Kosala, Khemasili and Prof.dr.M.Aris Widodo,, MS.,SpFK.,PhD and Prof.Dr.dr.Sanarto Santoso,, DTM&H.,SpMK.,(K and Dr.dr.Setyawati Soeharto,, M.kes. (2019) Kajian Efek Ekstrak Metanol Akar Tambolekar (Coptosapelta Flavescens Korth) Sebagai Bronkodilator Dan Antiinflamasi. Doktor thesis, Universitas Brawijaya.
Abstract
Akar tanaman tambolekar (Coptosapelta flavescens Korth dengan sinonim Coptosapelta Tomentosa) secara etnobotani digunakan oleh masyarakat Kalimantan selain untuk mengobati sesak napas atau asma, juga untuk mengobati sakit gigi maupun rematik. Sehingga akar Tambolekar diduga memiliki efek bronkodilator dan antiinflamasi. Sementara bukti ilmiah sebagai bronkodilator tidak ada, dan data mekanisme aksi antiinflamasi juga tidak jelas. Oleh sebab itu penelitian ini bertujuan mengevalusi aktivitas bronkodilator dan antiinflamasi ekstrak metanol akar Tambolekar (Coptosapelta flavescens Korth). Ada tiga tahap penelitian. Penelitian tahap 1 : Mengkaji aktivitas bronkorelaksasi ekstrak metanol akar Tambolekar (EMAT) secara in vitro melalui reseptor kolinergik, reseptor histamin dan beta2 adrenoseptor. Regulasi kontraktilitas otot polos saluran napas telah diketahui melalui reseptor kolinergik, reseptor histamin dan reseptor beta2 adrenergik. Tujuan penelitian tahap 1 mengevaluasi aksi bronkorelaksasi EMAT dengan mekanisme: a) mengantagonis reseptor kolinergik, b) mengantagonis reseptor histamin dan c) menstimulasi beta2 adrenoseptor, pada cincin bronkus guinea pig. Rancangan penelitian tahap 1 adalah eksperimental menggunakan bronkus terpisah guinea pig yang satu sisi dikaitkan pada alat Transducer isometrik pada isolated organ bath, sisi lainnya dihubungkan dengan recorder di computer. Ada tiga kelompok uji : a) cincin bronkus guinea pig dalam organ bath diinkubasi 10 menit dengan 3 dosis berbeda EMAT, lalu di kontraksi dengan metakolin dosis kumulatif; b) cincin bronkus guinea pig diinkubasi 10 menit dengan 3 dosis berbeda EMAT, lalu di kontraksi dengan histamin dosis kumulatif; dan c) cincin bronkus guinea pig diinkubasi dengan 3 dosis berbeda Propranolol, diprekontraksi dengan metakolin dosis tunggal lalu diberi EMAT dosis kumulatif. Data yang diperoleh dibuat Kurva dosis respon (KDR), sehingga diperoleh Emaks dan pD2. Analisa data menggunakan ANOVA dengan p<0,05 dinyatakan berbeda bermakna. Hasil yang didapatkan pada penelitian tahap 1 adalah a) KDR cincin bronkus yang diinkubasi EMAT 2, 4 dan 6 mg/ml dikontraksi dengan metakolin, bergeser kekanan dengan Emaks lebih kecil dan pD2 (-log EC50) lebih kecil dan berbeda bermakna (P<0,05) dibandingkan kontrol; b) KDR cincin bronkus yang diinkubasi EMAT 2, 3 dan 4 mg/ml dikontraksi dengan histamin, bergeser kekanan dengan Emaks lebih kecil dan pD2 (-log EC50) lebih kecil dan berbeda bermakna (P<0,05) dibandingkan kontrol; c) KDR cincin bronkus yang diinkubasi propranolol 0,5 , 1,0 dan 1,5 μM bergeser kekanan dengan Emaks tidak berbeda bermakna (P>0,05) dan pD2 lebih kecil dan berbeda bermakna (P<0,05) dibandingkan KDR cincin bronkus yang tidak diinkubasi propranolol. Kesimpulan EMAT beraktivitas bronkorelaksasi dengan mekanisme: a) mengantagonis reseptor kolinergik secara nonkompetitif; b) mengantagonis reseptor histamin secara nonkompetitif dan c) men stimulasi reseptor beta2 adrenergik, Penelitian tahap 2: menguji aktivitas antiinflamasi EMAT dengan hambatan volume edema kaki tikus Wistar yang diinduksi carrageenan. xi Reson inflamasi fase awal melibatkan sel mast yang melepaskan mediator histamin, serotonin, dan fase lambat pembentukan prostaglandin yang dikatalisir oleh enzim cycloxygenase (COX). Untuk membuktikan akar tambolekar berefek antiinflamasi khususnya pada inflamasi fase awal, dengan pembentukan edema. Dan sekaligus ingin mengetahui EMAT bersifat mencegah atau mengobati inflamasi. Maka tujuan tahap 2 menguji aktivitas antiinflamasi EMAT setelah induksi carrageenan pada kaki tikus Wistar. Rancangan penelitian tahap 2 adalah eksperimental menggunakan tikus Wistar jantan yang diberi EMAT peroral 1 jam sebelum, saat dan 1 jam setelah induksi carrageenan. Pengukuran volume edema kaki tikus menggunakan alat Plethysmometer, dilaksanakan setiap jam setelah induksi carrageenan sampai jam ke 6 dilanjutkan jam ke 24. Analisa data menggunakan ANOVA dengan p<0,05 dinyatakan berbeda bermakna. Hasil yang diperoleh adalah persen hambatan volume edema pada EMAT dosis 1200 mg/kg BB terjadi pada fase awal dan lambat bila pemberiannya 1 jam sebelum diinduksi carrageenan. EMAT dosis 1200 mg/kg BB pada saat dan 1 jam setelah induksi carrageenan, dan dosis 600 mg/kg BB 1 jam sebelum, saat dan 1 jam setelah induksi carrageenan, persen hambatan volume edema terjadi pada fase lambat. Kesimpulan EMAT dosis 600 mg/kg BB beraktivitas antiinflamasi dengan menghambat enzim cyclooxygenase, sementara EMAT dosis 1200 mg/kg BB menghambat pelepasan mediator inflamasi histamin, serotonin bradikinin dan juga menghambat enzim cyclooxygenase. Penelitian tahap 3 : menguji aktivitas antiinflamasi dengan model men stabilkan membran RBC tikus secara invitro. Infiltrasi leukosit terjadi selama respon inflamasi karena perannya sebagai bagian dari pertahanan tubuh terhadap inflamasi. Sel-sel ini melepaskan isi lisosomnya, yang menyebabkan kerusakan jaringan dan inflamasi lebih lanjut, dan memicu pelepasan fosfolipase A2 (PLA2). Isi lisosom neutrofil juga menginduksi degranulasi sel mast sehingga melepaskan histamin pada inflamasi fase awal. Karena membran red blood cell (RBC) memiliki karakteristik yang mirip dengan membran lisosom, dengan menstabilkan membran RBC yang diinduksi hipotonisitas analog dengan menstabilkan lisosom sehingga menghambat enzym lisis seperti PLA2 dan menghambat degranulasi sel mast sehingga menghambat pelepasan histamin. Tujuan tahap 3 untuk mengkaji aktivitas antiinflamasi EMAT dengan menstabilkan membran RBC tikus Wistar yang diinduksi hiposalin. Rancangan penelitian juga eksperimental dengan menggunakan Red Blood Cell (RBC) tikus Wistar yang diberi berbagai konsentrasi EMAT dan Indometasin sebagai kontrol. Kemudian dibaca absorbansinya pada panjang gelombang 560 nm di spektrofotometer. Hasil yang diperoleh adalah EC50 pada EMAT (1,905 ± 0,119 ) mg/ml lebih kecil dan berbeda bermakna ( p<0,05) dibandingkan EC50 Indometasin (10,288 ± 0,212 ) mg/ml. Kesimpulan aktivitas antiinflamasi dengan menstabilkan membran sel darah merah pada EMAT lebih kuat dibandingkan Indometasin, Dari hasil penelitian tahap 1, 2 dan 3 menunjukkan ekstrak metanol akar tambolekar terbukti beraktivitas bronkodilator dengan mekanisme mengantagonis reseptor kolinergik secara nonkompetitif, mengantagonis reseptor histamin secara nonkompetitif dan menstimulasi reseptor beta2 adrenergik pada cincin bronkus guineapig. Dan beraktivitas antiinflamasi dengan menghambat edema kaki tikus yang diinduksi carrageenan, serta menstabilkan membran sel darah merah tikus. Kata kunci :Ekstrak metanol akar Tambolekar (C.flavescens Korth), bronkus guinea pig, bronkorelaksasi, antiinflamasi, carrageenan, stabilisasi membran RBC
English Abstract
The roots of tambolekar plant (Coptosapelta flavescens Korth, synonym: Coptosapelta Tomentosa) are used ethnobotanically by the people in Borneo in addition to treating shortness of breath or asthma, also for treating toothache and rheumatism. So that the Tambolekar root is thought to have bronchodilator and anti-inflammatory effects. While scientific evidence as a bronchodilator does not exist, and the data on the mechanism of anti-inflammatory action are also unclear. Therefore this study aimed to evaluate the bronchodilator and anti-inflammatory activity of Tambolekar root (Coptosapelta flavescens Korth) methanol extract. There were three stages of research Stage 1 study: Test the bronchorelaxation activity of Tambolekar root methanol extract (EMAT) in vitro through cholinergic receptors, histaminic receptors and beta2 adrenoceptors. Regulations for airway smooth muscle contractility are known through cholinergic receptors, histaminic receptors and beta2 adrenergic receptors. The purpose of stage 1 was to evaluate the action of EMAT bronchial relaxation by the mechanism of: a) cholinergic receptor antagonists, b) histaminic receptor antagonists and c) stimulating beta2 adrenoceptor, on guinea pig bronchial ring. The design of the Stage 1 study was experimental using guinea pig isolated bronchus which on one side was connected to the isometric transducer in the isolated organ bath and another side was connected to the recorder on the computer. There were three test groups: a) guinea pig bronchial ring in the organ bath was incubated for 10 minutes with 3 difference EMAT doses, then contracted with cumulative doses of methacholine; b) guinea pig bronchial ring was incubated for 10 minutes with 3 difference EMAT doses then contracted with cumulative doses of histamine; and c) guinea pig bronchial ring was incubated with 3 difference doses of Propranolol, precontracted with a single dose of methacholine and then given cumulative dose of EMAT. The data obtained were presented in dose-response curves (DRC), Emax and pD2. Data analysis used ANOVA with p <0.05 considered as significantly different. The results obtained on stage 1 were a) DRC of bronchial ring incubated with EMAT 2, 4 and 6 mg/ml and contracted with methacholine shifted to the right with smaller Emax and smaller pD2 (-log EC50) and significantly different (P<0.05) compared with controls; b) DRC of bronchial ring incubated with EMAT 2, 3 and 4 mg/ml and contracted with histamine shifted to the right with smaller Emax and smaller pD2 (-log EC50) and significantly different (P<0.05) compared with controls; c) DRC of bronchial ring incubated with propranolol 0.5, 1.0 and 1.5 μM shifted to the right with Emax was not significantly different (P>0.05) but pD2 was smaller and significantly different (P<0.05) compared with the DRC of bronchial ring that was not incubated with propranolol. Conclusion: EMAT has bronchorelaxation activity as: a) non-competitive cholinergic receptor antagonist; b) non-competitive antagonists for histamine receptors and c) stimulating beta2 adrenergic receptor. Stage 2 study: test EMAT anti-inflammatory activity by inhibiting the volume of edema of carrageenan-induced Wistar rat’s feet xiii Early phase of inflammation respons involves mast cells that release histamine, serotonin mediators while late phase involves the formation of prostaglandins catalyzed by the cycloxygenase (COX) enzyme. To prove if tambolekar root has an anti-inflammatory effect, especially in the early phase of inflammation, with edema formation, and at the same time want to know EMAT prevents or treats inflammation. Then the purpose of stage 2 was to determine EMAT anti-inflammatory activity after induction of carrageenan in Wistar rat's feet. The design of stage 2 was experimental using male Wistar rats which were given EMAT orally 1 hour before, during and 1 hour after carrageenan induction. Measurement of volume of rat foot edema were done using Plethysmometer, carried out every hour after carrageenan induction until the 6th hour and continued to the 24th hour. Analysis of data was done using ANOVA with p<0.05 stated as significantly different. The results obtained were: the percent inhibition of edema volume development on EMAT dose of 1200 mg/kg BW occurred both in the early phase and late phase when it was administered 1 hour before carrageenan induction. On EMAT dose of 1200 mg/kg BW administered during and 1 hour after carrageenan induction, and on EMAT dose of 600 mg/kg BW administered 1 hour before, during and 1 hour after carrageenan induction, the percent edema volume inhibition was found in the late phase. Conclusion: EMAT dose of 600 mg/kg BW had anti- inflammatory activity by inhibiting the cyclooxygenase enzyme, while EMAT dose of 1200 mg/kg BW inhibit the release of inflammatory mediators such as histamine, serotonin, and bradykinin and also inhibit the cyclooxygenase enzyme. Stage 3 study: test anti-inflammatory activity with RBC membranes stabilization model in vitro. Leukocyte infiltration occurs during inflammatory response due to its role as part of the body's defense against inflammation. These cells release the contents of their lysosomes, which cause further tissue damage and inflammation, and trigger the release of phospholipase A2 (PLA2). The contents of neutrophil lysosomes also induce mast cell degranulation such that it releases histamine in the early phase of inflammation. As red blood cell (RBC) membrane has similar characteristics as lysosomal membranes, stabilizing RBC membrane induced with hypotonicity is analogous with stabilizing lysosomes which inhibits lysis enzymes such as PLA2, inhibits mast cell degranulation and therefore, inhibits histamine release.The purpose of stage 3 was to study EMAT anti-inflammatory activity through stabilizing the hyposaline-induced RBC membrane of Wistar rats. The study design was also experimental using Red Blood Cell (RBC) of Wistar rats which were given various concentrations of EMAT and Indomethacin as controls. Then their absorbances were read at 560nm wavelength on the spectrophotometer. The results obtained were EC50 of EMAT (1.905 ± 0.119) mg/ml was smaller and significantly different (p<0,05) compared to EC50 of Indomethacin (10.288 ± 0.212) mg/ml. Conclusion: Anti-inflammatory activity of EMAT by stabilizing red blood cell membrane was stronger than that of Indomethacin. Conclusions from the results of stages 1, 2 and 3 showed that tambolekar root methanol extract had bronchodilator activity by noncompetitive cholinergic receptor antagonist mechanism, non-competitive histaminic receptor antagonist and stimulating beta2 adrenergic receptors on the guineapig bronchial ring, and had anti-inflammatory activities by inhibiting carrageenan-induced rat foot edema, and stabilizing rat red blood cell membranes. xiv Keywords: Tambolekar root (C.flavescens Korth) methanol extract, guinea pig bronchus, bronchorelaxation, anti-inflammatory, carrageenan, RBC membrane stabilization.
Other obstract
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| Item Type: | Thesis (Doktor) |
|---|---|
| Identification Number: | DIS/615.323 93/FK/k/2019/061904320 |
| Subjects: | 600 Technology (Applied sciences) > 615 Pharmacology and therapeutics > 615.3 Organics drugs > 615.32 Drugs derived from plants and mikroorganisms > 615.323 93 Drugs derived from specific plants (Gentianales, Plumeria, Rubiaceae) |
| Divisions: | S2/S3 > Doktor Ilmu Kedokteran, Fakultas Kedokteran |
| Depositing User: | Nur Cholis |
| Date Deposited: | 19 Aug 2022 02:23 |
| Last Modified: | 19 Aug 2022 02:23 |
| URI: | http://repository.ub.ac.id/id/eprint/193352 |
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