Effect of Rheum ribes Application on Immunohistochemical Localizations of TLR2 and TLR4 in the Liver and Kidneys of Diabetic Rats

Year 2024, Volume: 21 Issue: 3, 148 - 154, 02.12.2024

Ayhan Akgün , Turgay Deprem

https://doi.org/10.32707/ercivet.1587193

Abstract

Diabetes mellitus (DM) causes various complications. The most important of these complications are those occurring in the liver and kidney, which are responsible for hemostasis. Rheum ribes (R. ribes) is used for treatment purposes in DM as well as various diseases. TLR family members are increased in many cases. However, the in-creased expression of TLR2 and TLR4 in diabetic patients is noteworthy. In our study, we aimed to examine the effects of R. ribes extract in diabetes as well as the immunohistochemical localization of TLR2 and TLR4, which are signifi-cantly increased in diabetic cases. Streptozotocin (STZ) was administered intraperitoneally 50 mg/kg in DM subjects. R. ribes and diabetes + R. ribes groups were given 200 mg/kg R. ribes extract by oral gavage for 21 days. In the exam-ination, TLR2 immunoreactivity in liver tissue was found in hepatocytes, central veins, and sinusoid endothelium; TLR4 immunoreactivity was seen in V. centralis, hepatocytes and V. interlobularis. In the kidney, TLR2 and TLR4 immunore-activity was observed in glomerulus cells, proximal tubules, distal tubules, and medulla. While TLR2 was cytoplasmic immunoreactive, TLR4 was both cytoplasmic and nuclear. Immunoreactivity for both TLR2 and TLR4 was observed intensely in the liver and kidney tissues of the diabetes group. But it was observed that immunoreactivity was signifi-cantly reduced in the liver and kidney tissues of the diabetic group administered R. ribes extract. As a result, it was determined that R. ribes plant has a positive effect on the liver and kidney tissues of diabetic rats. Therefore, we think that the R. ribes plant, which is also used in various diseases, will contribute to the development of new treatment methods in the pharmaceutical industry for DM disease.

Keywords

Diabetes mellitus, immunohistochemistry, kidney, liver, Rheum ribes, TLR2, TLR4

Rheum ribes Uygulamasının Diyabetik Ratların Karaciğer ve Böbreklerinde TLR2 ve TLR4'ün İmmunohistokimyasal Lokalizasyonu Üzerine Etkisi

Abstract

Diyabetes mellitus (DM) çeşitli komplikasyonlara sebep olmaktadır. Bu komplikasyonların en önemlileri hemostaz-dan sorumlu olan karaciğer ve böbrekte meydana gelen komplikasyonlardır. Işgın (Rheum ribes, R. ribes) çeşitli hasta-lıkların yanı sıra DM’de de tedavi amaçlı kullanılmaktadır. TLR aile üyeleri birçok durumda artmaktadır. Ancak diyabetik hastalarda TLR2 ve TLR4'teki ekspresyon artışı dikkat çekmektedir. Çalışmamızda R. ribes ekstraktının diyabetteki etkilerinin yanı sıra diyabetik olgularda belirgin olarak artan TLR2 ve TLR4'ün immünohistokimyasal lokalizasyonunu incelemeyi amaçladık. DM deneklerde Streptozotosin (STZ) 50 mg/kg intraperitoneal uygulanarak oluşturuldu. R. ribes ve diyabet + R. ribes gruplarına 21 gün boyunca oral gavaj ile 200 mg/kg R. ribes ekstraktı verildi. Yapılan incelemede karaciğer dokusunda TLR2 immünreaktivitesi hepatositlerde, V. sentralislerde, sinuzoid endotellerinde; TLR4 immünre-aktivitesi V. sentralis, hepatositler ve V. interlobularislerde görüldü. Böbrekte ise TLR2 ve TLR4 immünreaktivite glome-rulus hücrelerinde, proksimal tubullerde, distal tubullerde ve medullada gözlendi. TLR2 sitoplazmik immünreaktif oldu-ğu halde TLR4 hem sitoplazmik hem de nükleerdi. Hem TLR2 hem de TLR4 için immünreaktivite diyabet grubu karaci-ğer ve böbrek dokularında yoğun olarak gözlenirken, R. ribes ekstraktı uygulanan diyabet grubu karaciğer ve böbrek dokularında immünreaktivitenin önemli ölçüde azaldığı gözlendi. Sonuç olarak diyabetik ratların karaciğer ve böbrek dokularında R. ribes bitkisinin pozitif bir etkiye sahip olduğu belirlenmiştir. Bundan dolayı çeşitli hastalıklarda da kullanı-lan R. ribes bitkisi DM hastalığı için ilaç endüstrisinde yeni tedavi yöntemlerinin geliştirilmesine katkı sağlayacağını düşünmekteyiz.

Keywords

Böbrek, diyabetes mellitus, immunohistokimya, karaciğer, Rheum ribes, TLR2, TLR4

References

• Aboonabi A, Rahmat A, Orthman F. Effect of pomegranate on histopathology of liver and kidney on generated oxidative stress diabetic ınduced rats. J Cytol Histol 2014; 6(1): 1-5.
• Akkuş Y, Şıktar E. Rheum ribes (ışkın otu)’in metanol ekstresinin düzenli aerobik yüzme egzersizi uygula-nan rat dokularında antioksidan ve histopatolojik etkileri, Doktora tezi, Atatürk Üniver Kış Sporları ve Spor Bil Enst, Erzurum 2018.
• Asgharian S, Hoseinkhan, H, Bijad E, Lorigooini Z, Rafieian-Kopaei, M. Protective effect of hydroalco-holic Rheum ribes L. extract in male rat model of lead acetate-induced nephrotoxicity. J Nephropat-hol 2018; 7(2): 83-7.
• Dasu MR, Devaraj S, Park S, Jialal I. Increased toll-like receptor (TLR) activation and TLR ligands in recently diagnosed type 2 diabetic subjects. Diabe-tes Care 2010; 33(4): 861-8.
• Devaraj S, Dasu MR, Rockwood J, Winter W, Griffen SC, Jialal I. Increased toll-like receptor (TLR) 2 and TLR4 expression in monocytes from patients with type 1 diabetes: further evidence of a proinflamma-tory state. J Clin Endocrinol Metab 2008; 93(2): 578-83.
• Devaraj S, Tobias P, Kasinath BS, Ramsamooj R, Afify A, Jialal I. Knockout of toll-like receptor-2 attenuates both the proinflammatory state of diabetes and incipient diabetic nephropathy. Arterioscler Thromb Vasc Biol 2011; 31(8): 1796-804.
• Ehses JA, Meier DT, Wueest S, Rytka J, Boller S, Wielinga PY, Donath MY. Toll-like receptor 2-deficient mice are protected from insulin resistance and beta cell dysfunction induced by a high-fat diet. Diabetologia 2010; 53(8):1795-806.
• Gholamhoseinian A, Moradi MN, Sharifi-far F. Screening the methanol extracts of some Iranian plants for acetylcholinesterase inhibitory activity. Res Pharm Sci 2009; 4(2): 105-12.
• Green TJ, Venn BJ, Skeaff CM, Williams SM. Serum vitamin B-12 concentrations and atrophic gastritis in older New Zealanders. Eur J Clin Nutr 2005; 59(2): 205-10.
• Han LP, Li CJ, Sun B, Xie Y, Guan Y, Ma, ZJ, Chen LM. Protective effects of celastrol on diabetic liver injury via TLR4/MyD88/NF-κB signaling pathway in type 2 diabetic rats. J Diabetes Res 2016; 2641248.
• Hsu SM, Raine L, Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: A comparison between ABC and unla-beled antibody (PAP) procedures. J Histochem Cytochem 1981; 29(4): 577-80.
• Kanitkar M, Bhonde R. Existence of islet regenerating factors within the pancreas. Rev Diabet Stud 2004; 1(4): 185-92.
• Karaali ZE, Candan G, Aktuğlu MB, Velet M, Ergen A. Toll-like receptor 2 (TLR-2) gene polymorphisms in type 2 diabetes mellitus. Cell J 2019; 20(4): 559-63.
• Kim DH, Lee JC, Kim S, Oh SH, Lee MK, Kim KW, Lee MS. Inhibition of autoimmune diabetes by TLR2 tolerance. J Immunol 2011; 187(10): 5211-20.
• Konak A, Aktar O. Medikal antropoloji çerçevesinde Tunceli/Ovacık’ta geleneksel sağaltma yöntemleri. CÜ Sosyal Bilimler Dergisi 2009; 35(2): 156-87.
• Kurt M, Atmaca A, Gürlek A. Diyabetik nefropati. Ha-cettepe Med J 2004; 35: 12-7.
• Kürüm A, Kocamış H, Deprem T, ÇINAR M. Determi-ning gene expression profile of GPX 1 in the liver of diabetic rats treated with capsaicin by real-time PCR. Kafkas Univ Vet Fak Derg 2015; 21(2): 259-63.
• Lin M, Yiu WH, Wu HJ, Chan LY, Leung JC, Au WS, Tang SC. Toll-like receptor 4 promotes tubular inflammation in diabetic nephropathy. J Am Soc Nephrol 2012; 23(1):86-102.
• Meral N. Farklı sıcaklık derecelerinin uşkun bitkisinin antioksidan aktivitesi ve fenolik profili üzerine etkisi. YYÜ Tar Bil Derg 2017; 27(1): 88-94.
• Mudaliar H, Pollock C, Komala MG, Chadban S, Wu H, Panchapakesan U. The role of toll-like receptor proteins (TLR) 2 and 4 in mediating inflammation in proximal tubules. Am J Physiol Renal Physiol 2013; 305(2): 143-54.
• Müştak HK, Esendal ÖM. Toll benzeri reseptörler. Etlik Veteriner Mikrobiyoloji Dergisi 2007; 18: 33-8.
• Okihiro MS, Hinton DE. Partial hepatectomy and bile duct ligation in rainbow trout (Oncorhynchus my-kiss): Histologic, immunohistochemical and enzyme histochemical characterization of hepatic regeneration and biliary hyperplasia. Toxicol Pathol 2000; 28(2): 342-56.
• Pickup JC, Williams G. Textbook of Diabetes. Second edition. Wiley-Blackwell 1997; pp. 151-5.
• Rafaat K, Aboul-Ela M, El-Lakany A. Alloxan-induced diabetic thermal hyperalgesia, prophylaxis and phytotherapeutic effects of Rheum ribes L. in mouse model. Arch Pharm Res 2014; 44(8): 1-10.
• Sawa Y, Takata S, Hatakeyama Y, Ishikawa H, Tsu-ruga E. Expression of toll-like receptor 2 in glomerular endothelial cells and promotion of diabetic nephropathy by Porphyromonas gingivalis lipopoly-saccharide. PloS one 2014; 9(5): e97165.
• Sevimli T, Özçelik N. Stz (streptozotosin) ile diyabet oluşturulmuş ratların böbrek dokusunda silibinin’in HIF-1α (hypoxıa-inducible factor-1 alpha) ve TLR-2 (toll like receptor 2) genlerinin mRNA düzeylerine etkisi, Doktora Tezi, SDÜ Sağ Bil Enst, Isparta 2016.
• Yuan S, Liu X, Zhu X, Qu Z, Gong Z, Li J, Liu F. The role of TLR4 on PGC-1α-mediated oxidative stress in tubular cell in diabetic kidney disease. Oxid Med Cell Longev 2018; 2018: 6296802.
• Zhao W, Chen L, Zhou H, Deng C, Han Q, Chen Y, Li S. Protective effect of carvacrol on liver injury in type 2 diabetic db/db mice. Mol Med Rep 2021; 24(5): 741.