EFEK IMUNOMODULATOR INFUSA KULIT BATANG MASOYI (Massoia aromatica Becc.)

HARLYANTI M MASHAR, Dr.rer.nat. Triana Hertiani, M.Si., Apt.;Dr. Ag. Yuswanto, SU., Apt.

2016 | Tesis | S2 Ilmu Farmasi

Secara tradisional, kulit batang masoyi oleh masyarakat Papua digunakan sebagai obat tradisional untuk mengobati demam dan radang dengan cara direbus atau dikukus. Minyak atsirinya telah terbukti berpotensi sebagai imunomodulator secara in vitro, tetapi belum ada bukti ilmiah mengenai efek imunomodulatornya secara in vivo. Penelitian ini bertujuan untuk mengkaji potensi infusa kulit batang masoyi sebagai agen imunomodulator dengan mengevaluasi efeknya terhadap tikus Wistar secara in vitro dan in vivo. Pengujian secara in vitro dilakukan menggunakan ekstrak air. Ekstrak air diperoleh dengan membuat infusa 10%, kemudian di freeze dried. Kadar C-10 massoialactone ditentukan dengan metode KLT-densitometri. Ekstrak air diuji dengan konsentrasi 2,5; 5; 10; 20; dan 40 µg/ml media. Kemampuan sel dalam memfagosit lateks ditentukan dalam indeks fagositosis (IF) melalui 300 makrofag. Uji proliferasi limfosit dilakukan dengan metode reduksi MTT dan dianalisis menggunakan pembaca mikroplate pada panjang gelombang 595 nm. Uji in vivo dilakukan menggunakan infusa 20%. Tikus Wistar dibagi menjadi 5 kelompok, yaitu baseline, Dox 10 mg/kg (i.p), 3 kelompok perlakuan dengan dosis 100, 300, dan 500 mg/100 g BB (p.o). Infusa diberikan setiap hari selama 28 hari, Dox diberikan pada hari ke-1 dan 14. Sampling darah dilakukan pada hari ke-0, 14, dan 28. Penentuan profil sel limfosit TCD4+ dan TCD8+ dilakukan menggunakan flow cytometer. Kadar C-10 massoialactone dalam infusa konsentrasi 10 dan 20% (b/v) masing-masing adalah 0,29 dan 0,70 mg/ml. Hasil uji in vitro menunjukkan bahwa ekstrak air secara signifikan meningkatkan aktivitas fagositosis makrofag dibandingkan dengan kontrol. Variasi konsentrasi menunjukkan adanya korelasi dengan peningkatan IF sel makrofag. Konsentrasi 40 µg/ml mununjukkan aktivitas yang paling tinggi yaitu 70,51±1,11%. Uji proliferasi sel limfosit secara signifikan meningkat, tetapi nilai IS<2 menunjukkan tidak memberikan efek terhadap limfosit. Hasil dari uji in vivo menunjukkan penurunan jumlah sel TCD4+ dan TCD8+ pada kelompok perlakuan dibanding baseline. Rasio TCD4+/TCD8+ tidak berbeda signifikan dibanding kelompok baseline. Penelitian ini menunjukkan bahwa kulit batang masoyi memiliki kemampuan dalam meningkatkan aktivitas fagositosis sel makrofag, tetapi tidak memberikan efek terhadap sel limfosit.

Traditionally, massoia bark is used by local people in Papua as traditional medicine againt fever and inflamation by boiling or steaming. The massoia essential oil has been proven to be a potent immunomodulator in vitro, but no scientific proof about its immunomodulatory effect in vivo. This study aimed to investigate the potency of massoia bark as immunomodulatory agent by evaluating its effects on Wistar rat by in vitro and in vivo assay. Water extract was used for in vitro assay. Water extract was obtained by making decoction 10%, followed by freeze dried. The C-10 massoialactone content were determined by TLC-densitometry method. In vitro assay was performed in a concentration of 2.5; 5; 10; 20; and 40 µg/ml media. The capability of macrophages which phagocyted the latex beads was evaluated by phagositic index through 300 macrophages. The effect on lymphocytes proliferation was observed by the MTT reduction method and analyzed using microplate reader at 595 nm. Decoction 20% was used for in vivo assay. Wistar rats was divided into 5 groups, i.e. baseline, Dox 10 mg/kg (i.p), 3 treatment groups with dose of 100, 300, and 500 mg/100 g BW (p.o). Decoction were given every day for 28 days, Dox was given on days 1 and 14. Blood samples were taken at days 0, 14 and 28 through the eye orbital vein. Determination of the TCD4+and TCD8+ lymphocyte cells amounts were done by using the Flow Cytometer. The content of C-10 massoialactone on massoia bark decoction with concentration of 10% and 20% was 0.29 dan 0.70 mg/ml, respectively. Result from the in vitro assay showed that the water extract significantly increased the macrophage phagocytosis activity in comparison to control. The concentration variation showed dependant manner with the macrophage phagocytic index enhancement. Concentration of 40 µg/ml showed the highest activity i.e. 70.51±1.11%. The lymphocytes proliferation was significantly increased, but IS value < 2 suggesting no effect on lymphocytes. Result from the in vivo assay showed the decreased of TCD4+ and TCD8+ cells amount on treatment groups compared to baseline. TCD4+/TCD8+ ratio was not significantly different compared to baseline. This study showed that massoia bark extract increased the macrophage phagocytosis activity, but on lymphocyte cell proliferation activity.

Kata Kunci : Imunomodulator, kulit batang masoyi, fagositosis sel makrofag, doksorubisin, TCD4+ dan TCD8+