Vitamin D Status and its Receptor Genes BsmI, FokI, ApaI, TaqI Polymorphism in Relation to Glucose Metabolism in Obese Iraqi Type 2 Diabetes Mellitus Patients

Diabetes Mellitus of type-2 is known to be one of main chronic diseases of related to non-communicable. Moreover, is there are any of the complications noticed, it be the pathway to immediate death in a nationwide space. This is one of the main reasons why the world is losing so many people with the complication of the disease. It can be said that, worldwide, more than 285 people in millions are suffering from diabetes. Most of the people are suffering from the type 2 of the diabetes according to the International Diabetes Federation, which can be abbreviated as IDF in the year, 2011. In the present time, the lack of Vitamin D is considered to be an issue that is related to the public health around the globe. By the year 2008, it was accounted that more *Corresponding author: Dr. Hassan F. Al-Azzawie, College of Science, Baghdad University, Iraq, Tel: 964770 7160860; E-mail: hazzawie2@yahoo.com


Introduction
Diabetes Mellitus of type-2 is known to be one of main chronic diseases of related to non-communicable. Moreover, is there are any of the complications noticed, it be the pathway to immediate death in a nationwide space. This is one of the main reasons why the world is losing so many people with the complication of the disease. It can be said that, worldwide, more than 285 people in millions are suffering from diabetes. Most of the people are suffering from the type 2 of the diabetes according to the International Diabetes Federation, which can be abbreviated as IDF in the year, 2011. In the present time, the lack of Vitamin D is considered to be an issue that is related to the public health around the globe. By the year 2008, it was accounted that more Page 2 of 10 with Type 2 of diabetes. This has been clearly visible with the records that have been shown in the reports by the health organization [1,2]. Clinical reviews have demonstrated a constructive relationship between flowing vitamin D (25-hydroxyvitamin D; 25(OH)D) levels and insulin affectability, showing that vitamin D inadequacy may incline individuals to glucose narrow-mindedness, adjusted insulin emission and sort 2 diabetes [3]. This is done either through direct activity by means of vitamin D receptor (VDR) initiation or by implication by means of both calcemic hormones and irritation [4,5]. Chronic inflammation is often observed or seen in the people who are obese. Their body is associated with the building of the Resistance of insulin. This gives rise to the risk of growing a deficient [6] of diabetes of type 2. Hotamisligil is the first one who formed a link within the obesity, insulin along with inflammation [7]. It is quite evident that the activity of provocative pathways interferes with the metabolism which is normal and also causes disruption the proper signaling of the insulin. This increases the expression of the pro-inflammatory cytokines [8]. The cytokines have the target of attacking the receptors cell membranes, responding to the inflammatory resistance along with the exacerbating insulin resistance [9]. Another critical sub-atomic go between that connections pro-inflammatory cytokine to hindrance of insulin flagging is silencers of cytokine flagging (SOCS) 1 and 3, actuated by IL-6, which prompt ubiquitinylation and debasement of insulin receptor substrate (IRS) proteins [10]. The number of evidences that is increasing from time to time; clinical as well as studies from observation with systematic review states that the low-intensity and sub clinical inflammation reaction exists. This even precedes the T2DM of the situation [11][12][13].
The incendiary markers that have demonstrated the most grounded prescient limit in the advancement of T2DM are C-receptive protein (CRP) and IL-6 [11,14]. Epidemiological reviews have demonstrated that TNF-a, CRP, and IL-6 are emphatically related with BMI and muscle to fat quotient [15,16]. A few late human reviews have related vitamin D status with sort 2 DM improvement [13]. In spite of the fact that the mechanisms are essential the part of vitamin D as a hazard ( Figure 8) a DM stays unexplained. It was demonstrated that 25(OH) D3 could be identified with the event of diminished insulin discharge and affectability [17] weight [18] diabetes, and hypertension [19]. Vitamin D receptor (VDR) is circulated in more than 38 tissues, where it apparently controls essential qualities identified with bone metabolism, oxidative harm, ceaseless ailments and irritation [20]. Vitamin D and its receptor complex assume a part in managing B-cell insulin emission and a few polymorphisms, for example, BsmI, FokI, TaqI and ApaI have been portrayed in the VDR qualities that can modify the action of VDR protein [21]. All VDR polymorphisms are situated between exons 8 and nine aside from that FokI is in exon 2. It has been shown that some of these polymorphisms are related to sorting two diabetes mellitus, insulin discharge [22,23] and also to metabolic changes identified with stoutness [21]. Few reviews have inspected VDR quality polymorphisms for association with the danger of these scatters [21,24]. Human reviews researching the impact of vitamin D status with VDR hereditary SNPs on provocative biomarkers of subjects with or at high danger of creating sort two diabetes and its complexities are rare and have produced clashing outcomes. As far as anyone is concerned, no distributed information is accessible concerning the conceivable interaction amongst vitamin D and the insusceptible/fiery go-betweens in sort two diabetes mellitus in Iraq. To date, be that as it may, it stays to be elucidated whether the second-rate inflammation observed in type two diabetes mellitus may be impacted by the insusceptible properties of Vitamin D. Consequently, this review was intended to analyze the part of Vitamin D status in low-power constant aggravation and insulin resistance in T2DM. Also, we proposed to explore the relationship of VDR quality polymorphisms (BsmI as a hazard consider) in DM parts Iraqi patients. It is likewise obscure whether vitamin D status is identified with lipids digestion, and whether the relationship between plasma vitamin D and insulin resistance and also insulin emission is as yet critical in existing T2DM. In this way, the global target of this paper has been revealed insight into these uncertainties in Iraqi individuals. The particular objectives have been: To gauge the pervasiveness of vitamin D inadequacy and the connection between vitamin D status with glycemic control in existing T2DM. In addition DNA genotyped for polymorphism of VDR gene at position sites of FokI, BsmI ApaI and TaqI by PCR-RFLP to confirm its correlation with T2DM patients. Management and screening of the chosen patients was done on the basis of the guidelines of American Diabetes Association. A total of 75 persons with sex and age-matched, obese, normal and healthy as the controls were then enrolled for this study. They were enrolled as chosen from the healthy staff of the stated institute and university with 32 females and 43 males. Their mean ages, in this case ranged between 37 to 61 years and with a mean value ± S.D. of 47.96 ± S.D. of 5.61 years. They were matched on socioeconomic background, BMI, age, sex and no evidence of DM was found. The patients were screened under the standard oral glucose tolerance test. Then questionnaire was formed on lifestyle, socio-demographic information, and geographical origin, past medical history, family history, existence of hypertension, type-2 diabetes and the treatment use. Anthropometric measurements such as body mass index, age, gender, weight, height, and systolic and diastolic blood pressure were undertaken for every volunteer.

Inclusion criteria
These patients were considered as cases with diabetic mellitus type 2 if they had the following criteria: fasting blood sugar more than 140 mg/dl post-prandial sugar more than 200 mg/dl, impaired OGGT and elevated HbA 1C .

Exclusion criteria
Participants were not included if they were found to have liver or kidney disease or extreme cardio-cerebral vascular disease or cancer. Pregnant participants were also excluded. In addition, participants with Diabetes type 1, with abnormal thyroid function, Vitamin D administration or with any osteoporosis. Patients who did not meet these criteria but who gave a history of T2DM were included in the Blood samples collected from all participants after 12-14 h fasting was divided in two tubes either with EDTA for genotyping and HbA1c determination or without the anticoagulant for serum chemical analyses. Glycemic status, lipid profile, Fasting serum glucose (FBG) and insulin concentrations as well as serum lipid profile and were determined as previously described [3]. Insulin resistance was evaluated by quantitative insulin check index. Circulatory 25(OH) D and intact parathyroid hormone were determined using enzyme immunoassay (Germany). DNA was extracted from anticoagulated blood samples using by using wizard genomic DNA purification kit (Promega-USA) according to the manufacturer's protocol. The BsmI, FokI, ApaI and TaqI polymorphic sites for VDR gene were considered. Polymerase chain reaction (PCR) amplification was performed using 3 sets of primersVDR1 (for BsmI), VDR2 (for FokI), and VDR3 (for both ApaI and TaqI) as described according to Garcia D et al. using the thermo cycler. After initial denaturation for 5 min at 95°C, samples were subjected to 30 cycles of amplification, 25 s at 94°C, 20 s at the relevant primer pair annealing temperature and 20s at 72°C. The final step was a 5-min. hold at 72°C. Amplified DNA was digested overnight with suitable amounts of reference restriction enzymes (Fermentas) in the restriction protocol at 37C or 4 h at 55°C according to the manufacturer's instructions. Digestion products were electrophoresed on 2% agarose gel containing 0.4 mg/L ethidium bromide. The polymorphism was documented by photographing under UV illumination.

Polymorphism analysis
All data concerning the 4 restriction enzymes and their restriction patterns are summarized in Table 1. Aliquots of 0.1 U of BsmI, FokI and ApaI and 3U of TaqI restriction enzymes (Fermentas, Lithuania) and 2 μL buffer were added to 4 μL of the VDR PCR products. The alleles were designated as B (650 bp and 175 bp fragments) and b (825 bp fragment) for BsmI, F (196 bp and 69 bp fragments) and f (265 bp) for FokI and A (530 bp and 210 bp) and a (740 bp) for ApaI. TaqI has 2 binding sites on the PCR product, one product a 495 bp and a 245 bp fragment (t) and the other at the single nucleotide polymorphism site in the presence of which the 495 bp fragment will be cut into a smaller 290 bp piece and a 205 bp piece (T).

Statistical analysis
The Hardy Weinberg equilibrium (HWE) equation was utilized to calculate the expected genotype frequency. Allele frequencies of cytokine genes were calculated by direct gene counting methods, while a significant departure from Hardy-Weinberg (H-W) equilibrium was estimated using H-W calculator for two alleles. The Hardy Weinberg equilibrium is the predicted frequencies of genotypes if mating is nonassortative and hardly having mutations from one allele to another. When there are two alleles for a particular gene, A and B, and their respective population frequencies are p and q, then the expected frequencies of the genotypes AA, AB and BB are p², 2pq and q², respectively. Significant differences between the observed and expected frequencies were assessed by Pearson's Chi-square test.
The difference between the expected and observed genotypes was assessed by Chi square X2 test. The P-value<0.05 was measured statistically significant. The frequencies of the alleles and genotypes were evaluated mutually T2DM patient and controlling sets by the Chi square test when appropriate. The odds ratio (OR) was used to estimate the ratio of the risk of Vitamin D status among patient groups with various allele or genotypes compared with the control group. The odds ratio (OR) and 95 percent confidence interval (CI) were also estimated in order to test the relation between T2DM, and investigate means and standard deviations (SD) [25,26].

Results
The clinical characteristics of cases with T2DM and non-diabetic healthy group are summarized in Table 2. No significant differences in mean age and BMI was observed between both groups while patients with T2DM had a significant lower vitamin D level (26.07 ± 13.03 ng/ml vs. 30.28 ± 13.05 ng/ml, p<0.01in healthy control group. Hypo-vitaminosis D was quite prevalent in most Iraqi T2DM patients compared with the healthy non-diabetic group as showed in Table  3. 140 (70.0 percent) of the cases suffered from moderate vitamin D deficiency and 20 (10 percent) had severe deficiency as shown in the Table 2. 15 (7.5 percent) patients were also vitamin D insufficient, leaving only 25 (12.5%) of all the subjects remaining who had sufficient blood vitamin D. There were no significant differences in mean age or mean BMI between DM Type 2 cases and the control (p>0.05). Fasting blood insulin, fasting blood glucose glycosylated hemoglobin, HOMA index, serum triglyceride, serum interleukin-6, interlukin 1-β, tumor  necrotic factor (TNF-α), serum reactive protein and serum adiponectin concentrations all increased significantly in all diabetic subgroups compared with the non-diabetic control, while serum concentration of leptin, apolipoprotein A and osteocalcin decreased significantly in all the diabetic groups compared with the control. Serum-ionized calcium and lipid profile decreased but not significantly in TC, TG, and LDL-C. All serum vitamin D levels were significantly lower in individuals with diabetic type 2 when compared to the controls as seen in Tables 2 and  3. There is no large scaled epid emiology study evaluating vitamin D deficiency in T2DM; however, several studies of different races have reported a high prevalence of hypo-vitaminosis D in patients with T2DM [27,28]. The distribution of vitamin D status in this study sample is very much in line with the study of 29 which found up to 98 percent of the subjects had vitamin D insufficiency or deficiency. Serum vitamin D concentrations in this study are significantly negatively associated with age as expected (r=-0.54 P<0.001). The main resource of vitamin D in our body is sunlight exposure. The elderly usually has decreased outdoor activities, due to the limitation of their self-care ability.
This grim situation and the high prevalence of T2DM diabetes have driven a search for potentially modifiable environmental factors, besides the application of anti-diabetic drugs and modification of life style, in attempting to better manage diabetes. Basic science studies have revealed the plausible effects of vitamin D on the development of diabetes. Coupled with the easy administration and relatively economical price, vitamin D is an ideal candidate for modulating the early development of T2DM. Considering the high prevalence of vitamin D deficiency in established T2DM patients, if vitamin D is still significantly associated with glycemic control, vitamin D as an agent in diabetic management is rather appealing. Data of the present study showed that vitamin D status was significantly associated with HbA1c after adjusting for sex, age, BMI, and duration of diabetes. Data of this work correlates with another study in South Korea which explained the negative relationship between vitamin D status and glycemic control in existing T2DM, which reached a similar conclusion [29]. In addition, evidence from a number of studies has suggested a relationship between vitamin D and micro-vascular complications of T2DM. Moreover, vitamin D has been found to be inversely associated with cardiovascular events and impaired renal function in patients with T2DM [30,31].
The correlation between fasting blood glucose (FBG) and vitamin D status has hardly ever been estimated. Our data also showed a trend of negative correlation with vitamin D. When stratifying patients by BMI, FBG showed a significant inverse relationship with vitamin D, with a standard coefficient beta= -0.066, and p-value<0.01. However, FBG is easily influenced by diet, activities, and anti-diabetic drug, resulting in its frequently fluctuations and nominal value in diabetes management.

Amplification of the VDR gene
Polymerase chain reactions were performed for the amplification of the VDR gene in all diabetic and non-diabetic groups. Agarose gel electrophoresis was used to confirm this amplification for each exon and intron following optimization experiment when sharp, single bands with the accurate molecular sizes were obtained for each exon and intron. The VDR gene was amplified using specific primers for the detection of each FokI‚ BsmI. Taq I and ApaI SNPs. The balance   was usually achieved between reaction components to reduce the nonspecific amplification or to enhance the desired DNA product yield. For this reason, the PCR condition was optimized before starting. In the PCR experiments the optimization of PCR conditions that include annealing temperature and annealing time that was performed in each PCR experiment. In the present study, the optimum annealing temperature for VDR primers was 68ºC for 45 sec. The reaction components were kept at the same concentrations. PCR negative control was performed in each experiment to determine any possible contamination and all negative control samples appeared as empty gel lanes in all experiments. The following figures demonstrate the amplified exons of VDR gene which are: exon 2 (Figure 1), exon 9 ( Figure 2) and intron 8 ( Figures  3 and 4). Amplification was achieved for all patients and controls. Genotypes for these polymorphisms are designated by the first letter of

Page 6 of 10
(FF), heterozygote (Ff) and mutant homozygote (ff) showed one band (265 bp), three bands (265 bp‚169 bp and 96 bp) two bands (169‚96) bp respectively. The genetic polymorphism of VDR was determined at FokI site‚ which were presented with three genotypes (FF‚ Ff‚ ff for FokI in T2DM patients and controls. The genotype FF of VDR gene at FokI position demonstrated a significant (P=0.0001) increased percentage frequency in T2DM patients (70.5 percent) compared to controls (46.6 percent). The F allele was increased (80.25 vs. 56 percent)‚ while f allele was increased in non-diabetic (44 vs. 19.75 percent) in the T2DM patients) and this increase was highly significant (P<0.0001).
The results of PCR-RFLP analysis of the BsmI of the VDR gene in T2DM are shown in Figure 6 and Table 6 which summarized as three genotypes, the wild type homozygote (BB), heterozygote (Bb) and mutant homozygote (bb) showed one band (825 bp), three bands (825 bp‚650 bp and 175 bp) two bands (650 bp and 175 bp) respectively. The genetic polymorphism of VDR was determined at BsmI were presented with three genotypes (BB‚ Bb‚ bb) for BsmI in T2DM patients and controls. For VDR BsmI‚ the BB genotype frequencies were similar in the T2DM patients than the controls (35 percent vs. 32 percent, P>0.05). On the other hand, the Bb genotype frequencies were lower in the T2DM patients compared with the controls (61 percent vs. 66.6 percent, and the difference is not significant (P>0.05). Both B and b alleles frequency have the same value in both T2DM patients and controls, that B allele frequency in T2DM patients is 65.5 percent vs. 65.3 percent in the controls) and the b allele frequency have a similar percentage 34.5 vs. 34.6 percent in T2DM patients and controls respectively, as shown in Table 5.
On the other hand, the results of PCR-RFLP analysis of the TaqI digestion of the VDR gene in T2DM patients and controls are shown in Figure 7 and Table 6 which can be summarized into three genotypes: the wild type homozygote (TT) showed two band (495 bp and 245 bp); heterozygote (Tt) showed four bands (495 bp, 290 bp, 245 bp, 205 bp). Homozygote (tt) showed three bands (290 bp, 245 bp, 205 bp). TaqI has 2 binding sites on the PCR product, one product a 495 bp and a 245 bp fragment (t) and the other at the single nucleotide polymorphism site in the presence of which the fragment 495 bp will cut into a smaller 290 bp piece and a 205 bp piece (T). For VDR digested by TaqI‚ the results show the TT genotype frequencies were significantly higher in the T2DM patients than the controls (39 percent vs. 20 percent, P<0.0034), while the Tt genotype frequencies were lower in the T2DM patients than controls and the difference was not significant (47.5 percent vs.

PCR-RFLP genotyping of VDR gene
The results of PCR-RFLP analysis of the FokI of the VDR gene for T2DM and non-diabetic control subjects are shown in Figure 5 and Tables 4 and 5 which are summarized as the wild type homozygote Page 7 of 10 58.6 percent, P>0.05). Both T and t alleles frequency are significantly different between T2DM patients, compared with controls (P<0.004).
A significant association between the attendance of a VDR FokI F allele and T2DM had been observed‚ while the VDR FokI f allele was more frequent among the control individuals. These finding suggest a protective role for the f allele in contrast to the role of F allele which seems to be a predisposing factor to T2DM in the Iraqi population. In addition, the results seem to reinforce the association of the FF genotype with the susceptibility to the T2DM. On the other hand, the present study did not find any significant differences in VDR gene polymorphism at position BsmI and ApaI between the T2DM patients and the controls‚ an observation that corroborated with a metaanalysis study by Mackawy and Badawi [32] in which the FF genotype association and the BsmI genotypes and allele non-association with T2DM was clearly obtained. Results of this work are inconsistent with Errouagulet [33] who observed that the ff allele was associated with T2DM in Morocco population. However, further studies failed to show an association between this polymorphism and T2DM in a Polish and an Asian meta-analysis study [34][35][36].
Another new finding in this work revealed a significant association between the presence of higher TT genotypes distribution and higher T allele frequency and T2DM‚ while tt genotypes distribution and t allele frequency are more frequent among control individuals. These finding suggest a protective role for the t allele in contrast to the role of T allele which seems to be a predisposing factor to T2DM in Iraqi population for the first time. In addition, the results seem to reinforce the association of the TT genotype with the susceptibility to the T2DM in addition to FF genotype yielded by FokI digestion of VDR gene.
As per the outcomes on allele frequencies and genotype conveyances of the VDR quality TaqI and FokI polymorphisms in patients with T2DM, information in this work concur well with past reviews which announced that allele frequencies and genotype dispersions of the VDR quality FokI polymorphism are essentially unique in T2DM patients, for example, different infections give points of interest. A few polymorphisms, for example, BsmI and FokI, have been depicted in the VDR qualities that can modify the movement of VDR protein as reported by Filus et al. [18]. All VDR polymorphisms are situated between exons 8 and 9 aside from that FokI is in exon 2 [19]. Nonetheless, the hereditary foundation of the illness still stays hazy. Some of these polymorphisms have been appeared to be related with sort 2 diabetes mellitus, insulin emission 20 , and stoutness related

Correlation of VDR (FokI, BsmI, TaqI and Apa I) polymorphisms with metabolic and biochemical, immunological markers
The allele and genotype frequency distribution of VDR (FokI, BsmI, TaqI and Apa I genes among subgroups of T2DM patients according to therapeutic regimes and controls demonstrated significant differences between T2DM patients treated with a combination of oral hypoglycemic drugs and insulin injection in comparison with T2DM patients treated with only oral hypoglycemic drugs and only regarding the genotype and allele distributions of the VDR FokI and TaqI polymorphism. The FF genotype for the VDR (FokI) was significantly more frequent in T2DM patients treated with combination oral hypoglycemic drugs and insulin injection than in T2DM treated oral drugs only and controls (X 2 =5.61, P<0.05 and X 2 =15.13, P<0.0001) respectively. Otherwise results may not find a significant association between T2DM patients treated with combination oral hypoglycemic drugs and insulin injection with T2DM patients treated oral hypoglycemic drugs or lifestyle group compared with non-diabetic controls regarding the VDR (BsmI and ApaI) polymorphism genotype and allele distributions (X 2 =4.92, P>0.05. X 2 =3.13, P>0.05, X 2 =3.38, P>0.05) respectively.
The F allele is more frequent in the T2DM patients treated with combination oral hypoglycemic drugs and insulin injection, than in T2DM patients treated only oral hypoglycemic drug, as contrasted to the power group (p<0.001), odds ratio (OR) and 95 percent CI for the F allele of VDR (FokI)=2.30 (1.37-3.86). On the other hand, results showed a significant difference in T allele frequency between T2DM patients treated with combination oral hypoglycemic drugs and insulin injection than in T2DM patients treated only oral hypoglycemic drug only group as compared to control group (p<0.001 and control groups (P<0.001). Furthermore, we additionally thought about the VDR (FokI, BsmI, TaqI and ApaI) genotypes with various clinical parameters of every single contemplated gathering. Table (Data not shown) present the sharing of all scientific variables according to the VDR genotypes experiential in the control subjects. No alliance between VDR FokI, BsmI, TaqI and ApaI polymorphisms is noticed with all clinical variables, Vitamin D and IL-6 levels. Lipid profile exhibited significant associations with FokI and TaqI only in contrast of BsmI and ApaI VDR polymorphism. Then, the same analyses were performed for all the subgroup; T2DM patients according to the therapeutic regimes observed higher levels of IL-6, TC, TG, and LDL-C with lower HDL-C and vitamin D levels were seen in FF genotype than ff genotype carriers of the VDR (FokI) polymorphism respectively. While BB homozygous for the BsmI VDR variation is not related with lower vitamin D values than found in others. Furthermore, the estimations of FPI, FPG, HbA1c, BMI are comparative among all members, autonomously of nearness or nonattendance of both VDR FokI, BsmI TaqI and Apa I polymorphisms.

Results from
No relationship between VDR BsmI polymorphism was observed with every clinical variable part contemplated, or other biochemical parameters with the exception of vitamin D serum levels which    [21][22][23], little research has been directed to date into VDR quality polymorphisms for relationship with the dangers [37,38].
Then again, the Hardy-Weinberg Equilibrium (HWE) condition examination is performed by ascertaining the allele frequencies and the subsequent expected frequencies of the genotype in view of these. On the off chance that the watched frequencies of genotype are near the normal genotype frequencies figured from the watched allele frequencies, then the populace is in Hardy-Weinberg Equilibrium and allele blends are probably going to be free of each other. While testing for Hardy-Weinberg (H-W) balance uncovered that T2DM patients demonstrated a critical variety in the circulation of VDR FokI genotypes (P<0.0001) this was seen due to differences between the watched and expected frequencies of F/F, F/f and f/f genotypes. Besides, critical contrasts were seen in the control test, in which they and expected genotype frequencies (P<0.001), as show in Table 7.
Page 9 of 10 demonstrated lower huge values in bb transporters. In any case, people conveying the mutant latent homozygous FF genotype of the VDR FokI polymorphism introduced fundamentally higher insulin levels, insulin resistance file (HOMA-IR), IL-6 serum levels, and BMI than people with the homozygous FF genotype (P<0.001). Also, the nearness of FF genotype of VDR FokI polymorphism is related with diminished vitamin D concentrations compared to Ff and ff genotypes (P<0.001).
In a trial to investigate the functional aspects of the four VDR polymorphisms, we measured several clinical and metabolic parameters about the T2DM suffer as well as controls included the purpose. The findings of present study revealed that the VDR gene polymorphisms might power the harshness of diabetic complications chaos. Otherwise, we could not discover momentous relationship stuck between ApaI as well as BsmI VDR and the components of glycemic control in type 2 diabetic Iraqi patients except for vitamin D serum levels. In the present study, lipid profile parameters showed significant association with VDR 2228570 C N T (FokI) polymorphism in T2DM patients treated with a combination of oral and insulin injection, in addition, there were higher TC, TG, LDL-C and lower HDL levels in Ff and FF genotype carriers as well lower vitamin D levels in homozygous recessive VDR 2228570 CNT (FokI) and VDR TaqI) [39][40][41].

Conclusion
In diabetic patients, we noticed significant associations between VDR 2228570 C N T (FokI) polymorphism as well as each of the insulin levels, insulin resistance (HOMA-IR), IL-6 levels, and BMI. Vitamin D levels were lower in homozygous recessive VDR 2228570 CNT (FokI) and VDR 1544410 ANG (BsmI). Moreover, though glycaemia is a major characteristic of T2DM, no affiliation was found between fasting Results showed the AA genotype frequencies were lower in the T2DM patients than the controls (29.5 percent vs. 34.7 percent, P=0.82) while the Aa genotype frequencies were higher in the patients than controls but the difference is not significant (57 percent vs. 53.3 percent respectively, P=0.58). Both A and a alleles' frequency were not significantly different between T2DM patients compared with the controls (P<0.05)‚ and A allele frequency in T2DM patients is 61.3 percent vs. 58 percent in the control group while the a allele frequency is 38.7 in T2DM patients vs. 42% in non-diabetic group as shown in Table 7.
plasma glucose levels and VDR quality polymorphisms assessed in this review. Similarly, Malecki et al. and colleagues have explored BsmI, TaqI, and FokI VDR quality polymorphisms in 548 Polish people with and without T2DM and did not discover any relationship of glycaemia with any of the VDR quality polymorphisms either.
It is evident that hypo vitaminism D has prevailed in Iraqi population with T2DM. Alterations in vitamin D action could be lead to may affect Insulin-sensitivity, β-Cell action or both. Moreover, Data of this study demonstrated association between VDR FokI and Taq-I gene polymorphisms and susceptibility to T2DM in the Iraqi population in contrast to VDR BsmI and Apa I which showed no relationship. The possible action of vitamin D in the pathogenesis of T2DM is far from being completely understood. Additionally, further knowledge on this issue may identify new candidate targets in the treatment and prevention of the disease. Therefore, further investigations on this issue are warranted