Disertasi

Aktivitas antifibrosis ekstrak Turbinaria decuren terkarakterisasi melalui penghambatan TGF1, Smad3 dan MMP13 serta pengaruh terhadap aktivitas decorin pada hati tikus yang diinduksi CCl4. = Antifibrotic Activities of Turbinaria decuren extract characterized by inhibiting TGF1/Smad3, MMP13 and the effect on the activity of decorin in Rat liver induced CCl4.

Pendahuluan: Fibrosis hati yang diakibatkan oleh stres oksidatif ROS dan transformasi faktor pertumbuhan beta 1 (TGF-1) menjadi salah satu target dalam pencegahan dan pengobatan fibrosis hati. Rumput laut coklat Turbinaria decuren diketahui mengandung fukoidan yang bersifat ionik memiliki bioaktivitas berdasarkan spesies dan berat molekul namun belum diteliti secara intensif. Penelitian ini dilakukan dengan tujuan membuktikan efek antifibrosis ekstrak T. decuren terkarakterisasi (eTkar) melalui penghambatan aktivitas TGF-1, MMP 13 dan Smad3 serta pengaruhnya terhadap decorin. Metode: Penelitian dilakukan di Labkesda Provinsi DKI Jakarta, Laboratorium Histologi FKUI, Laboratorium Farmakologi FKUI dan Laboratorium Terpadu FKUI. Ekstrak T.decuren terkarakterisasi (eTkar) hasil penelitian tahap I akan diuji efeknya sebagai antifibrosis melalui mekanisme preventif dan kuratif pada. tikus jantan galur Sprague- Dawley (SD) yang diinduksi dengan CCl4 50% dosis 1 ml/100 gram BB dua kali semingg selama 8 minggu. Enam puluh dua .tikus jantan galur SD dibagi 2 kelompok uji yaitu A: uji preventif dan B: uji kuratif, masing-masing dibagi menjadi 7 kelompok perlakuan secara acak. eTkar diberikan pada dosis 27,5, 55, 110 mg/kg BB p.o. Pada uji preventif eTkar diberikan 1 minggu sebelum dan 7 minggu bersamaan pemberian CCl4. Pada pengujian kuratif diberikan CCl4 selama 2 minggu dan 6 minggu secara bersamaan dengan sampel uji. Pemeriksaan yang dilakukan: ALT, AST, ALP, GGT, MDA, GSH, protein total, TGF-1, Smad3, MMP 13, decorin dan pemeriksaan histopatologi jaringan hati. Sebagai pembanding positif digunakan fukoidan® 50 mg/Kg BB. Hasil: Pada uji preventif dosis eTkar yang mampu menekan luas fibrosis hati tertinggi adalah 27,5 mg/kg BB sebesar 69,89% lebih tinggi dari fukoidan® dan berbeda bermakna dengan kontrol (-). Pada dosis ini juga mampu menekan secara bermakna aktivitas TGF1, MMP 13, ekspresi Smad3, decorin dan meningkatkan kadar GSH secara bermakna dibanding kontrol (-), tetapi tidak bermakna untuk MDA. Pada uji kuratif, eTkar dosis 55 mg/Kg BB mampu menekan persentase luas fibrosis hati sebesar 62,05% yang berbeda bermakna dibanding kontrol (-) (p < 0,05) dan kontrol fukoidan® (p < 0,05). eTkar pada dosis ini juga mampu menekan secara bermakna aktivitas TGF1, MMP 13, ekspresi Smad3, decorin, MDA dan meningkatkan kadar GSH dibanding kontrol (-). Efektivitas antifibrosis eTkar pada kedua metode uji tersebut juga berkorelasi positif dengan parameter antioksidan MDA, GSH, cedera hati (ALT, AST), fungsi hati (ALP, GGT dan albumin), dan derajat fibrosis (luas perlemakan hati, aktivitas TGF1, MMP 13 dan ekspresi Smad3 serta decorin). Aktivitas eTkar sebagai preventif lebih baik dibanding kuratif. Kesimpulan: eTkar terbukti mampu menekan perkembangan fibrosis terutama sebagai preventif pada dosis 27,5 mg/Kg .lebih baik dibanding kuratif pada dosis 55 mg/Kg dan mampu menekan aktivitas MDA, TGF1, MMP 13, ekspresi Smad3, ekspresi, decorin dan biokimia darah terkait fungsi dan cedera hati serta meningkatkan GSH sehingga T. decurrens dari perairan Lampung berpotensi untuk dikembangkan sebagai sediaan antifibrosis.
Kata kunci: Fukoidan, Turbinaria decuren, antioksidan, TGF-1, MMP 13, decorin, Fibrosis hati.


Introduction: Liver fibrosis due to oxidative stress ROS and transforming growth factor beta 1 (TGF-1) became one of the targets in the prevention and treatment of liver fibrosis. The brown seaweed T.decurens fucoidan-containing ionic compounds have a potent bioactive potentials, which depends on species and molecular weight. but still unknown mechanism of action in inhibiting liver fibrosis. This study was conducted in order to prove the effect of the extract antifibrosis characterized by constraints on the expression of protein TGF-1, Smad3 and its effect on protein MMP 13 and decorin. Method: The study was conducted in Labkesda DKI Jakarta province, Histology Laboratory, Pharmacology Laboratory and Integrated Laboratory School of Medicine FKUI. Characterized extracts brown seaweed T.decuren (eTkar) of the results of the phase I study was studied to investigate their effect as antifibrosis in animal model of fibrosis through preventive and curative mechanism on male rats Sprague-Dawley (SD) induced by CCl4 by 50% CCl4 – 1 mL/kg BW twice a week for 8 weeks p.o. Sixty two male rats SD divided into 2 groups, namely A test: test preventive and B: test curative. Each divided into 7 treatment groups at random. eTkar was given a three level doses, i.e 27,5, 55, 110 mg/kg BW p.o. At preventive mechanism, eTkar was delivered for 1 week before and 7 weeks of concurrent CCl4 administration. On curative mechanism, eTkar was delivered the last 6 weeks for 8 weeks CCl4 adminstration simulaneously. The test parameters were biomarkers of liver injury (ALT, AST, MDA and GSH liver), liver function (ALP, GGT and albumin), the degree of fibrosis (fibrosis area, TGF1 and MMP 13 activities, Smad and decorin expression, fatty area hispatologically). Fucoidan® was used as positive control at the dose of 50 mg/Kg BW. Results. In the preventive method, the optimum dose of eTkar capable to suppress liver fibrosis was 27,5 mg/kg BW by 69,89 % of CCl4 group which was better than comercial fucoidan and significanly different. At this dose eTkar was able to significanly suppress the activity TGF1, MMP 13, expression Smad3 decorin and increase GSH level significanly different to (-) group (p < 0,05) but not significanly different on MDA. In the curative method, eTkar at the dose of 55 mg/kg was exhibited 62,05% reduce of liver fibrosis compared (-) group. Curative activity at this dose was significanly higher than fucoidan comercial, different to (-) group (p < 0,05) and capable of inhibiting the TGF1 and MMP 13 activity, Smad3, decorin expression and significanly increase GSH level different to (-) group (p < 0,05) but not for MDA (p=0,771). The antifibrosis efficacy of eTkar in both experiments were also positively correlated with the biomarker of antioxidant activity (MDA & GSH) liver function (hati (ALP, GGT dan albumin), liver injuries (ALT, AST) and degree of fibrosis (fatty change, actvity of TGF1 and MMP 13, expression of Smad3 and decorin). The fibrostatic activity of eTkar was better than curative one. Conclusions. The antioxidant properties eTkar was proven to be capable to supress the development of liver fibrosis in which the fibrostatic at the dose of 27,5 mg/kg BW was better than the curative activity one at dose 55 mg/Kg. We conclude that fucoidan from brown seaweed Turbinarria decurren from Lampung has the potential to be developed as a preparation antifibrosis of the liver.
Keywords: Fucoidan, Turbinaria decuren, antioxidant, TGF-1, MMP 13, decorin, liver fibrosis.