دراسة مناعية وجزيئية لمرضى البهاق Vitiligo في محافظة كربلاء

Immunological And Molecular Study of Vitiligo Patients in Karbalaa Province

The study aimed to shed light on vitiligo, through testing and immune genetic, knowing the change in the levels of certain immune variables, and certain hormones and certain genes using different genetic ways in people with vitiligo, as well as knowledge of the relationship between those standards and the extent of vulnerability of those genes under study, included the current study, 50 people from vitiligo patients, as well as 50 people (apparently healthy) as the control group for the study was collected from Al-Husseini Teaching Hospital in the city of Karbala (unit skin counseling) during the time period from 5/1/2015 until 15/1/2016. All patients sera were checked to assess the immunological criteria levels which each of the SOD-1, TNF-α, IL-2, Serotonin and Cortisol using (ELISA), as well as Immunofluorescence assay technology, has also been studying the genetic variations of IL-2RA gene by using PCR-RFLP as well as the study of the genetic variations of TNF-α gene by using PCR-ARMS and PCR-SSCP the results showed the following:

The Patient samples divided by age into five categories(10-19) (20-29)  (30-39) (40-49) (50-59) years and based on this division reached the number and percentage of each category 10 (20.0%) compare with the control, and the search results revealed the vitiligo patients on sex has reached of male patients 42 (84%)  and females 8 (16%)  have also been patients were divided into groups smokers numbered 13 (26%)  and a non-smoking (74%) 37, while the total patient ratios depending on the presence of family history 20 (40.0%)  30 (60.0%)  respectively, while the situation is reversed according to the kinship between parents of patients, and the social situation of patients groups were divided into married and non-married rate 33 (66.0%)  and 17 (17.0%)  respectively, the distribution of patients dependent on the site of injury as follows 30 (60.0%)  spread throughout the body, the patient in the extremities 18 (36.0%) the patient abdominal and back (2.0%) 1 and the injured facial 1 (2.0%)  Finally, results showed patients ratios based on the duration of the disease, which is divided on the totals of (1-80) month amounted have a percentage 16 (32.0 %), from (81-160) percentage (22.0%), 11, from (161-240) a month, and its percentage (12.0%) 6, from (241-320) a month, which represented 6, (12.0%)  from (321-400) showed increase of 11 (22.0%).

Immune aspect of the study of immune parameters recorded increase significantly with age is generally at (P ≤0.05) have emerged and down the age group (10-19) years, with averages showed anti-oxidative stress-type SOD-1 up significantly (0.00) when compared with the control group rose averages values ​​for each of TNF-α, IL-2 to reach the levels (0.32 ± 8.47), (15.25 ± 440.86) (pg / ml), respectively, and the control(0.33±4.91),(3.00±156.53) (pg/ml) respectively while decreased levels of serotonin and cortisol with the averages of the healthy and the highest rates of those standards is significant (0.00) among the age group (50-59 years) for the aged (20.36 ± 72.08) ( 0.35 ± 1.47), respectively, compared with the control, rising SOD-rate of 1 in patients in this age group is significant (0.00) for an average (62.15 ± 810.99) also increased the level of TNF-α and IL-2 and serotonin and cortisol increase significantly (0.00) to (0.59 ±19.58), (34.48 ±744.82) (pg / ml), (0.35±1.47) (ng / ml), (μg / dl) (20.36±72.08), respectively, compared with the rate control group, which was to have those same standards (0.87 ± 9.77), (158.19 ±314.06) (pg / ml), (0.07 ± 0.82) (ng / ml), (3.78±17.38) (μg / dl) respectively. There was also an impact on the type of sex on immunological parameters under study here statistical analysis dealt with the study of male patients with compared with the male control group as it appeared that both the SOD-1,TNF-α, serotonin and cortisol has risen significantly (0.00) in patients compared with healthy group except for IL-2, which fell in a significant (0.73) when the male vitiligo patients compared with the control, also conducted a study a comparison between females vitiligo patients and among females control group as the comparison significant increased results showed in all immunological criteria and serotonin and cortisol at (P ≤0.05) for females vitiligo patients when compared to the same standard rates of female control group.Also, it has been studied the negative role of smoking on vitiligo, has divided the patients samples were deviled in to smokers and non-smokers, as the value of SOD-1 and TNF-α risen significantly in smokers patients to reach (71.06±725.06), (pg / ml) (3.09±12.75) compared with the control group (51.58 ± 225.57), (pg / ml) (1.42±7.25), while IL-2 rate show a significant decrease in the levels (70.64 ± 611.58) (pg / ml) compared with the control group  (379.18±199.15 ) (pg / ml) either serotonin and cortisol increased as compared to healthy controls. significant differences between patients who have a family history of the disease , the results showed the increase of immunological criteria levels of hormones .The results showed a decrease in immune parameters and hormone levels in patients who have a kinship between the parents compared to patients who have kinship between parents has been the declining moral at (P ≤0.05), a case study of social status that deal with being  married than non-married couples. The duration of the disease after the split of appropriation categories months as observed variation SOD-1 on different months those levels, either TNF-α highest rate was recorded among the group of (161-240) a month as it was (3.17±17.07) (pg / ml) and finally fell to the category of (1-80) a month to (1.69±9.46) (pg / ml), and the results posted gains in the averages IL-2 infection among categories of (300-241) a month (77.87±700.07) ( pg / ml), while the rest declined aggregates the serotonin has recorded a significant rise in the group ranging between (1-80) a month (67.36±672.58) (ng / ml), while other rates varied with different other groups, as is the case of cortisol .It showed Find a correlation significant results at (P ≤0.05) between immune parameters under study in general, while the results of the study showed correlations between immunological parameters depending on age and duration of the disease as it appeared a correlation significant at (P ≤0.01) and (P ≤0.05) and then gradually start to lose correlation with aging and progress injury. Results have been found amplification of IL -2RA gene RFLP-PCR was found that the molecular weight of the gene IL-2RA is 180pb The results of the genetic variation it has emerged that the pattern B is the most frequency also found that type B is the most frequency compared with type A but the reluctance to less than the control in mental patients. Magnified cut nucleotide gene (308) ARMS-PCR and by using SSCP-PCR technique and after migration to central acrylamide been getting the results it is found that 3AA pattern which represents the three packages is the most frequency in patients with vitiligo while 4BB style that represents the four packages It is the lowest frequency in patients. In light of the demonstrated by the results infer the existence of a direct correlation to the age of the immune standards and levels of hormones as well as having the effect of each sex, smoking and kinship between the boys and the duration of infection is on those standards as well as the presence of hereditary genes, which include all of the IL-2RA and Jane TNF-α for vitiligo patients.