Hidayah, Nurul (2019) Efek Ekstrak Ethanol Ciplukan (Physalis Angulata) Terhadap Kadar Tgf-Β1 Pada Kultur Fibroblas Keloid In Vitro. Magister thesis, Universitas Brawijaya.
Abstract
Keloid adalah tumor jinak sebagai respon hiperproliferasi berupa pertumbuhan berlebihan dari jaringan fibrosa akibat ketidakseimbangan proses reparasi yang dapat berakibat ke gangguan psikogenik yang mempengaruhi kualitas hidup. Transforming Growth Factor (TGF-ß) memainkan peran utama dalam patofisiologi pembentukan keloid. Tidak ada modalitas terapi tunggal terbaik untuk keloid. Penelitian eksplorasi agen farmakologis untuk keloid terutama yang berasal dari tumbuhan alami semakin diminati dimana senyawa aktif pada tanaman herbal menunjukkan kemampuan antikeloid yang dapat menjadi terapi alternatif potensial tanpa efek samping berat. Senyawa aktif pada ciplukan (Physalis angulata) menunjukkan aktivitas biologis yang luas yaitu antiproliferasi, antiinflamasi, antiimunomodulator, dan lainnya. Pada keloid terdapat peningkatan selularitas dermal, rasio kolagen tipe I dan II serta ekpresi dermal beberapa matriks ekstraselular. TGF-ß1 sebagai modulator utama fibrosis dimana pensinyalan dihasilkan melalui protein Smad, akan menstimulasi proliferasi fibroblas keloid dan sintesis kolagen, menghambat aktivitas kolagen-degradasi matriks metalloproteinase (MMP) serta menginduksi molekul efektor lainnya. P.angulata menunjukkan kandungan flavonoid, alkaloid, phenol, physalin, withanolide, dan lain-lain. Studi terdahulu menunjukkan pemberian senyawa withangulin dari P.angulata pada sel stellata berpotensi sebagai anti fibrogenik. Oleh karena itu diperlukan penelitian mengenai efek ekstrak ethanol P.angulata dalam mensupresi aktivitas fibroblas keloid melalui jalur sinyal TGF-ß / Smad dengan mengukur kadar TGF-ß1 secara in vitro dimana penelitian ini tidak dapat dilakukan pada hewan coba. Penelitian ini bertujuan untuk mengukur kadar TGF-ß1 pada media kultur fibroblas keloid yang diberikan ekstrak ethanol P.angulata dengan berbagai konsentrasi secara in vitro. Studi eksperimental laboratorik yang dilakukan di Poliklinik Kulit dan kelamin RSUD Dr. Saiful Anwar Malang dan Laboratorium Biomedik Fakultas Kedokteran Universitas Brawijaya Malang dengan sampel berupa jaringan keloid dari 1 orang pasien yang didiagnosis keloid, memenuhi kriteria eksklusi dan inklusi serta menandatangani inform consent. Kemudian dilakukan bedah eksisi shave terhadap jaringan keloid pasien, selanjutnya jaringan dikultur pada media terkondisi dan dilakukan subkultur sebanyak tiga kali. Ekstrak ethanol seluruh bagian P.angulata diperoleh dari Laboratorium Farmakologi FK Unibraw. Selanjutnya pada kultur fibroblas keloid diberikan 3 macam dosis yang ditentukan berdasarkan penelitian terdahulu dan disesuaikan hasil MTT assay yaitu 0,25 μg / mL, 0,5 μg / mL dan 1 μg / mL. Setelah inkubasi 24 jam dari supernatan fibroblas keloid kelompok perlakuan dan kontrol negatif dilakukan pengukuran kadar TGF-ß1 dengan metode ELISA Sandwich. Rerata kadar TGF-β1 dari sampel kelompok fibroblas kontrol 124,34±3,22 ng/ml sedangkan rerata kadar TGF-β1 dari sampel kelompok fibroblas dengan tambahan ekstrak ethanol P.angulata dosis 0,25 μg/ml yaitu 150,63±30.29 ng/ml, rerata dosis 0,5 μg/ml yaitu 129,83±14,59 ng/ml dan rerata dosis 1 μg/ml yaitu 154,94±22,77 ng/ml. Oleh karena distribusi data tidak normal, analisis data menggunakan uji Kruskal Wallis dan kadar TGF-β1 tidak berbeda signifikan pada kelompok perlakuan dan kontrol negatif (p>0,05). Hal ini mungkin dapat disebabkan bahwa ekstrak ethanol P.angulata memberikan efek anti inflamasi dan imunomodulator terhadap fibroblas keloid. Kemudian perbedaan konsentrasi ix pada penelitian fitofarmaka juga dapat menimbulkan efek yang berbeda. Selain itu ekstrak ethanol P.angulata juga menghambat ikatan reseptor TGF-β1 sehingga TGF-β1 tidak dapat masuk ke intrasel. Kemudian pada patogenesis keloid melibatkan banyak jalur pensinyalan. Kesimpulan dari penelitian ini adalah kadar TGF-β1 pada kelompok perlakuan lebih tinggi dibandingkan kontrol. Perlu studi lebih lanjut dengan rentang dosis yang lebih sempit, menggunakan senyawa aktif yang lebih dominan dan spesifik serta berefek anti proliferatif yaitu withangulin dengan metode yang juga dapat menilai kadar TGF-β1 intraselular. Efek P.angulata terhadap fibroblas keloid pada jalur intraselular seperti Smad, PAI-1 dan sintesis kolagen juga dapat dipertimbangkan untuk diteliti lebih lanjut.
English Abstract
Keloid is a benign tumor as a hyperproliferation response in the form of excessive growth of fibrous tissue due to an imbalance of the repair process that can result in psychogenic disorders that affect quality of life. Transforming Growth Factor Beta (TGF-ß) plays a major role in the pathophysiology of the formation of keloid. There is no single therapeutic modality that works best for all types of keloids. There is growing interest in explorative studies on pharmacological agents for keloids, particularly agents from natural plants that contain antioxidant, anti-proliferative, and anti-inflammatory properties. These types of plants can potentially become alternative medication without severe side effects for keloids. The active compounds in ciplukan (Physalis angulata) showed extensive biological activity namely anti-proliferation, antiinflammatory, antiimmunomodulatory, and others. In keloids, there is an increase in dermal cellularity, the ratio of type I and II collagen, and dermal expression of several extracellular matrices. TGF-ß1 as the main modulator of fibrosis where the superfamily member signals are produced through Smad protein, will stimulate proliferation of keloid fibroblasts and collagen synthesis, inhibit collagen-degradation activity of matrix metalloproteinase (MMP) and induce other effector molecules. P.angulata show flavonoids, alkaloids, phenols, physalin, and withanolide content as well as other substances. Previous study shows that administration of withangulin from P.angulata in stellate cells has antifibrogenic potential. Therefore, there is a need for research on effects of P.angulata ethanol extract in suppressing keloid fibroblast activity through the TGF-ß/ Smad signaling pathway by measuring TGF-ß1 levels in vitro which may not be carried out to animals. This study aimed to measure the level of TGF-β1 on keloid fibroblast culture media given various concentrations of P.angulata ethanol extract in vitro. This laboratory experimental study was conducted at the Dermatology and Venerology Polyclinic of Dr. Saiful Anwar Malang Hospital and Laboratory of Biomedicine of the Faculty of Medicine Universitas Brawijaya. The samples were keloid tissue from 1 patient diagnosed with keloid which met the exclusion and inclusion criteria and had signed informed consent. The procedure was to perform shave excision surgery on the keloid tissue of the patient. The tissue was then cultured on conditioned media and subcultures were carried out three times. Ethanol extract from all parts of P.angulata was obtained from the Laboratory of Pharmacology, Faculty of Medicine Universitas Brawijaya. Furthermore, keloid fibroblast cultures were given 3 types of dosage determined based on previous studies and MTT assays namely, 0.25 μg / mL, 0.5 μg / mL and 1 μg / mL. After a 24-hour incubation of the keloid fibroblast supernatant in the treatment group and the negative control, TGF-β1 levels were measured using the ELISA Sandwich method. Average TGF-β1 level from the control fibroblast group was 124.34 ± 3.22 ng/ ml while mean of TGF-β1 level from the fibroblast group sample with 0.25 μg/ ml, 0.5 μg/ ml and 1 μg/ ml of ethanol extract P.angulata was 150.63 ± 30.29 ng/ ml, 129.83 ± 14.59 ng/ ml and 154.94 ± 22.77 ng/ ml consecutively. Because the data were not distributed normally, the result of Kruskal Wallis test, the data analysis method, did not show significant difference between TGF-β1 levels of the treatment groups and negative controls (p> 0.05). This is probably due to the fact that P.angulata ethanol extract provides anti-inflammatory and immunomodulatory effects on keloid fibroblasts. Besides that, the different concentration in the fitopharmaca study can also result in different effects. In addition, the P.angulata ethanol extract also inhibits the TGF-β1 receptor bond so that TGF-β1 cannot enter the intracellular. xi Furthermore, in the pathogenesis, keloids involve many signaling pathways that may cause overlapping between one another. The conclusion is that level of TGF-β1 in treatment groups higher than control groups. Future researchers should conduct further study with a narrow range of doses which involves more dominant and specific active compounds and anti-proliferative effects using methods that can assess intracellular TGF-β1 levels. Future researchers should also conduct further investigation on effect of P.angulata on keloid fibroblasts in the intracellular pathways such as Smad, PAI-1 and collagen synthesis.
Other obstract
-
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/615.323 952/HID/e/2019/041902950 |
| Uncontrolled Keywords: | TGF-β1, fibroblas keloid, ciplukan (Physalis angulata),-TGF-β1, keloid fibroblast, ciplukan (Physalis angulata) |
| Subjects: | 600 Technology (Applied sciences) > 615 Pharmacology and therapeutics > 615.3 Organics drugs > 615.32 Drugs derived from plants and mikroorganisms > 615.323 95 Drugs derived from spesific plant (Scrophulariales) > 615.323 952 Drugs derived from spesific plants (tomatoes) |
| Divisions: | Profesi Kedokteran > Spesialis Ilmu Kesehatan Kulit dan Kelamin, Fakultas Kedokteran |
| Depositing User: | Endang Susworini |
| Date Deposited: | 10 Jun 2022 07:19 |
| Last Modified: | 10 Jun 2022 07:19 |
| URI: | http://repository.ub.ac.id/id/eprint/190942 |
![[thumbnail of Nurul Hidayah.pdf]](http://repository.ub.ac.id/style/images/fileicons/text.png) |
Text
Nurul Hidayah.pdf Download (7MB) |
Actions (login required)
 |
View Item |