Serdar YİĞİT, Arzu GEZER, Çiğdem Eda BALKAN BOZLAK, Seda ÇELİK, Seyit Ali BİNGÖL

Pseudomonas aeruginosa ile enfekte olmuş ratlarda oluşan yanıklar üzerine bitkisel karışımların etkileri

Amaç: Alkanna tinctoria bitkisinden elde edilen bir merhemin Pseudomonas aeruginosa kaynaklı enfeksiyonlar üzerindeki iyileştirici etkisini araştırmaktır. Gereç ve Yöntem: Bu çalışmada ortalama 180 gr ağırlığında 18 adet erkek ergen sıçan (ortalama yaş 6 hafta) kullanıldı. Hayvanlar 3 gruba ayrıldı. Grup 1: Kontrol grubu (sadece yanık oluşturuldu, herhangi bir tedavi yapılmadı), Grup 2 (P. aeruginosa): Yanık oluşturuldu ve P. aeruginosa ile enfekte edildi, Grup 3 (Krem): Yanık oluşturularak P. aeruginosa ile enfekte edilerek bitki karışımı sabah akşam uygulandı. Anestezi altında sıçanların sırtları tıraş edildi ve özel olarak üretilmiş 1*1 cm çapında çelik çubuk 15 saniye kaynar suda bekletildikten sonra 20 saniye sırtlarına uygulandı. Yanan alan daha sonra ATCC Pseudomonas aeruginosa suşu ile enfekte edildi ve örnekler 24 saat sonra toplandı. Bu alanda bakteri üremesini saptamak için, numuneler bir mikrobiyoloji laboratuvarında kan ve EMB (Eosin Metilen Mavisi) besiyerine inoküle edildi. Aşılamadan sonra hayvanlar ayrı steril kafeslere yerleştirildi ve rastgele üç gruba ayrıldı. Üreme görüldükten sonra 2., 7., 14. ve 21. günlerde sıçanlardan doku ve kan örnekleri alındı. Bulgular: Merhem uygulanan grupta epitelyal rejenerasyon daha belirgindi. Özellikle yanık oluşturup pomad uyguladığımız grupta 2. gün damarlanma dikkat çekiciydi. VEGF seviyeleri merhem grubunda diğerlerine göre daha fazla arttı. Çalışmanın 2. gününde, hem 2. hem de 3. grup örneklerinde ortalama bakteri sayısı 105’tir. Çalışmanın sonunda, 2. gruptaki bakteri sayısı ortalaması artarken, 3. gruptaki bakteri sayısı ortalaması azalmıştır. Sonuç: A. tinctorial'dan elde edilen merhemin epitel dokuyu başarılı bir şekilde onardığı ve kanda artan VEGF'yi modifiye ederek yaraların iyileşmesine katkı sağladığı sonucuna varıldı. Bununla birlikte, bu merhem terapötik kullanım için tavsiye edilmeden önce daha fazla araştırmaya ihtiyaç vardır

Anahtar Kelimeler:

Burn, Alkanna tinctoria, VEGF, Rat, Pseudomonas Aeruginosa

Effects of herbal mixture on burned rat skin infected with Pseudomonas aeruginosa

Purpose: The purpose of this study is to investigate the healing impact of an ointment derived from the Alkanna tinctoria plant upon Pseudomonas aeruginosa-induced infections. Material and Methods: In this study, 18 male adolescent rats (mean age 6 weeks) weighed an average of 180 g were used. Animals were divided into 3 groups. Group 1: Control group (consisting of burns, no treatment was done), Group 2 (P. aeruginosa): Burn was created and infected with P. aeruginosa, Group 3 (Cream): P. aeruginosa was used to infect the burns area and the herbal mixture was administered twice a day, once in the morning and once in the afternoon. Under anesthesia, the backs of the rats were shaved, and a specially produced steel bar with a diameter of 1*1 cm was immersed in boiling water for 15 seconds before being applied to their backs for 20 seconds. The burned area was subsequently infected with the ATCC Pseudomonas aeruginosa strain, and samples were collected 24 hours later. To detect bacterial growth in this area, the samples were inoculated on blood and EMB (Eosin Methylene Blue) media in a microbiology laboratory. After inoculation, the animals were placed in separate sterile cages and randomly divided into three groups. Once the growth was observed, the tissue and blood samples were harvested from the rats on the 2nd, 7th, 14th, and 21st days. Results: Epithelial regeneration in this group was more prominent. Vascularization was remarkable on the 2nd day, especially in the group in where we induced a burn and applied the ointment. VEGF levels increased more in the ointment group than in that of others. On the 2nd day of the study, the average bacterial count was 105 in sample of both 2nd and 3rd groups. At the end of the study, while the average of bacterial count was increased in the 2nd group, the average of bacterial count was decreased in the 3rd group. Conclusion: It was concluded that the ointment obtained from A. tinctorial successfully repaired the epithelial tissue and contributed to the healing of wounds by modifying increasing VEGF in the blood. However, further research is needed before this ointment can be highly recommended for therapeutic usage

Keywords:

Alkanna tinctoria, Pseudomonas Aeruginosa, Burn, VEGF, Rat,

PDF

___

Holmes, D. I.; Zachary, I., The vascular endothelial growth factor (VEGF) family: angiogenic factors in health and disease. Genome Biol. 2005;6:209.
Arroyo, A. G.; Iruela-Arispe, M. L., Extracellular matrix, inflammation, and the angiogenic response. Cardiovasc. Res. 2010;86:226-35.
Malik S., Bhushan S., Sharma M., Ahuja P. S., Biotechnological approaches to the production of shikonins: a critical review with recent updates. Crit. Rev. Biotechnol. 2016;36:327-40.
Gümüş K., Özlü Z. K., The effect of a beeswax, olive oil and alkanna tinctoria (l.) tausch mixture on burn injuries: An experimental study with a control group. Complement. Ther. Clin. Pract. 2017;34:66-73.
WHO. WHO traditional medicine strategy: 2014-2023. World Health Organization. 2013.
Elsharkawy E., Elshathely M., Jaleel G. A., Al-Johar H. I. Anti-inflammatory effects of medicinal plants mixture used by Bedouin people in Saudi Arabia. Herba Pol. 2013:59.
Talib W. H., Mahasneh A. M., Antimicrobial, cytotoxicity and phytochemical screening of Jordanian plants used in traditional medicine. Molecules (Basel, Switzerland). 2010;15:1811-24.
Brigham L. A, Michaels P. J, Flores H. E. Cell-specific production and antimicrobial activity of naphthoquinones in roots of lithospermum erythrorhizon. Plant Physiol. 1999;119:417-28.
Papageorgiou V. P, Assimopoulou A. N, Couladouros E. A, Hepworth D, Nicolaou K. C. The chemistry and biology of alkannin, shikonin, and related naphthazarin natural products. Angew Chem. Int. Ed. Engl. 1999;38:270-301.
Papageorgiou V. P, Assimopoulou A. N, Ballis A. C. Alkannins and shikonins: a new class of wound healing agents. Curr. Med. Chem. 2008;15:3248-67.
Desai M. H, Rutan R. L, Herndon D. N. Conservative treatment of scald burns is superior to early excision. J Burn Care Res J. 1991;12:482-4.
Salhi N, El Guourrami O., Rouas L., Moussaid S., Moutawalli A., Benkhouili F. Z et al. Evaluation of the Wound healing potential of cynara humilis extracts in the treatment of skin burns. Evid. Based Complementary Altern. Med. eCAM. 2023;2023:5855948.
Kumar V, Abbas AK, Fausto N. Cellular adaptations, cell injury, and cell death. In: Kumar V, Abbas AK, Fausto N, eds. Robbins and cotran pathologic basis of disease. 7th ed. Philadelphia, PA: Saunders Elsevier. 2005;3-46.
Esfahani H. M., Esfahani Z. N., Dehaghi N. K., Hosseini-Sharifabad A., Tabrizian K., Parsa M et al. Anti-inflammatory and anti-nociceptive effects of the ethanolic extracts of Alkanna frigida and Alkanna orientalis. J. Nat. Med. 2012;66:447-52.
Avsar U., Halici Z., Akpinar E., Yayla M., Avsar U., Harun U et al. The Effects of Argan Oil in Second-degree Burn wound healing in rats. Ostomy Wound Manag. 2016;62:26-34.
Takci H. A. M., Turkmen F. U., Anlas, F. C., Alkan F. U., Bakirhan P., Demir C et al. Antimicrobial activity and cytotoxicity of alkanna tinctoria (l.) tausch root extracts. Karadeniz Fen Bilimleri Dergisi. 2019;9:176-85.
Moustafa A., Atiba A. The effectiveness of a mixture of honey, beeswax and olive oil in treatment of canine deep second-degree burn. Glob. Vet. 2015;14:244-50.
Yazdinezhad A., Monsef-Esfahani, H., Ghahremani M. H. Effect of alkanna frigida extracts on 3t3 fibroblast cell proliferation. Int. J. Pharm. Biol. Sci 2013;3:212-15.
Abdulrhman M., Elbarbary N. S, Ahmed Amin D., Saeid Ebrahim R., Honey and a mixture of honey, beeswax, and olive oil-propolis extract in treatment of chemotherapy-induced oral mucositis: a randomized controlled pilot study. Pediatr. Hematol. Oncol. 2012;29:285-92.
Sengul M., Yildiz H., Gungor N., Cetin, B., Eser Z., Ercisli S. Total phenolic content, antioxidant and antimicrobial activities of some medicinal plants. Pak. J. Pharm. Sci. 2009;22:102-6.
Zhang L., Liu C., Yin L., Huang C., Fan S. Mangiferin relieves CCl4-induced liver fibrosis in mice. Sci. Rep. 2023;13:4172.
Enoch, S., Leaper D. J. Basic science of wound healing. Surgery (Oxford). 2008;26:31-37.
Noorlander M. L., Melis P., Jonker A., Van Noorden C. J. A quantitative method to determine the orientation of collagen fibers in the dermis. J Histochem Cytochem. 2002;50:1469-74.
Shanmugasundaram N., Uma T. S., Ramyaa Lakshmi T. S., Babu M. Efficiency of controlled topical delivery of silver sulfadiazine in infected burn wounds. J. Biomed. Mater. Res. Part A. 2009;89:472-82.
Furie B., Furie B. C. Molecular and cellular biology of blood coagulation. NEJM. 1992;326:800-6.
Nissen N. N., Polverini P. J., Koch A. E., Volin M. V., Gamelli R. L., DiPietro L. A. Vascular endothelial growth factor mediates angiogenic activity during the proliferative phase of wound healing. Am. J. Clin. Pathol. 1998;152:1445-52.
Ma L., Gan C., Huang Y., Wang Y., Luo G., Wu J. Comparative proteomic analysis of extracellular matrix proteins secreted by hypertrophic scar with normal skin fibroblasts. Burns & trauma. 2014;2:76-83.
Inan, A., Şen M., Koca C., Ergin M., Dener C. Effects of Aloe vera on colonic anastomoses of rats. Surg. Pract. 2007;11:60-5.
Yu Y., Chen R., Sun Y., Pan Y., Tang W., Zhang S et al. Manipulation of VEGF-induced angiogenesis by 2-N, 6-O-sulfated chitosan. Acta Biomater. 2018;71:510-21.
Johnson K. E., Wilgus T. A., Vascular endothelial growth factor and angiogenesis in the regulation of cutaneous wound repair. Adv. Wound Care. 2014;3:647-61.