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"Latar belakang: Saat ini prevalens alergi semakin meningkat. Terapi alergi lebih terfokus untuk mengatasi gejala simptomatik. Pendekatan lain adalah terapi imunomodulasi dengan agonis toll like receptor (TLR) 9 yang dapat mengalihkan respon imun sel TCD4+ Th2 ke arah Th1. CpG ODN 2006x3_PD adalah oligonukleotida sintetik kelas B merupakan agonis TLR9 yang berpotensi aman karena mengandung fosfodiester. Pada penelitian ini CpG ODN 2006x3_PD diuji kemampuan untuk mengatasi alergi dengan menggunakan penghantar nanopartikel kitosan, melalui uji in vitro pada sel mononukleus darah tepi dan in vivo pada hewan coba mencit alergi. Metode: Preparasi nanopartikel kitosan dilakukan dengan metode gelasi ion, melalui reaksi ikat silang kitosan dan tripolifosfat. Nanopartikel dikarakterisasi dengan Dynamic light scattering (DLS), zeta sizer dan transmission electron microscope (TEM). Uji pengikatan dilakukan dengan elektroforesis pada gel agarosa 12%, uji toksisitas dan kemampuan aktivasi NF-κB dilakukan pada sel RAW Blue, dengan menggunakan cel counting kit 8 dan kit Quanti blue. Sel mononukleus darah tepi yang distimulasi selama 7 hari pada uji in vitro dan plasma hewan coba mencit Balb/c pada uji in vivo diukur konsentrasi IFNγ, IL-10, IL-13 dan IgE dengan metode ELISA. Hasil: Diperoleh nanopartikel dengan ukuran <300 nm, muatan permukaan positif, bentuk sferis, tidak toksik dan dapat mengaktivasi NF-κB. Uji in vitro pada sel mononukleus darah tepi menunjukkan CpG ODN 2006x3_PD yang dihantarkan nanopartikel kitosan dapat menstimulasi sitokin tipe Th1 IFNγ dan T reg IL-10, menurunkan sitokin tipe Th2 IL-13 namun belum dapat menurunkan secara bermakna produksi IgE total pada sel mononukleus darah tepi individu sehat dan alergi Aplikasi intra nasal 10 ug/kali, 3 kali seminggu selama 3 minggu CpG ODN 2006x3_PD dan CpG ODN 2006x3_PD yang dihantarkan nanopartikel kitosan pada mencit Balb/c yang diinduksi alergi dengan ovalbumin dapat menstimulasi sitokin tipe Th1 IFNγ dan Treg IL-10, namun belum dapat menurunkan secara bermakna sitokin tipe Th2 IL-13 pada plasma mencit. Aplikasi CpG ODN 2006x3_PD dapat menurunkan produksi IgE spesifik anti ovalbumin pada plasma mencit meskipun belum dapat menurunkan produksi IgE total. Simpulan: CpG ODN 2006x_PD dapat menjadi kandidat imunostimulator yang potensial pada alergi.

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Background: Currently, prevalence of allergy has increased in worldwide. Allergy treatment is mainly focused to reduce clinical symptoms. Another approach is immunomodulation therapy which utilizes toll like receptor (TLR) 9 agonist that may redirect pro-allergenic Th2 biased CD4+ T cell response toward Th1. CpG ODN 2006x3_PD which is classified as synthetic oligonucleotides B has potential as a safe TLR9 agonist due to its natural backbone. In this study, CpG ODN 2006x3_PD was examined about its ability in overcoming allergies by using chitosan nanoparticles delivery, through in vitro tests on peripheral blood mononuclear cells and in vivo through allergic mice animal model. Methods: Chitosan nanoparticles was prepared by ionic gelation method, through crosslinking reaction of chitosan and tripolyphosphate. Nanoparticles are characterized by Dynamic light scattering (DLS), zeta sizer and transmission electron microscope (TEM). The binding test was carried out with electrophoresis on 12% agarose gel, toxicity test and NF-κB activation ability performed on RAW Blue cells, using cel counting kit 8 and Quanti blue kit. IFNγ, IL-10, IL-13 and IgE level of in vitro tests of peripheral blood mononuclear cells after 7 days stimulation and Balb/c mice plasma of in vivo study were measured by ELISA method.Results: Less than 300 nm, positive surface charge, spherical shape and nontoxic chitosan nanoparticles were obtained. These nanoparticles could deliver CpG ODN to activate NF-κB of mouse RAW-Blue cells effectively. In vitro assays of peripheral blood mononuclear cells showed that CpG ODN 2006x3_PD delivered by chitosan nanoparticles may stimulate Th1 IFNγ and T reg type cytokines IL-10, also decrease the Th2-type cytokine IL-13 but it couldn't inhibit total IgE production in peripheral blood mononuclear cells significantly. Intranasal application of 10 ug, 3 times per week for 3 weeks of CpG ODN 2006x3_PD and CpG ODN 2006x3_PD which were delivered by chitosan nanoparticles in allergen induced Balb/c mice could stimulate Th1 IFNγ and Treg type cytokines IL-10, but it couldn't significantly reduce the Th2-type cytokine IL-13 in mice plasma . The CpG ODN application decreased the specific IgE production of anti ovalbumin in mice plasma although it couldn't significantly reduce total IgE production. Conclusions: CpG ODN 2006x_PD could be a potential candidate for allergic immunostimulator."

"Komplikasi penyakit perlemakan hati non-alkoholik (PHNA) ditemukan pada 67% populasi memenuhi kriteria sindrom metabolik. Acalypha indica L. (AI) adalah herbal yang telah diketahui memiliki efek anti-oksidan, dan anti-inflamasi. Penelitian ini bertujuan membuktikan efek AI terhadap mekanisme pertahanan imun yang dibawa. Penelitian dilakukan dengan molecular docking terhadap senyawa AI pada TLR9, NFκB, TNFα, dan perubahan histopatologik hati. Model hewan steatohepatitis pada tikus Sprague-Dawley didapat dari induksi diet tinggi fruktosa, dan kolesterol (DTFK) selama 12 minggu. Terapi diberikan selama 8 minggu. Dua puluh lima tikus dibagi ke dalam 5 kelompok: Normal (K1), DTFK (K2), DTFK+AI, 400 mg (K3), kombinasi AI, 400 mg +gemfibrozil (Gem) 31 mg (K4) dan Gem 31 mg (K5) masing-masing per kgBB. Molecular docking untuk mengidentifikasi interaksi antara molekul hidrogen senyawa AI dengan residu asam amino TLR9, NFκB, TNFα. Perubahan morfologi hati dinilai dengan cara skoring. Analisis statistik yang dilakukan adalah uji Kruskall Wallis post hoc Mann Whitney, dilanjutkan dengan uji korelasi Spearman. Hasil molecular docking menunjukkan, selain senyawa flavonoid, ditemukan senyawa alkaloid beta-sitosterol, dan stigmasterol yang dapat berikatan dengan ketiga marker inflamasi dengan nilai binding energy terbaik. Senyawa lain dasycarpidan-1-methanol, acetate (ester), fenofibrate, quinine. Pemberian AI menurunkan hipertrofi (p=0,031), steatosis makrovesikular (p=0,018), fokus inflamasi (p=0,005). Pemberian AI juga menurunkan ekspresi TLR9 (p=0,009), NFκB (p=0,009), TNFα (p=0,009), akan tetapi tidak sebaik pemberian kombinasi AI+Gem.

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Complications of non-alcoholic fatty liver disease (NAFLD) include 67% of the criteria for metabolic syndrome. Acalypha indica L., (AI) which is one of a herbal plant had been known as anti-oxidant and anti-inflammatory effects. The effect of AI for therapy investigated by looking of the immune defense mechanisms. This researched was assessed by molecular docking approached on TLR9, NFκB, TNFα expression and liver morphological changes. Animal models of steatohepatitis were collected from high- fructose and cholesterol diet (HFCD) of Sprague-Dawley rats for 12 weeks and followed by therapy for 8 weeks. There were 5 groups from twenty five researched rats, include normal group (K1), HFCD group (K2), HFCD group supplemented with 400 mg Acalypha indica L. (K3), combination between 400 mg AI.+gemfibrozil (Gem) 31 mg (K4) and Gem 31 mg/kg (K5) in kgBW, respectively. The results of molecular docking were carried out by assessing the interaction between hydrogen molecules of AI compounds and amino acid residues in TLR9, NFκB, TNFα. Morphological changes were assessed by scoring system. Statistical analyzed used Kruskall Wallis with post hoc Mann Whitney test continued by Spearman correlation test. The molecular docking analysis showed that, an alkaloid compounds were found besides the flavonoid compounds that can bind to the binding pocket of inflammatory markers with the best binding energies. Other compounds, there are dasycarpidan-1-methanol, acetate (ester), fenofibrate and quinine. Supplementation of AI would reduced hypertrophy (p=0.031), macrovesicular steatosis (p=0.018), inflammation foci (p=0.005) and also decreased of TLR9 (p=0.009), NFκB (p=0.009), TNFα (p=0.009) expression, but not as good as the combination of AI+Gem."