Publication: Behçet hastalığında erkek cinsiyet hormonu testosteron’un nötrofil aktivitesi üzerindeki etkisi
Abstract
Behçet hastalığı nötrofil hiperaktivitesi ile karakterize, erkeklerde şiddetli seyir gösteren, inflamatuar bir hastalıktır. Çalışmamızda testosteronun hastalık patogenezindeki rolünü göstermek amacıyla, testosteron uyarısı yapılmadan ve uyarıdan sonra nötrofil fonksiyonları in-vitro gözlenmiştir. Behçet hastalarında nötrofil fonksiyonlarına bakabilmek ve bu bulguları kontrol gruplarıyla karşılaştırabilmek için 49 (26E:23K) Behçet, 8 Hirsutzim, 17 (3E:14K) Romatoid Artirit, 16 (8E:8K) Ankilozan Spondilit hastası ve 31 (19E:12K) sağlıklı kontrol grubu ile çalışılmıştır. Oksidatif patlama tayininde DCFH-DA ve PMA kullanılmıştır. Apoptoz protokolü, Phi philux ve propidyum iyodit ile gerçekleştirilmiştir. Aktivasyon belirteci olarak CD66b ve erken apoptoz belirteci olarak CD16 monoklonal antikorları ile flow sitometrik analizler yapılmıştır. Hirsutik kadınların sağlıklı kadınlara oranla yüksek O.P.K (Oksidatif patlama katsayısı) gösterdiği tespit edilmiştir (7.13±4.3 vs 4.29±1.7, p< 0.03). Hasta kontrol gruplarının (RA ve AS) O.P.K'ları hem Behçet hastalarına hem de sağlıklı kontrollere oranla yüksek bulunmuştur (uyaransız: 8.6±4.7 vs. 6.6±4.4, p<0.05; uyaranlı: 9.9±6.5 vs. 5.9±4.3, p<0.01). Testosteron uyaransız, erkek Behçet hastalarının O.P.K'ları kadın Behçet hastalarına oranla yüksek tespit edilmiştir (8.0±4.9 vs. 4.9±3.3, p=0.01). Ancak uyarı sonrasında testosteronun kadın hastalarda nötrofil aktivasyonunu artırma eğiliminde olduğu görülmüştür. Testosteron uyarısı yapılmadan, erkek Behçet hastalarının CD66b ekspresyonları, sağlıklı erkek kontrollere oranla yüksek çıkmıştır (99.1±0.9 vs. 94.7±5.3, p<0.04). Behçet hastalarında apoptoz oranı sağlıklı kontrollere oranla düşük bulunmuştur (5.4±4.0 vs. 12.7±11.0, p<0.008). Bu düşüklüğün kadın Behçet hastalarından kaynaklandığı görülmüştür (uyaransız: 6.3±5.0 vs. 16.5±10.0, p<0.006; uyaranlı: 7.8±5.0 vs. 14.9±12, p<0.04). CD16 yüzey molekülü, testosteron uyarısı sonrası kadın Behçet hastalarında, erkek hastalara oranla yüksek tespit edilmiştir (99±1.3 vs. 98.2±1.7, p<0.04). Bu bulgular testosteronun nötrofil aktivasyonuna etkisi olabildiğini düşündürmektedir. Behçet hastalığında hormonların etkisinin değerlendirilmesi tedavi anlamında da yeni açılımlar sağlayacaktır. EFFECTS OF TESTOSTERONE ON NEUTROPHIL ACTIVITY IN BEHCET'S DISEASE
Behçet's disease (BD) is an inflammatory disease, characterized by neutrophil hyperactivity, also known to be much severe in male patients. To investigate the role of testosterone, in the disease pathogenesis, neutrophil functions were examined both untreated and treated with testosterone, in-vitro. For observing the neutrophil functions and compare them with control groups, 49 patients (23F:26 M) with Behçet's disease, 8 patients with hirsutism , 17 patients with Rheumatoid Arthritis (RA) (14F:3M),16 patients with Ankylosing spondylitis (AS) (8F:8M) and 31 healthy individuals (12F:19 M) were enrolled in the study. DCFH-DA and PMA were used for demonstrating the O.B.I (Oxidtaive burst index) and the apoptosis protoccol had done with Phi philux and propidium iodide. Surface expressions of CD66b and CD16 were analyzed with flow cytometry. Hirsutic patients, had significantly high O.B.I compared to healthy women (7.13±4.3 vs 4.29±1.7, p< 0.03). O.B.I of diseased controls (RA and AS) showed an increase, compared to BD patients and healthy controls (untreated: 8.6±4.7 vs. 6.6±4.4, p<0.05; treated: 9.9±6.5 vs. 5.9±4.3, p<0.01). In BD patients , O.B.I was significantly greater in males without treatment (8.0±4.9 vs. 4.9±3.3, p=0.01). However, following treatment, testosterone was seemed to make a tendency to increase the O.B.I in females. Male patients with BD showed a higher percentage of CD66b expression compared to male healthy subjects without treatment (99.1±0.9 vs. 94.7±5.3, p<0.04). Apoptosis ratio was low in BD patients (5.4±4.0 vs. 12.7±11.0, p<0.008). This difference had mainly originated by female patients (untreated: 6.3±5.0 vs. 16.5±10.0, p<0.006; treated: 7.8±5.0 vs. 14.9±12, p<0.04). The ratio of CD16, early apoptosis marker of neutrophils, was increased in female patients with BD following testosterone treatment compared to male patients (99±1.3 vs. 98.2±1.7, p<0.04). These results indicate that testosterone may effect neutrophil activation. Modulating the effects of hormones can also be a new area for treatment.
Behçet's disease (BD) is an inflammatory disease, characterized by neutrophil hyperactivity, also known to be much severe in male patients. To investigate the role of testosterone, in the disease pathogenesis, neutrophil functions were examined both untreated and treated with testosterone, in-vitro. For observing the neutrophil functions and compare them with control groups, 49 patients (23F:26 M) with Behçet's disease, 8 patients with hirsutism , 17 patients with Rheumatoid Arthritis (RA) (14F:3M),16 patients with Ankylosing spondylitis (AS) (8F:8M) and 31 healthy individuals (12F:19 M) were enrolled in the study. DCFH-DA and PMA were used for demonstrating the O.B.I (Oxidtaive burst index) and the apoptosis protoccol had done with Phi philux and propidium iodide. Surface expressions of CD66b and CD16 were analyzed with flow cytometry. Hirsutic patients, had significantly high O.B.I compared to healthy women (7.13±4.3 vs 4.29±1.7, p< 0.03). O.B.I of diseased controls (RA and AS) showed an increase, compared to BD patients and healthy controls (untreated: 8.6±4.7 vs. 6.6±4.4, p<0.05; treated: 9.9±6.5 vs. 5.9±4.3, p<0.01). In BD patients , O.B.I was significantly greater in males without treatment (8.0±4.9 vs. 4.9±3.3, p=0.01). However, following treatment, testosterone was seemed to make a tendency to increase the O.B.I in females. Male patients with BD showed a higher percentage of CD66b expression compared to male healthy subjects without treatment (99.1±0.9 vs. 94.7±5.3, p<0.04). Apoptosis ratio was low in BD patients (5.4±4.0 vs. 12.7±11.0, p<0.008). This difference had mainly originated by female patients (untreated: 6.3±5.0 vs. 16.5±10.0, p<0.006; treated: 7.8±5.0 vs. 14.9±12, p<0.04). The ratio of CD16, early apoptosis marker of neutrophils, was increased in female patients with BD following testosterone treatment compared to male patients (99±1.3 vs. 98.2±1.7, p<0.04). These results indicate that testosterone may effect neutrophil activation. Modulating the effects of hormones can also be a new area for treatment.