppm) according to age.

Patient	Age	Fe Mean ±SD	Control	Age	Fe Mean ±SD	LSD
A	≤20	2.08 ± 0.28 ^{A , a}	AC	≤20	1.36 ±0.20 ^{B ,b}	0.13
B	≤30	2.26 ± 0.56 ^{A ,a}	BC	≤30	1.42 ± 0. 22^B,b	0.16
C	≥40	2.15 ±0.30 ^{A ,a}	CC	≥40	1.59 ±0.19 ^B,a	0.25
	LSD	0.15		LSD	0.10

Figure 7 The concentration of Fe in patient and control groups according to age.

Table 8 The concentration of serum Zn(ppm) according to age.

Patient.	Age	Zn Mean ±SD	Control	Age	ZnMean ±SD	LSD
A	≤20	1.01± 0.25 ^{B ,a}	AC	≤20	1.73±0.10 ^{A, b}	0.12
B	≤30	0.90 ± 0.24 ^{B, b}	BC	≤30	1.96±0.41 ^A,^a	0.10
C	≥40	0.84 ±0.11 ^B,b	CC	≥40	1.68±0.34 ^{A, b}	0.12
LSD		0.08	LSD		0.11

Figures 8: The concentration of Zn in patient and control according to age.

Table 9 The concentration of serum Ni (ppm) according to BMI (Kg/m²).

Patient	Body mass index (BMI)	Ni Mean ±SD	Control	Body mass index (BMI)	NiMean ±SD	LSD
A	Non-obese	0.66±0.14^A,a	AC	Non-Obese	0.31 ±0.01^{B, a}	0.10
B	Obesity	0.72±0.15^A,a	BC	Obesity	0.35±0.01^B,^a	0.13
C	Overweight	0.71±0.31^A,a	CC	Over weight	0.36 ±0.01^{B, a}	0.18
	LSD	0.09		LSD	0.08

Table 10 The concentration of Pb of serum (ppm) according to BMI (Kg/m²)

Patient	Body mass index (BMI)	PbMean ±SD	Control	Body mass index (BMI)	PbMean ±SD	LSD
A	Non-obese	0.46± 0.11 ^{A ,a}	AC	Non-obese	0.30 ±0.06^{B, a}	0.11
B	Obeity	0.47±0.12 ^{A, a}	BC	Obesity	0.40±0.09 ^B,^a	0.05
C	Over wight	0.50± 0.09 ^{A, a}	CC	Over wight	0.34 ± 0.04^B,a	0.07
	LSD	0.08		LSD	0.09

Table 11 The concentration of Fe serum (ppm) according to BMI (Kg/m²).

Patient	Body mass index(BMI)	Fe Mean ±SD	Control	Body mass index(BMI)	Fe Mean ±SD	LSD
A	Non-obese	2.34±0.40 ^A,a	AC	Non-obese	1.55±0.31^{B, a}	0.11
B	Obesity	2.11±0.42 ^{A, a}	BC	Obesity	1.45± 0.17^B,^a	0.17
C	Overweight	2.21±0.51^{A, a}	CC	Overweight	1.52± 0.21 ^B,a	0.16
	LSD	0.14		LSD	0.10

Table 12: The concentration of serum Zn(ppm) according to BMI (Kg/m²).

Pat	Body mass index(BMI)	Zn Mean ±SD	Control	Body mass index(BMI)	Zn Mean ±SD	LSD
A	Non-obese	0.93±0.26 ^B,a	AC	Non-obese	1.67±0.30^{A, b}	0.22
B	Obesity	0.83±0.18 ^B,b	BC	Obesity	1.91±0.49^A,^a	0.11
C	Over wight	0.96±0.24 ^B,a	CC	Over wight	1.99±0.50 ^A,a	0.11
	LSD	0.09		LSD	0.17

4. REFERENCE:

1. Skrgatic L., Baldani DP, Cerne J., Ferk P., and Gersak K.‘CAG repeat polymorphism in the androgen receptor gene is not directly associated with polycystic ovary syndrome but influences serum testosterone levels’.J Steroid Biochem Mol Biol, 2012, 128(3), 107–112

2. Matalliotakis I., Kourtis A., Koukoura O., and Panidis D.‘Polycystic ovary syndrome: etiology and pathogenesis’. Arch Gynecol Obstet.2006, 274(4),187–197.

3 Franks S.‘Polycystic ovary syndrome’. N Engl J Med.1995,333, 853-861

4. Coskun A., Arikan T., Kilinc M., Arikan DC., and Ekerbiçer HÇ. ‘Plasma selenium levels in Turkish women with polycystic ovary syndrome’.Eur J Obstet Gynecol Reprod Biol.2013,168(2), 183–186.

5. Lin M-W, Tsai S-J, Chou P-Y, Huang M-F, Sun HS, and Wu M-H.‘Vitamin D receptor 1a promotor −1521 G/C and −1012 A/G polymorphisms in polycystic ovary syndrome’. Taiwan J Obstet Gynecol.2012, 51(4),565–571.

6. Farrell K., and Antoni MH. ‘Insulin resistance, obesity,inflammation, and depression in polycystic ovary syndrome: biobehavioral mechanisms and interventions’. Fertil Steril. 2010, 9, 1565-1574.

7. Huda M. Ali, and Mohammed A. Al-Zubaidi. "Evaluation of Trace Elements in Iraqi Patients with Rheumatoid Arthritis by using Atomic Absorption Spectrophotometer (AAS)".Iraqi Journal of Pharmaceutical Sciences.2017, 21(2), 77-84.

8. Kurdoglu Z., Kurdoglu M., Demir H, and Sahin H."Serum trace elements and heavy metals in polycystic ovary syndrome".Human & experimental toxicology. 2012, 31(5), 452-456.

9. Guanchao Zheng, Lijun Wang, Zhizhun Guo, LingbinSun, Lingling Wang, Chonggang Wang, Zhenghong Zuo, and Huiling Qiu."Association of serum heavy metals and trace element concentrations with reproductive hormone levels and polycystic ovary syndrome in a Chinese population".Biological trace element research. 2015, 167(1), 1-10.

10. Hashemi F. Abolhasan, and Hakimi M. SM.‘Serum and urine level of zinc in patients with minor beta’Iranian Journal of Pathology. 2007, 2, 54–58.

11. Papageorgiou T., Zacharoulis D., Xenos D.,and Androulakis G.‘Determination of trace elements (Cu, Zn, Mn, Pb) and magnesium by atomic absorption in patients receiving total parenteral nutrition. Nutrition. 2002, 18, 32–34.

12. Sameerah A., Zearah1 Saad S.Hamadie, and Nadhum A.N.Awad1.International Research Journal of Natural Sciences. 2016, 4(1), 1-8.

13. Gallagher CM., Moonga BS., and Kovach JS.‘Cadmium, follicle stimulating hormone, and effects on bone in women age 42–60 years NHANES III’. Environ Res. 2010, 110(1), 105.

14. Guler I., Himmetoglu O., Turp A., Erdem A., Erdem M., Onan MA., Taskiran C., Taslipinar MY.,and Guner H.,‘Zinc and homocysteine levels in polycystic ovarian syndrome patients with insulin resistance’Biol Trace Elem Res. 2014,158(3), 297–304.

15. Balen A., and Rajkowha M.,‘Polycystic ovary syndrome a systemic disorder’ Best Pract Res Clin Obstet Gynaecol. 2003, 17, 263–274.

16. Escobar-Morreale HF., Luque-Ramirez M., Alvarez-Blasco F.,Botella-Carretero JI., Sancho J., andSan Millan JL.,‘Body iron storesare increased in overweight and obese women with polycystic ovary syndrome’. Diabetes Care. 2005, 28, 2042–2044.

17. Martínez-García MA., Luque-Ramírez M., SanMillán JL.,and Escobar-Morreale HF. ‘Body iron stores and glucose intolerance in premenopausal women: role of hyperandrogenism, insulin resistance, and genomic variants related to inflammation, oxidative stress, and iron metabolism’. Diabetes Care. 2009, 32(8),1525-1530.

18. Chen YW., Yang, CY., Huang CF., Hung DZ., and Leung YM., Liu SH. ‘Heavy metals, islet function and diabetes development’. Islets. 2009, 1,169–176.

Received on 08.09.2018 Modified on 24.10.2018

Research J. Pharm. and Tech 2019; 12(9):4447-4453.

DOI: 10.5958/0974-360X.2019.00767.4