Diabetik sıçan lens ve kan dokusunda oluşan oksidatif stres ve antioksidan enzim düzeylerinin incelenmesi
Investigation on oxidative stress and antioxidant enzymes activitiy in diabetic rat lens and blood tissue
- Tez No: 202015
- Danışmanlar: PROF. DR. NİHAT DİLSİZ
- Tez Türü: Yüksek Lisans
- Konular: Biyoloji, Biyokimya, Moleküler Tıp, Biology, Biochemistry, Molecular Medicine
- Anahtar Kelimeler: Belirtilmemiş.
- Yıl: 2007
- Dil: Türkçe
- Üniversite: Harran Üniversitesi
- Enstitü: Fen Bilimleri Enstitüsü
- Ana Bilim Dalı: Biyoloji Ana Bilim Dalı
- Bilim Dalı: Belirtilmemiş.
- Sayfa Sayısı: Belirtilmemiş.
Özet
Almanya'dan getirilen deney hayvanları çoğaltılarak yedi grup oluşturuldu. Birinci grup kontrol grubu olarak kabul edildi ve deney sonuna kadar bu gruba hiçbir işlem yapılmadı. İkinci gruba 65 mg/kg Streptozotosin uygulanarak (i.p) diabet oluşturuldu. Üçüncü gruba STZ uygulanıp diabet olduğu görüldükten sonra günaşırı 20 mg/sıçan Ginkgo biloba (mabet ağacı) ekstraktı oral gastrik gavaj yardımıyla verildi. Dördüncü gruba STZ uygulanıp diabet olduğu görüldükten sonra günaşırı olarak 21.5 mg/sıçan olacak şekilde Tagetes patula (kadife çiçeği) ekstraktı verildi. Beşinci gruba günaşırı 26.6 mg/sıçan olacak şekilde Gentiana olivieri (afat) ekstraktı verildi. Altıncı gruba, diabet oluştuğu anlaşıldıktan sonra 22.5 mg/sıçan Olea europaea (zeytin) ekstraktı uygulandı. Yedinci gruba STZ uygulandıktan beş gün sonra 20 mg/sıçan Aristolochia maurorum (loğusa otu) ekstraktı uygulandı. Deney süresi boyuca grupların ortalama ağırlık değişimi ve kan glukoz miktarı değişimi ölçülerek kaydedildi. Lens dokusunda malondialdehid (MDA) ve glutatyon GSH) ölçümleri yapıldı. Plazmada MDA, GSH, glukoz, kolesterol, yüksek dansiteli lipoprotein-kolesterol (HDL-C), çok düşük dansiteli lipoprotein-kolesterol (VLDLC), trigliserid (TG), üre, kan üre azotu (BUN), kreatinin (CREA), aspartat aminotransferaz (AST), alanin aminotransferaz (ALT) ve ?-glutamil transferaz (GGT) parametreleri ölçüldü. Hazırlanan eritrosit paketlerinden, eritrosit katalaz (CAT), glutatyon S transferaz (GST) ve glutatyon redüktaz (GR) enzim aktiviteleri ölçüldü. Çıkan sonuçlar istatistiksel olarak Mann-Whitney testine göre yorumlandı. Oluşturduğumuz kontrol grubunda (K) deney sonuna kadar %5.2 oranında bir ağırlık artışı gözlemlenirken, farklı bitki ekstraktları uygulanan ama aynı koşul ve yemlerle beslenen gruplardan Ginkgo biloba grubunda (D/G) % 26.6, Tagetes patula grubunda (D/KA) %11.3, Gentiana olivieri grubunda (D/A) %5.3, Olea europaea grubunda (D/Z) %23.6, Aristolochia maurorum grubunda (D/L) %26.3 ve diabet grubunda (D) %22.3 oranında ağırlık azalması kaydedildi. K grubunun kolesterol değeri 48 mg/dl ölçülürken, D grubu 77 mg/dl, D/G grubu 71 mg/dl, D/KA 63 mg/dl, D/A 63 mg/dl, D/Z 100 mg/dl ve D/L 48 mg/dl olarak ölçüldü. D/L grubu hariç tüm gruplarda bu değerin yükseldiği görüldü. K grubu TG değeri 67 mg/dl olarak ölçülürken, D grubu 170 mg/dl, D/G 179 mg/dl, D/KA 180 mg/dl, D/A 183 mg/dl, D/Z 310 mg/dl ve D/L 131 mg/dl ölçüldü. Tüm gruplarda kontrol grubuna göre anlamlı (p<0.05) bir artışın olduğu görüldü. Kontrol grubu HDL-C değeri 50 mg/dl olarak ölçülürken, D grubu 34 mg/dl, D/G 56 mg/dl, D/KA 47 mg/dl, D/A 37 mg/dl, D/Z 39 mg/dl ve D/L 34 mg/dl olarak ölçüldü. D grubunda K grubuna göre anlamlı bir düşüş gözlenirken, D/G ve D/KA gruplarında D grubuna göre anlamlı bir artış gözlemlendi. Kontrol grubu VLDL-C sonucu 15 mg/dl ölçülürken, D grubu 32 mg/dl, D/G 45 mg/dl, D/KA 36 mg/dl, D/A 43 mg/dl, D/Z 63 mg/dl ve D/L 25 mg/dl ölçüldü. Tüm grupların kontrol grubuna göre anlamlı bir artış gösterdikleri tespit edildi. Kontrol grubu GGT seviyesi 1.6 U/L ölçülürken, D grubu 7.29 U/L, D/G 7.2 U/L, D/KA 8.17 U/L, D/A 6.57 U/L, D/Z 24.17 U/L ve D/L 6.14 U/L ölçüldü. Tüm grupların GGT seviyesi K grubuna göre anlamlı bir artış gösterdi. K grubu AST değeri 120 U/L olarak ölçülürken, D grubu 187 U/L, D/G 113 U/L, D/KA 205 U/L, D/A 469 U/L, D/Z 1183 U/L ve D/L 174 U/L olarak ölçüldü. Tüm grupların K grubuna göre anlamlı bir artış gösterdikleri görüldü. K grubu ALT miktarı 53 U/L olarak ölçülürken, D grubu 132 U/L, D/G 119 U/L, D/KA 147 U/L, D/A 289 U/L, D/Z 386 U/L ve D/L 66 U/L olarak ölçüldü. D/L grubunun ALT seviyesini anlamlı derecede azaltarak K grubuna yaklaştırdığı görülürken, diğer tüm gruplarda bu değerin K grubuna göre yüksek çıktığı görüldü. Kontrol grubunun üre değeri 48 mg/dl ölçülürken, D grubu 100 mg/dl, D/G 85 mg/dl, D/KA 101 mg/dl, D/A 122 mg/dl, D/Z 92 mg/dl ve D/L 76 mg/dl ölçüldü. Tüm gruplarda K grubuna göre anlamlı bir artış olurken, D/L grubu bu artışı azaltarak anlamlı oranda K grubuna yaklaştırmıştır. K grubu BUN seviyesi 22 mg/dl ölçülürken, D grubu 47 mg/dl, D/G 39 mg/dl, D/KA 47 mg/dl, D/A 57 mg/dl, D/Z 47 mg/dl ve D/L 36 mg/dl ölçüldü. Tüm grupların K grubuna göre anlamlı derecede artış gösterdikleri, D/L grubunun ise bu yüksek değeri anlamlı oranda azaltarak K grubuna yaklaştırdığı görüldü. K grubunun CREA değeri 0.51 mg/dl çıkarken, D grubu 0.4 mg/dl,D/G 0.43 mg/dl, D/KA 0.43 mg/dl, D/A 0.48 mg/dl,D/Z 0.42 mg/dl ve D/L 0.34 mg/dl çıktı. D/A grubu hariç tüm grupların CREA değerinin K grubuna göre istatistiksel olarak azaldığı görüldü. Kontrol grubu Lens MDA miktarının 16.75 (nmol/100 mg lens yaş ağırlığı) olduğu, D grubunun 19.1, D/G grubunun 11.5, D/KA grubunun 22, D/A grubunun 22.8, D/Z grubunun 39.55 ve D/L grubunun 21.5 olduğu görüldü. D/G grubunda anlamlı derecede azalmanın olduğu görüldü. Kontrol gurubunun lens GSH miktarı (nmol GSH/100 mg lens yaş ağrılığı) 45.85 iken, D grubu 6.3, D/G grubu 8.78, D/KA grubu 8.9, D/A grubu 9.7, D/Z grubu 7.28 ve D/L grubu 9.75 olarak ölçüldü. Tüm grupların GSH değeri K grubuna göre anlamlı derecede düşük ölçüldü. D/G, D/A ve D/L gruplarının azalan GSH miktarını anlamlı derecede arttırarak kontrol grubuna yaklaştırdıkları görüldü. Kontrol grubu plazma MDA değeri 35.5 nmol iken, D grubu 32.2, D/G grubu 26.7 nmol, D/KA grubu 39.8 nmol, D/A grubu 31.8 nmol, D/Z grubu 41.0 nmol ve D/L grubu 21.0 nmol olarak ölçüldü. Kontrol grubu plazma GSH miktarı 988 nmol ölçülürken, D grubu 1122 nmol, D/G grubu 338 nmol, D/KA grubu 531 nmol, D/A grubu 1450 nmol, D/Z grubu 575 nmol ve D/L grubu 280 nmol olarak ölçüldü. Tip I diabetes mellitusta oksidatif stresin zamanla tüm dokularda artarak tahribat oluşturduğunu ve antioksidan savunmanın bu hasarı kısmen de olsa engellediği sonucuna vardık. Uyguladığımız bitkilerden özellikle Ginkgo biloba (mabet ağacı), Aristolochia maurorum (lohusa otu) ve Gentiana olivieri (afat otu)'nin diabetik komplikasyonları anlamlı derecede azalttığı, Olea europaea (zeytin) ve Tagetes patula (kadife çiçeği)'nın da bazı parametrelerde olumlu sonuçlar verdiğini gördük.
Özet (Çeviri)
Reproducing the test animals that were brought from Germany, seven groups were assigned. The first group was accepted as control group and no treatment was applied to this group until the end of the experiment. The second group was treated with 65 mg/kg streptozotocin (i.p) and diabetes was induced. The third group was treated with STZ and after observing that diabetes was induced, 20 mg/rat Gingko biloba (maidenhair tree) extract was given through gastric gavage every other day. The fourth group was treated with STZ and after observing that diabetes was induced, 21.5 mg/rat Tagetes patula (marigold) extract was given every other day. The fifth group was given26.6 mg/rat Gentiana olivieri (griseb) extract every other day. The sixth group was treated with 22.5 mg/rat Olea europaea (olive) extract, after observing that diabetes was induced. Five days after treating the seventh group with STZ 20 mg/rat Aristolochia maurorum (bithwort) extract was treated. During the course of the experiment, average weight change and blood glucose amount changes of the groups were measured and recorded. Malondialdehyde (MDA) and glutatione (GSH) measurements were carried out in lens tissue. In the plasma, MDA, GSH, glucose, cholesterol, high density lipoprotein-cholesterol (HDL-C), very low density lipoprotein-cholesterol (VLDL-C), triglyceride (TG), urea, blood urea nitrogen (BUN), creatinin (CREA), aspartate aminotransferase (AST), alanin aminotransferase (ALT) and gamma glutamyl transferase (GGT) parameters were measured. Erythrocyte catalase (CAT), glutathione-S-transferase (GST) and glutatione reduktase (GR) enzyme activities were measured from the prepared erythrocyte packs. The results that came out were statistically interpreted according to Mann-Whitney test. In control group (K) that we assigned, while a 5.2% weight increase was observed until the end of the experiment, among the groups which were treated with different plant extracts but which were fed by same conditions and food, 26.6 % weight decrease was recorded in Gingko biloba group (D/G); 11.3% weight decrease was recorded in Tagetes patula group (D/KA); 5.3% weight decrease was recorded in Gentiana olivieri group(D/A); 23.6% weight decrease was recorded in Olea europaea group(D/Z) 26.3% weight decrease was recorded in Aristolochia maurorum group (D/L); and 22.3% weight decrease was recorded in diabetes group (D). While the cholesterol value of group K was measured as 48 mg/dl, the cholesterol value of group D was measured as 77 mg/dl; cholesterol value of group D/G was measured as 71 mg/dl; cholesterol value of group D/KA was measured as 63 mg/dl; the cholesterol value of group D/A was measured as 63 mg/dl, the cholesterol value of group D/Z was measured as 100 mg/dl and the cholesterol value of group D/L was measured as 48 mg/dl. It was observed that excluding D/L group, this value increased in all groups. While TG value of group K was measured as 67 mg/dl, group D was measured as 170 mg/dl, group D/G was measured as 179 mg/dl, group D/KA was measured as 180 mg/dl, group D/A was measured as 183 mg/dl, group D/Z was measured as 310 mg/dl and group D/L was measured as 131 mg/dl. A significant (p<0.05) increase when compared to the control group was observed in all groups. In control group while HDL-C value was measured as 50 mg/dl, it was measured as 34 mg/dl in group D, 56 mg/dl in group D/G, 47 mg/dl in group D/KA, 37 mg/dl in group D/A, 39 mg/dl in group D/Z and 34 mg/dl in group D/L. While a significant decrease was observed in group D when compared to group K, no significant increase was observed in groups D/G and D/KA when compared to group D. While control group VLDL-C result was measured as 15 mg/dl, group D was measured as 32 mg/dl, group D/G was measured as 45 mg/dl, group D/KA was measured as 36 mg/dl, group D/A was measured as 43 mg/dl, group D/Z was measured as 63 mg/dl and group D/L was measured as 25 mg/dl. It was found that all groups indicated a significant increase when compared to control group. While GGT level of control group was measured as 1.6 U/L, group D was measured as 7.29 U/L, group D/G was measured as 7.2 U/L, group D/KA was measured as 8.17 U/L, group D/A was measured as 6.57 U/L, group D/Z was measured as 24.17 U/L and group D/L was measured as 6.14 U/L. GGT level of all groups indicated a significant increase when compared to group K. While AST values of group K was measured as 120 U/L, group D was measured as 187 U/L, group D/G was measured as 113 U/L, group D/KA was measured as 205 U/L, group D/A was measured as 469 U/L, group D/Z was measured as 1183 U/L and group D/L was measured as 174 U/L. It was found that all groups indicated a significant increase when compared to group K. While ALT amount of group K was measured as 53 U/L, group D was measured as 132 U/L, group D/G was measured as 119 U/L, group D/KA was measured as 147 U/L, group D/A was measured as 289 U/L, group D/Z was measured as 386 U/L and group D/L was measured as 66 U/L. While it was observed that ALT level of group D/L significantly decreased and approximated to group K, it was found that in all other groups this value was higher than group K. While urea value was measured as 48 mg/dl in control group, in group D it was measured as 100 mg/dl, in group D/G it was measured as 85 mg/dl, in group D/KA it was measured as 101 mg/dl, in group D/A it was measured as 122 mg/dl, in group D/Z it was measured as 92 mg/dl and in group D/L it was measured as 76 mg/dl. While there was a significant increase in all groups when compared to group K, group D/L reduced this increase and significantly approximated to group K. While BUN level of group K was measured as 22 mg/dl, group D was measured as 47 mg/dl, group D/G was measured as 39 mg/dl, group D/KA was measured as 47 mg/dl, group D/A was measured as 57 mg/dl, group D/Z was measured as 47 mg/dl and group D/L was measured as 36 mg/dl. It was found that all groups indicated a significant increase when compared to group K, and group D/L significantly reduced this high value and it was observed that approximated it to group K. While CREA value of group K came out to be as 0.51 mg/dl, group D came out as 0.4 mg/dl, group D/G came out as 0.43 mg/dl, group D/KA came out as 0.43 mg/dl, group D/A came out as 0.48 mg/dl, group D/Z came out as 0.42 mg/dl and group D/L came out as 0.34 mg/dl. It was observed that CREA value of all groups except D/A group statistically decreased when compared to group K. It was found that Lens MDA amount of control group was 16.75 (nmol/100 mg lens wet weight) group D was 19.1, group D/G was 11.5, group D/KA was 22, group D/A was 22.8, group D/Z was 39.55 and group D/L was 21.5. It was observed that there was a significant decrease in group D/G. While Lens GSH of control group (nmol GSH/100 mg lens wet weight) was 45.85, group D was measured as 6.3, group D/G was measured as 8.78, group D/KA was measured as 8.9, group D/A was measured as 9.7, group D/Z was measured as 7.28 and group D/L was measured as 9.75. The GSH value of all groups was measured as were measured as significantly low when compared to group K. It was found that D/G, D/A and D/L groups increased decreasing GSH amount significantly, and approximated them to control group. While plasma MDA value of control group was 35.5 nmol, group D was measured as 32.2, group D/G was measured as 26.7 nmol, group D/KA was measured as 39.8 nmol, group D/A was measured as 31.8 nmol, group D/Z was measured as 41.0 nmol and group D/L was measured as 21.0 nmol. While plasma GSH amount of control group was measured as 988 nmol, group D was measured as 1122 nmol, group D/G was measured as 338 nmol, group D/KA was measured as 531 nmol, group D/A was measured as 1450 nmol, group D/Z was measured as 575 nmol and group D/Lwas measured as 280 nmol. We concluded that oxidative stress increased in Type I diabetes mellitus in time and caused damage in all tissues and antioxidant defense prevented this damage even in part. Among the plants that we treated, we observed that especially Ginkgo biloba (maidenhair tree), Aristolochia maurorum (birthwort) and Gentiana olivieri (griseb) reduced diabetic complications significantly, and Olea europaea (olive) and Tagetes patula (marigold) also had positive results in some parameters.
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