[TP-016]

Trimetazidinin Streptozotosin ile Oluşturulmuş Diyabetik Rat Modelinde Spermetogenesis Üzerine Koruyucu Etkisi Vardır

¹Ankara Atatürk Eğitim ve Araştırma Hastanesi, Üroloji Kliniği, Ankara
²Hacettepe Üniversitesi, Kastamonu Tıp Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, Ankara
³Turgut Özal Üniversitesi, Tıp Fakültesi, Biyokimya Anabilim Dalı, Ankara
⁴İnönü Üniversitesi, Tıp Fakültesi, Üroloji Anabilim Dalı, Malatya

AMAÇ: Çalışmamızın amacı streptozotosin (STZ) ile diyabetik hale getirilmiş ratlarda bir antioksidan madde olan Trimetazidinin (TMZ) koruyucu etkisinin araştırılmasıdır.

YÖNTEM-GEREÇLER: 50 erkek Sprague-Dawley rat randomize olarak beş gruba ayrıldı (Tablo 1): Grup 1 (kontrol), Grup 2 (STZ nedenli diyabetik rat), Grup 3 (oral 1 cc / gün izotonik alan diyabetik rat), Grup 4 (10 mg / kg / gün TMZ tedavisi alan diyabetik rat), Grup 5 (20 mg / kg / gün TMZ tedavisi alan diyabetik rat). Sekiz hafta sonra orşiektomi yapıldı. Tüm gruplarda histopatolojik ve elektron mikroskopik inceleme yapıldı.

BULGULAR: Grup 1 ve 5 te testisin yapısal ve ultrastrüktürel yapısal bulgularının normal olduğu bulundu. Grup 2,3 ve 4 te bozulmuş testiküler yapı ve bozulmuş spermatogeneze sahip bulgular elde edildi (Tablo 2-3, Şekil 1-2).

SONUÇLAR: Uygun ve yeterli dozda TMZ tedavisi uygulanması ratlarda diyabetes mellitusun spermatogeneze olan zararlı etkilerine karşı koruyucudur.

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Trimetazidine Has Protective Effects On Spermatogenesis In A Streptozotocin Induced Diabetic Rat Model

Muhammet Fuat Özcan¹, Emine Rumeysa Hekimoglu², Kemal Ener¹, Mehmet Namuslu³, Ramazan Altintas⁴, Huseyin Tugrul Celik³, Ziya Akbulut¹, Serkan Altinova^1
1Ankara Ataturk Education and Research Hospital, Department of Urology, Ankara, Turkey
²Hacettepe University, Kastamonu Faculty of Medicine, Department of Histology and Embryology, Ankara, Turkey
³Turgut Ozal University, Faculty of Medicine, Departments of Biochemistry, Ankara, Turkey
⁴Inonu University, Faculty of Medicine, Department of Urology, Malatya, Turkey

PURPOSE: The aim of this study was to investigate the protective effects of trimetazidine (TMZ), as an antioxidant agent, on streptozotocin (STZ)-induced diabetic rats.

METHODS: A total of 50 male Sprague-Dawley rats were randomly classified into five groups as follows (Table 1): Group 1 (control), Group 2 (STZ-induced diabetic rats), Group 3 (STZ-induced diabetic rats treated orally with 1 cc/day isotonic saline; Group 4 (diabetic rats treated orally with 10 mg/kg/day TMZ; Group 5 (diabetic rats treated orally with 20 mg/kg/day TMZ. After eight weeks, orchiectomy was carried out. Histopathological and electron microscopic examinations were performed in all groups (Table 2-3, Figure 1-2).

RESULTS: In Groups 1 and 5, the structural and ultra-structural findings of the testicular tissue and spermatogenesis were found normal. In Groups 2, 3 and 4, similar results were obtained in terms of the impaired testicular architecture and degeneration of spermatogenesis.

CONCLUSIONS: The administration of an optimal dose of TMZ protects against the harmful effects of diabetes mellitus on spermatogenesis in rats. TMZ therapy can be used to maintain normal spermatogenesis in diabetic rats.

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Şekil 1: Testis Işık mikroskopi bulguları

Figure 1: Light microscopy findings

Şekil 2: Testis elektron mikroskopi bulguları

Figure 1: Electron microscopy findings

a: MC: Myoid cell, F: Fibroblast, Sc: Sertoli cell, g: spermatogonia, PS: Primary Spermatocyte, ESt: Early Spermatids, star: Late Spermatids, red arrow: Basal lamina and L: lumen (x2000); b: ESt: Early Spermatids, star: Late Spermatids, N: Nucleous, L: lumen, m: mitochondria (x4000); c: Seminiferous epithelium (germinal epithelium) (x8000); d: Interstitial area, Leydig’s cell; N: Nucleous, m: mitochondria, arrow: Ser (x15000); e: Seminiferous epithelium (germinal epithelium); F: Fibroblast, Sc: Sertoli cell, g: spermatogonia red arrow: Basal lamina and L: lumen (x6000); f: seminiferous epithelium (germinal epithelium); N: Nucleous, Ld: Lipid droplet (x6000); g: Seminiferous epithelium (germinal epithelium) (x6000); h: Seminiferous epithelium (germinal epithelium) (x3000); i: ESt: Early Spermatids, star: Late Spermatids, N: Nucleous, m: mitochondria, asretisk: developing acrosomal cap, containing acrosomal granule (arrow) (x6000); j: Interstitial area, Leydig’s cell; Ld: Lipid droplet, MQ: Macrophage, F:Fibroblast (x10000); k: Seminiferous epithelium (germinal epithelium); F: Fibroblast, Sc: Sertoli cell, g: spermatogonia, PS: Primary Spermatocyte, ESt: Early Spermatids, red arrow: Basal lamina and black arrow: electron dense body (x4000); l: ESt: Early Spermatids, star: Late Spermatids, N: Nucleous, L: lumen, m: mitochondria (x6000); m: Seminiferous epithelium (germinal epithelium) (x8000); n: Sc: Sertoli cell, g: spermatogonia, ESt: Early Spermatids, red arrow: Basal lamina and Ld: Lipid droplet, m: mitochondria, V: Vacuole (x8000); o: Interstitial area, Leydig’s cell; Ld: Lipid droplet, MQ: Macrophage, F:Fibroblast (x6000).

a: MC: Myoid cell, F: Fibroblast, Sc: Sertoli cell, g: spermatogonia, PS: Primary Spermatocyte, ESt: Early Spermatids, star: Late Spermatids, red arrow: Basal lamina and L: lumen (x2000); b: ESt: Early Spermatids, star: Late Spermatids, N: Nucleous, L: lumen, m: mitochondria (x4000); c: Seminiferous epithelium (germinal epithelium) (x8000); d: Interstitial area, Leydig’s cell; N: Nucleous, m: mitochondria, arrow: Ser (x15000); e: Seminiferous epithelium (germinal epithelium); F: Fibroblast, Sc: Sertoli cell, g: spermatogonia red arrow: Basal lamina and L: lumen (x6000); f: seminiferous epithelium (germinal epithelium); N: Nucleous, Ld: Lipid droplet (x6000); g: Seminiferous epithelium (germinal epithelium) (x6000); h: Seminiferous epithelium (germinal epithelium) (x3000); i: ESt: Early Spermatids, star: Late Spermatids, N: Nucleous, m: mitochondria, asretisk: developing acrosomal cap, containing acrosomal granule (arrow) (x6000); j: Interstitial area, Leydig’s cell; Ld: Lipid droplet, MQ: Macrophage, F:Fibroblast (x10000); k: Seminiferous epithelium (germinal epithelium); F: Fibroblast, Sc: Sertoli cell, g: spermatogonia, PS: Primary Spermatocyte, ESt: Early Spermatids, red arrow: Basal lamina and black arrow: electron dense body (x4000); l: ESt: Early Spermatids, star: Late Spermatids, N: Nucleous, L: lumen, m: mitochondria (x6000); m: Seminiferous epithelium (germinal epithelium) (x8000); n: Sc: Sertoli cell, g: spermatogonia, ESt: Early Spermatids, red arrow: Basal lamina and Ld: Lipid droplet, m: mitochondria, V: Vacuole (x8000); o: Interstitial area, Leydig’s cell; Ld: Lipid droplet, MQ: Macrophage, F:Fibroblast (x6000).

Grupların Özellikleri

	DM	Oral gavaj	Grup Adı
Grup 1	-	-	Kontrol
Grup 2	+	1 cc izotonik salin /gün	Sham
Grup 3	+	-	DM
Grup 4	+	10 mg/kg/gün TMZ	DM+10 mg TMZ
Grup 5	+	20 mg/kg/gün TMZ	DM+20 mg TMZ

DM: Diyabetes mellitus, TMZ: Trimetazidine,

Table 1: Group characteristics

	DM	Oral gavage	Group name
Group 1	-	-	Kontrol
Group 2	+	1 cc isotonic salin /day	Sham
Group 3	+	-	DM
Group 4	+	10 mg/kg/day TMZ	DM+10 mg TMZ
Group 5	+	20 mg/kg/day TMZ	DM+20 mg TMZ

DM: Diabetes Mellitus, TMZ: Trimetazidine

Table 2: Testiküler dokunun ışık mikroskopi bulguları

	Grup 1	Grup 2	Grup 3	Grup 4	Grup 5
SEMİNİFER TÜBÜL	Normal	Germinal epitelyumda azalmış desquamazasyon, düzensiz seminifer tübül	Germinal epitelyumda azalmış desquamazasyon, düzensiz seminifer tübül	Germinal epitelyumda azalmış desquamazasyon (Grup 2 den daha az), düzensiz seminifer tübül	Normal
Vakuolizasyon	-	+	+	+ (Grup 2 den daha az)	-
Epitel Dejenerasyonu	-	+	+	+	-
Spermatozoa	+	-	-	-	+
Spermatid	+	-	-	-	+
GERMİNAL EPİTELYUM
Basal Membran	Intact	Intact	Intact	Intact	Intact
Tunika Propria	Normal	Normal	Normal	Normal	Normal
Spermatogenezis	Normal	Abnormal	Abnormal	Abnormal	Normal
İNTERSTİSYEL ALAN	Normal	Ödem	Ödem	Kısmen normal, kısmen ödemli	Grup 1 den daha büyük
lokasyon	Normal	Düzensiz	Düzensiz	Düzensiz	Normale yakın
SPERMATOGENEZİS	Normal	Spermatik hücrelerde dejenerasyon	Spermatik hücrelerde dejenerasyon	Spermatik hücrelerde dejenerasyon	Normal

DM: Diabetes mellitus, ST: Seminiferous tubule

Table 4: Light microscopy findings of the testicular tissue in all groups

	Group 1	Group 2	Group 3	Group 4	Group 5
SEMINFEROUS TUBULE	Normal	Reduction and desquamation of the germinal epithelium Irregular ST shape	Reduction and desquamation of the germinal epithelium Irregular ST shape	Reduction and desquamation of the germinal epithelium (less than Group 2) Irregular ST shape	Normal
Vacuolization	Absent	+	+	+ (less than Group 2)	Absent
Degeneration of epithelium	Absent	+	+	+	Absent
Spermatozoa	+	Absent	Absent	Absent	+
Spermatid	+	Absent	Absent	Absent	+
GERMINAL EPITHELIUM
Basement membrane	Intact	Intact	Intact	Intact	Intact
Tunica propria	Normal	Normal	Normal	Normal	Normal
Spermatogenesis	Normal	Abnormal	Abnormal	Abnormal	Normal
INTERSTITIAL AREA	Normal	Edema	Edema	Partly Normal, partly edema	Larger than Group 1
Location	Normal	Irregular	Irregular	Irregular	Almost normal
SPERMATOGENESIS	Normal	Degeneration of the spermatogenic cells	Degeneration of the spermatogenic cells	Degeneration of the spermatogenic cells	Normal

DM: Diabetes mellitus, ST: Seminiferous tubule

Tablo 3: Testiküler dokunun elektron mikroskopik bulguları

	Group 1	Group 2	Group 3	Group 4	Group 5
Seminifer Epitel	Normal	Epitelyal kalınlık ↓	Epitelyal kalınlık ↓	Epitelyal kalınlık ↓	Normal
Spermatogonium	Normal	Normal	Normal	Normal	Normal
Primer spermatocytes	Normal	Stoplazmada büyük ve küçük droplet, küçük hücrelerde hücreler arası bağlantı yok.	Large and small lipid droplet in cytoplasm, Stoplazmada büyük ve küçük droplet, küçük hücrelerde hücreler arası bağlantı yok.	Stoplazmada büyük droplet, küçük hücrelerde hücreler arası bağlantı yok.	Normal, stoplazmada küçük lipit droplet
Erken (round) Spermatid	Normal	Nadir	Nadir	Nadir	Normal
Geç (elongated) spermatid	Normal	Yok	Yok	Yok	Normal
Spermatitte akrozomal vezikül	+	-	-	Nadiren +	+
Sertoli Hücresi	Normal	Normal, sitoplazmada vakuol	Normal, sitoplazmada vakuol	Normal, sitoplazmada vakuol	Normal, nadir vakuol
Leydig hücresi	Çok sayıda hücre zarına bağlı lipid droplet ve büyük miktarda vakuoller	nadiren hücre zarına bağlı lipid droplet, SER ve büyük miktarda vakuol.	nadiren hücre zarına bağlı lipid droplet, SER ve büyük miktarda vakuol.	nadiren hücre zarına bağlı lipid droplet, SER ve büyük miktarda vakuol.	Çok sayıda hücre zarına bağlı lipid droplet ve büyük miktarda SER, nadiren vakuol
Interstitial Alan	artmış vaskülarizasyon	artmış vaskülarizasyon	artmış vaskülarizasyon	artmış vaskülarizasyon	artmış vaskülarizasyon
Spermatogenezis	Normal	Anormal	Anormal	Anormal	Normal

SER: Düz endoskopik Retikulum

Table 3: Electron microscopy findings in testicular tissue

	Group 1	Group 2	Group 3	Group 4	Group 5
Seminiferous (germinal) epithelium	Normal ultrastructure	Epithelial thickness ↓	Epithelial thickness ↓	Epithelial thickness ↓	Normal ultrastructure
Spermatogonium	Normal ultrastructure	Normal ultrastructure	Normal ultrastructure	Normal ultrastructure	Normal ultrastructure
Primer spermatocytes	Normal ultrastructure	Large and small lipid droplet in cytoplasm, Some cells have no intercellular junction	Large and small lipid droplet in cytoplasm, Some cells have no intercellular junction	Large lipid droplet in cytoplasm, Some cells have no intercellular junction	Normal ultrastructure, small lipid droplet in cytoplasm
Early (round) Spermatid	Normal ultrastructure	Rarely	Rarely	Rarely	Normal ultrastructure
Late (elongated) spermatid	Normal ultrastructure	Absent	Absent	Absent	Normal ultrastructure
Acrosomal vesicles in spermatid	+	_	_	Rarely +	+
Sertoli cell	Normal ultrastructure	Normal ultrastructure and vacuole in cytoplasm	Normal ultrastructure and vacuole in cytoplasm	Normal ultrastructure and vacuole in cytoplasm	Normal ultrastructure and rarely vacuole.
Leydig cell	Contains numerous membrane-bound lipid droplets and large amounts of SER	Contains rarely membrane-bound lipid droplets, SER and large amounts of vacuoles.	Contains rarely membrane-bound lipid droplets, SER and large amounts of vacuole.	Contains rarely membrane-bound lipid droplets, SER and large amounts of large vacuole.	Contains numerous membrane-bound lipid droplets and large amounts of SER. Rarely vacuole.
Interstitial Area	Increased vascularization	Increased vascularization	Increased vascularization	Increased vascularization	Increased vascularization
Spermatogenesis	Normal	Abnormal	Abnormal	Abnormal	Normal

SER: Smooth endoplasmic reticulum