EFEK IMUNOMODULATOR DUA SENYAWA NEOLIGNAN HASIL ISOLASI DARI EKSTRAK METANOL DAUN SIRIH MERAH (Piper crocatum Ruiz & Pav.): Kajian Imunitas Seluler dan Humoral

Yustina Sri Hartini, Prof. Dr. Subagus Wahyuono, M.Sc., Apt.

2014 | Disertasi | S3 Ilmu Farmasi

Daun sirih merah (Piper crocatum Ruiz & Pav.) banyak digunakan sebagai obat tradisional. Beberapa aktivitas farmakologis ekstrak dari daun sirih merah telah dilaporkan. Secara in vivo, ekstrak daun sirih merah meningkatkan aktivitas fagositosis makrofag tetapi tidak berpengaruh terhadap proliferasi limfosit maupun titer imunoglobulin G. Uji in vitro senyawa hasil isolasi (1 dan 2) dari daun sirih merah menunjukkan bahwa 1 dan 2 pada dosis 5μg/ml dapat meningkatkan aktivitas fagositosis makrofag. Isolat 1 dan 2 diidentifikasi sebagai neolignan yakni senyawa 2-allyl-4-(1'-hydroxy-1'-(3 \\", 4\\", 5 \\"-trimethoxyphenyl) propan-2'-yl) -3,5- dimethoxycyclohexa-3, 5-dienone dan senyawa 2-allyl-4-(1'-acetyl-1'-(3 \\", 4\\", 5 \\"- trimethoxyphenyl) propan-2'-yl) -3,5-dimethoxycyclohexa- 3,5-dienone). Penelitian ini bertujuan menetapkan efek imunomodulator dan efek terhadap gambaran histopatologis dari perlakuan dengan 1 dan 2 secara in vivo pada mencit. Isolat 1 dan 2 (2,5; 5; dan 10 mg/kgBB) diuji efek imunomodulatornya terhadap respon imun mencit BALB/c yang diinduksi dengan Listeria monocytogenes. Efek 1 dan 2 terhadap respon imun seluler ditetapkan melalui uji aktivitas fagositosis makrofag, produksi nitrit oksid, dan proliferasi limfosit; sedangkan efek terhadap respon imun humoral ditetapkan melalui pengukuran titer imunoglobulin G. Lebih lanjut ditetapkan efek 1 dan 2 terhadap produksi interleukin 12 dan interleukin 10. Efek perlakuan 1 dan 2 terhadap gambaran histopatologis hati dan ginjal mencit diperiksa secara mikroskopis dengan pengecatan hematoksilin-eosin. Analisis statistik data efek imunomodulator menggunakan one-way ANOVA dilanjutkan uji Tukey, sedangkan analisis gambaran histopatologis hati dan ginjal secara deskriptif. Hasil penelitian menunjukkan bahwa secara in vivo, 1 dan 2 (5 dan 10 mg/kg berat badan) meningkatkan aktivitas fagositosis makrofag. Pada dosis 2,5; 5; dan 10 mg/kg berat badan, 1 dan 2 meningkatkan produksi nitrit oksid dan interleukin 12, tetapi pengaruh terhadap proliferasi limfosit, titer imunoglobulin G dan produksi interleukin 10 tidak berbeda bermakna dengan kontrol. Perlakuan dengan 1 (2,5; 5; dan 10 mg/kg berat badan) tidak berpengaruh terhadap gambaran histopatologis hati dan ginjal, sedangkan 2 pada dosis 2,5 mg/kg berat badan menyebabkan degenerasi hidropis dan dosis 5 mg/kg berat badan menyebabkan nekrosis hati.

Red betel (Piper crocatum) leaf is widely used as a traditional medicine in Indonesia. Some of the pharmacological activities of red betel leaf extract have been reported. The in vivo study reported that red betel leaf extract had macrophage phagocytic activity but had no significantly effect on lymphocyte proliferation and immunoglobulin G titre. In vitro study of two isolated compounds (1 and 2) from red betel leaf extract showed that at the dose of 5 μg/ml increased macrophage phagocytic activity. The 1 dan 2 were identified as neolignans, i.e., 2-allyl-4-(1'- hydroxy-1'-(3 \\", 4\\", 5 \\"-trimethoxyphenyl) propan-2'-yl) -3,5-dimethoxycyclohexa- 3, 5-dienone and 2-allyl-4-(1'-acetyl-1'-(3 \\", 4\\", 5 \\"-trimethoxyphenyl) propan-2'- yl) -3,5-dimethoxycyclohexa- 3,5-dienone. This study aim to determine the in vivo immunomodulatory and histopathological features effect of 1 dan 2 on mice. Isolate 1 and 2 were tested on BALB/c mice immune response induced by Listeria monocytogenes. The 1 and 2 cellular immune response determined through macrophage phagocytic activity test, nitric oxide production, and lymphocyte proliferation, whereas the compounds effect on the humoral immune response determined by measuring imunoglobulin G titre. Furthermore, 1 and 2 effect on the IL-12 and IL-10 productions were also determined. Histopathological features effects of 1 and 2 were observed from the liver and kidney which made by hematoxyllin-eosin staining, and observed microscopiccally. The data of immunomodulatory effects were analysed using one-way ANOVA followed by Tukey test, while the liver and kidney histopathological features were analysed descriptively. This in vivo study show that 1 and 2 (5 mg/kg body weight) increase the phagocytic activity of macrophages. At the doses of 2.5, 5, and 10 mg/kg body weight, 1 and 2 increase the production of nitric oxide and interleukin 12 but the mice lymphocyte proliferation, immunoglobulin G titre, and interleukin 10 production have no significantly differences compare to control. The treatment with 1 (2.5, 5, and 10 mg/kg body weight) has no histopathological features effect on both liver and kidney, but at the dose of 2.5 and 5 mg/kg body weight 2 induce liver hidropic degeneration and necrosis respectively.

Kata Kunci : Piper crocatum Ruiz & Pav., isolasi, imunomodulator, in vivo, gambaran histopatologis