Mekanisme Kerja Protektif Ekstrak Andrographis paniculata pada Mencit Balb/c Selama Infeksi Plasmodium berghei ANKA Kajian Kapasitas Produksi Interferon-gamma Kajian Kapasitas Produksi Interleukin-12

Abstract

Background: A. paniculata are able to modulate immune responses. IFN-y and IL-12 is a protective cytokine during Plasmodium infection. However it is not known whether A. paniculata herbs has an effect on IFN-y and IL-12 production capacity in Balb/c mice inoculated by P. berghei ANKA. IL-12 upregulate IFN-y produstion. Objective: to determine an increase and a correlation of IFN-y and IL-12 production capacity in Balb/c mice inuculated by P. berghei ANKA and treated with A. paniculata. Methods: This study was pure experimental design with post test only control group design. Total Balb/c mice used were 36, which devided equally in treated and control groups. Treated group were receive A. paniculata herbs, while control group receive CMC-Na 0,5%, a herb diluent orally since 7 days before inoculation and p i P. berghei ANKA. Both groups were inoculated by 104 erythrocyte parasitized by P. berghei ANKA. Sample were spleenocytes of Balb/c mice p.i P. berghei ANKA. Spleenocytes and peritoneal macrophages were isolated on day 3, 6, and 8 p.i and were in vitro stimulated by PHA or LPS, a T cell mitogen and a macrophages mitogen, repectively. IFN-y and IL-12 production capacity were measured from cell culture supernatant by using Elisa method. Results: Treated group produced a significant higher IFN-y concentration than control group on day 3 p.i of P. berghei ANKA. Statistical analysis between day p.i. showed that maximum IFN-y concentration in treated and control groups occured on day 3 and 6 p.i, respectively. IL-12 p70 production in treated group were as high as those observed in control. No different of IL-12 p70 production among days observed p.i. P. berghei ANKA in treated and control groups. No correlation were found between IFN-y and IL-12 produced by spleenocytes in both treated and control group. Conclussion: A. paniculata herbs given orally increase IFN-y spleenocytes production capacity on day 3 when IFN-y spleenocytes maximum production p.i of P. berghei ANKA occured. However A. paniculata do not increase IL-12 p70 production p.i of P. berghei ANKA. IL-12 p70 do not correlate with IFN-y production capacity of spleenocytes obtain from Balb/c mice p.i of P. berghei ANKA. Latar Belakang: IFN-y dan IL-12 bersifat protektif selama infeksi plasmodium. A. paniculata mampu menekan perkembangan Plasmodium sekaligus memodulasi respon imun. Belum diketahui bagaimana kapasitas produksi IFN-y dan IL-12 mencit Balb/c yang diberi A. paniculata peroral dan diinokulasi P. berghei ANKA . Tujuan: membuktikan adanya peningkatan dan korelasi kapasitas produksi IFN-y dan IL-12 p.i. P. berghei ANKA pada mencit Balb/c yang diberi A. paniculata. Methode: Penelitian ini merupakan penelitian eksperimental murni dengan desain post test only control group. Jumlah sampel seluruhnya 36 ekor mencit yang dibagi 2 sama banyak secara random menjadi masing-masing kelompok perlakuan dan kontrol.'Kelompok perlakuan menerima herbal A. paniculata, sedangkan kontrol menerima CMC-Na 0,5%, pelarut herbal sejak 7 hari sebelum dan 8 hari pasca.inokulasi P. berghei ANKA. Kedua kelompok diinokulasi 104 eritrosit berparasit P. berghei ANKA. Makrofag peritoneal dan splenosit diisolasi pada hari 3, 6, 8 p.i mencit Balb/c dan secara in vitro diberi stimulasi PHA, atau LPS, masing¬masing mitogen sel T dan makrofag. Kapasitas produksi IFN-y dan IL-12 p70 diukur dari supernatan kultur, dengan metode Elisa. Hasil: Uji beda antara kelompok perlakuan dibanding kontrol memperlihatkan pada hari ke 3 p.i P. berghei ANKA kapasitas produksi IFN-y sel limpa kelompok perlakuan yang diberi stimulasi PHA in vitro lebih tinggi secara bermakna dibanding kontrol. Uji beda antar hari pengamatan menunjukan bahwa kapasitas maksimal produksi IFN-y sel limpa yang diberi stimulasi PHA in vitro kelompok perlakuan dan kelompok kontrol masing-masing didapatkan pada hari ke 3 dan 6 p.i P. berghei ANKA. Perbedaan kapasitas produksi IL-12 p70 tidak ditemukan antara kelompok perlakuan dan kontrol. Perbedaan kapasitas produksi IL-12 p70 antar hari pengamatan tidak ditemukan baik pada kelompok perlakuan maupun kontrol. Kesimpulan: Pada hari ke 3 p.i P. berghei ANKA didapatkan peningkatan produksi IFN-y sel limpa pada mencit Balb/c yang diberi herbal A. paniculata. Sebaliknya peningkatan produksi IL-12 p70 p.i P. berghei ANKA tidak ditemukan pada mencit yang diberi herbal A. paniculata. IL-12 p70 tidak berkorelasi dengan kapasitas produksi IFN-y sel limpa mencit Balb/c p.i P. berghei ANKA