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Abstrak :
ABSTRAK
Malaria masih menjadi masalah kesehatan di dunia terutama negara tropis, karena angka kesakitan dan kematiannya yang tinggi. Gejala yang berat sampai kematian akibat malaria dipengaruhi respon imun setiap individu maupun ketepatan pengobatan malaria. Penelitian ini bertujuan untuk mengetahui efek respon imun (TNF-α) mencit terinfeksi Plasmodium berghei yang diberi ekstrak akar pasak bumi sebagai antimalaria. Jenis penelitian ini adalah eksperimental in vivo dengan membagi 5 kelompok perlakuan yang berbeda (kontrol, Plasmodium berghei dan akuades, CMC, Plasmodium berghei dan CMC, Plasmodium berghei dan ekstrak akar pasak bumi). Pemeriksaan tingkat parasitemia menggunakan pemeriksaan darah tipis dan tebal. Hasil pemeriksaan TNF-α menggunakan teknik bead based multiplexing technique didapatkan nilai mean flourescence intensity (MFI) yang digunakan sebagai ukuran kadar TNF-α. Hasil penelitian ini menunjukkan kemampuan ekstrak akar pasak bumi sebagai antimalarial, dengan nilai rerata growth inhibit sebesar 88,93%. Hasil uji korelasi menunjukkan adanya hubungan bermakna antara tingkat parasitemia dengan TNF-α (Uji Spearman, r= - 0,838; p=0.002). Hal ini menunjukkan bahwa ekstrak akar pasak bumi dapat mengaktivasi TNF-α yang bekerja sebagai imunoproteksi. Berdasarkan hasil penelitian ini, dapat disimpulkan bahwa pemberian ekstrak akar pasak bumi meningkatkan ekspresi TNF-α yang berhubungan dengan menurunnya tingkat parasitemia pada mencit yang diinfeksi plasmodium berghei.

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ABSTRACT
Malaria is still the main health problem in the world, mainly in tropical countries since its incidence of illness and death is high. The severe symptoms, which may lead to death, are affected not only by the immune response of each individual but also by the efficacy in the malaria treatment. The purpose of this research is to investigate the effect of immune response (TNF-α) of the Plasmodium berghei infected mice which was treated with the pasak bumi root extract as antimalaria. This was in vivo experimental study in which the experimental animals were divided into five different groups (control, Plasmodium berghei and aquades, CMC, Plasmodium berghei and CMC, Plasmodium berghei and pasak bumi root extract). The level of parasitemia were determineted by using thin and thick blood staining. The bead based multiplexing technique was used in the TNF-α examination in order to obtain mean fluorescence intensity (MFI) which was later used as TNF-α standard. The results of this research showed the potential of the pasak bumi root extract as antimalaria with the mean percentage of growth inhibition was 88.93%. The correlation analysis showed a meaningful relation between the parasitemia level and TNF-α (Spearman test, r= - 0,838; p=0.002). This means that the pasak bumi root extract could activate TNF-α which acts as immune protector. In conclucion, the pasak bumi root extract could enhance the TNF-α expression as shown by the decline of the parasitemia level in the Plasmodium berghei infected mouse

Abstrak :
[ABSTRAK
Tuberkulosis penyakit infeksi yang mematikan terutama di negara berkembang, termasuk Indonesia. Upaya pencegahan dengan vaksinasi BCG yang dapat meningkatkan respon imun masih belum maksimal. Faktor yang mempengaruhi keberhasilan vaksin adalah status imun host, genetik dan kualitas/kuantitas vaksin. Indonesia sebagai negara kaya tanaman obat, misalnya pasak bumi (Eurycoma longifolia Jack) digunakan sebagai antimalaria serta meningkatkan imun tubuh. Penelitian ini menilai efek ekstrak akar pasak bumi sebagai imunomodulator terutama IFN-ɣ, TNF-α dan IL-10 pada mencit yang diberi vaksin BCG. Eksperimental in vivo dan in vitro darah mencit di kultur pada medium RPMI dengan stimulasi PHA dan BCG. Analisis tidak ada perbedaan yang bermakna (p≥0,05) diantara kelompok perlakuan, analisa dari nilai median terlihat adanya efek ekstrak pasak bumi terhadap peningkatan TNF-α, dan tidak berpengaruh terhadap produksi IFN-γ dan IL-10 pada mencit yang divaksin BCG. Ekstrak akar pasak bumi mempengaruhi respon imun tubuh mencit yang diberi vaksin BCG, walau tidak besar maknanya.

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ABSTRACT
Tuberculosis is a deadly infectious disease that occurs mainly in developing countries, including Indonesia. Preventive efforts by BCG vaccination to improve the immune response is still not maximum. Factors that affect the success of vaccine are the host immune system, the host genetic and the quality/quantity of the vaccine. Indonesia is rich in medicinal plants, one of those is Pasak Bumi (Eurycoma longifolia Jack) that is widely used as antimalaria and to improve immunity. The research assessed the effects of extracts of Pasak Bumi roots as immunomodulator by measuring IFN-ɣ, TNF-α and IL-10 on mice that were given with BCG vaccine. In vivo and in vitro experiments of mice blood cultured in RPMI medium stimulated with PHA and BCG. The result has shown no significant difference (p≥0,05) among the treatment group, result of median values has shown the effect of Pasak Bumi extract to an increase of TNF-α, and has no effect on the production of IFN-γ and IL-10 in mice vaccinated BCG. Extract of pasak bumi roots affects the immune response of mice that have got BCG vaccine, although it has no significant meaning., Tuberculosis is a deadly infectious disease that occurs mainly in developing countries, including Indonesia. Preventive efforts by BCG vaccination to improve the immune response is still not maximum. Factors that affect the success of vaccine are the host immune system, the host genetic and the quality/quantity of the vaccine. Indonesia is rich in medicinal plants, one of those is Pasak Bumi (Eurycoma longifolia Jack) that is widely used as antimalaria and to improve immunity. The research assessed the effects of extracts of Pasak Bumi roots as immunomodulator by measuring IFN-ɣ, TNF-α and IL-10 on mice that were given with BCG vaccine. In vivo and in vitro experiments of mice blood cultured in RPMI medium stimulated with PHA and BCG. The result has shown no significant difference (p≥0,05) among the treatment group, result of median values has shown the effect of Pasak Bumi extract to an increase of TNF-α, and has no effect on the production of IFN-γ and IL-10 in mice vaccinated BCG. Extract of pasak bumi roots affects the immune response of mice that have got BCG vaccine, although it has no significant meaning.]

Abstrak :
[ABSTRAK
Kemampuan Mycobacterium tuberculosis yang dapat memanipulasi dan menghindari respon imun adalah tantangan dalam mencari terapi dan vaksinasi efektif. Penelitian ini untuk menguji potensi ekstrak metanol akar Eurycoma longifolia Jack dalam memodulasi peningkatan IgA dan IgG mencit putih jantan yang divaksinasi BCG, sekaligus untuk menganalisis peningkatan sitokin proinflamasi yang berhubungan dengan produksi IgA dan IgG. Plasma diambil sebelum dan setelah vaksinasi BCG pada 18 ekor mencit yang dibagi ke dalam 2 kelompok; perlakuan dan kontrol. Kadar IgA, IgG, TNF-! dan IL-10 diukur dengan metode ELISA. Hasil menunjukkan bahwa peningkatan IgA kelompok air (0,33±0,16) lebih tinggi dibandingkan kelompok pasak bumi (0,30±0,30), sedangkan peningkatan IgG kelompok pasak bumi (0,38±0,25) terlihat lebih tinggi dibandingkan kelompok air (0,29±0,35). Rasio TNF-!/IL-10 kelompok pasak bumi (0,46±0,07) lebih tinggi dibandingkan kelompok air (0,41±0,05). Terdapat korelasi antara TNF-! dengan IgA pada kelompok air (r=0,601, p=0,035) juga dengan IgG pada kelompok pasak bumi (r=0,559, p=0,059). Disimpulkan, pemberian pasak bumi cenderung berpotensi memodulasi peningkatan IgG, tetapi tidak IgA. Selain itu, pemberian pasak bumi juga cenderung meningkatkan sitokin proinflamasi yang mempengaruhi produksi IgA dan IgG.

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ABSTRACT
The ability of Mycobacterium tuberculosis to manipulate and evade host?s immune response is a challenge in finding an effective therapy and vaccination. The purpose of this research were to investigate the potency of methanol extract of the roots of Eurycoma longifolia Jack as immunomodulators for male white mice vaccinated with BCG, as well as to analyze the increase of proinflammatory cytokines associated with IgA and IgG production. Plasma were taken before and after BCG vaccination from 18 mice, that were divided into 2 groups; treatment and control. IgA, IgG, TNF-! and IL-10 levels were measured by using ELISA. The results showed that the enhancement of IgA group with water (0,33±0,16) was higher than group with pasak bumi (0,30±0,30). On the other hand the enhancement of IgG group with pasak bumi (0,38±0,25) was higher than group with water (0,29±0,35). The ratio of TNF-!/IL-10 group with pasak bumi (0.46±0.07) was higher than group with water (0.41±0.05). There are correlarions between TNF-! and IgA in the group with water (r=0.601, p=0.035) as well as between TNF- ! and IgG in group with pasak bumi (r=0.559, p=0.059). As the conclusion, methanol extract of pasak bumi root tend to potentially modulate the increase IgG production, but not IgA. In addition, it also tends to increase the proinflamasi cytokines associated with IgA and IgG production.;The ability of Mycobacterium tuberculosis to manipulate and evade host’s immune response is a challenge in finding an effective therapy and vaccination. The purpose of this research were to investigate the potency of methanol extract of the roots of Eurycoma longifolia Jack as immunomodulators for male white mice vaccinated with BCG, as well as to analyze the increase of proinflammatory cytokines associated with IgA and IgG production. Plasma were taken before and after BCG vaccination from 18 mice, that were divided into 2 groups; treatment and control. IgA, IgG, TNF-! and IL-10 levels were measured by using ELISA. The results showed that the enhancement of IgA group with water (0,33±0,16) was higher than group with pasak bumi (0,30±0,30). On the other hand the enhancement of IgG group with pasak bumi (0,38±0,25) was higher than group with water (0,29±0,35). The ratio of TNF-!/IL-10 group with pasak bumi (0.46±0.07) was higher than group with water (0.41±0.05). There are correlarions between TNF-! and IgA in the group with water (r=0.601, p=0.035) as well as between TNF- ! and IgG in group with pasak bumi (r=0.559, p=0.059). As the conclusion, methanol extract of pasak bumi root tend to potentially modulate the increase IgG production, but not IgA. In addition, it also tends to increase the proinflamasi cytokines associated with IgA and IgG production., The ability of Mycobacterium tuberculosis to manipulate and evade host’s immune response is a challenge in finding an effective therapy and vaccination. The purpose of this research were to investigate the potency of methanol extract of the roots of Eurycoma longifolia Jack as immunomodulators for male white mice vaccinated with BCG, as well as to analyze the increase of proinflammatory cytokines associated with IgA and IgG production. Plasma were taken before and after BCG vaccination from 18 mice, that were divided into 2 groups; treatment and control. IgA, IgG, TNF-! and IL-10 levels were measured by using ELISA. The results showed that the enhancement of IgA group with water (0,33±0,16) was higher than group with pasak bumi (0,30±0,30). On the other hand the enhancement of IgG group with pasak bumi (0,38±0,25) was higher than group with water (0,29±0,35). The ratio of TNF-!/IL-10 group with pasak bumi (0.46±0.07) was higher than group with water (0.41±0.05). There are correlarions between TNF-! and IgA in the group with water (r=0.601, p=0.035) as well as between TNF- ! and IgG in group with pasak bumi (r=0.559, p=0.059). As the conclusion, methanol extract of pasak bumi root tend to potentially modulate the increase IgG production, but not IgA. In addition, it also tends to increase the proinflamasi cytokines associated with IgA and IgG production.]

Abstrak :
Natural Deep Eutectic Solvents (NADES) semakin banyak menarik perhatian sebagai pelarut alternatif ramah lingkungan pengganti pelarut organik konvensional yang toksik dan berbahaya bagi lingkungan. NADES tidak volatil, memiliki solubilitas tinggi, toksisitas rendah, dan selektivitas yang dapat diatur. Pada penelitian ini, ekstraksi dilakukan dengan metode NADES-UAE dengan biomarker yaitu eurikumanon. Eurikumanon merupakan senyawa utama yang terdapat pada tanaman akar pasak bumi dan memiliki efek farmakologis. Sebagai metode pembanding dilakukan ekstraksi refluks dengan pelarut metanol. Adapun NADES yang dipakai merupakan campuran antara kolin klorida sebagai akseptor ikatan hidrogen dengan tiga gula poliol sebagai donor ikatan hidrogen yaitu gliserol, xilitol, dan sorbitol. Optimasi metode ekstraksi dilakukan menggunakan Response Surface Methodology (RSM). Faktor yang dioptimasi untuk kondisi ekstraksi meliputi waktu ekstraksi dan persen penambahan air. Kadar eurikumanon diukur menggunakan Kromatografi Cair Kinerja Tinggi (KCKT) dengan fase gerak asam format 0,1% : asetonitril (90:10 v/v), laju alir 1,0 mL/menit, dan panjang gelombang 254 nm. Kolin Klorida-gliserol merupakan NADES terbaik untuk mengekstraksi eurikumanon dibandingkan kolin klorida-xilitol, dan kolin klorida-sorbitol. Kadar eurikumanon tertinggi diperoleh yaitu sebesar 16,90 mg/g pada waktu ekstraksi 50 menit dan penambahan air 50%. Hasil penetapan kadar eurikumanon dengan ekstraksi NADES-UAE yaitu sebesar 16,90 mg/g berbeda signifikan dengan hasil penetapan kadar dengan ekstraksi refluks sebesar 0,1002 mg/g. Berdasarkan penelitian ini dapat disimpulkan bahwa NADES kolin klorida-gliserol dapat menarik senyawa eurikumanon dari akar pasak bumi.

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Natural Deep Eutectic Solvents (NADES) are increasingly attracting attention as an environmentally friendly alternative to conventional organic solvents that are toxic and harmful to the environment. NADES is not volatile, has high solubility, low toxicity, and adjustable selectivity. In this study, extraction was carried out by the NADES-UAE method with a biomarker, namely eurycomanone. Eurycomanone is the main compound found in pasak Bumi roots and has many pharmacological effects. As a comparison method, reflux extraction uses metanol as a solvent. The NADES used is a mixture of choline chloride as a hydrogen bond acceptor with three polyols sugar as hydrogen bond donors, namely glycerol, xilitol, and sorbitol. The optimization of the extraction method uses the Response Surface Methodology (RSM). Optimized factors for extraction conditions include extraction time and percents water addition. Eurycomanone levels were measured using High Performances Liquid Chromatography (HPLC) with a 0.1% formic acid phase: acetonitrile (90:10 v / v), a flow rate of 1.0 mL/min, and a wavelength of 254 nm. Choline Chloride-glycerol is the best NADES for extracting eurycomanone. Optimal conditions for obtaining the highest eurycomanone levels are at 50 minutes extraction time and 50% water addition with levels of 16,90 mg/g. The results of the determination of eurycomanone levels by NADES-UAE extraction 16,90 mg/g were significantly different from reflux extraction 0.1002 mg/g. This research concludes that NADES choline chloride-glycerol can attract eurycomanone compounds from the pasak Bumi roots.

Abstrak :
Natural Deep Eutectic Solvents (NADES) telah dikembangkan sebagai pelarut alternatif dari pelarut organik karena memiliki sifat tidak toksik, ramah lingkungan, biodegradable, preparasi mudah, dan harga terjangkau. Senyawa eurikumanon merupakan kandungan tertinggi yang ditemukan di akar tanaman pasak bumi (Eurycoma Longifolia Jack) yang memiliki banyak kegunaan seperti pengobatan aprodisiak, antimalarial, antioksidan dan lainnya. Pada penelitian ini ekstraksi senyawa eurikumanon dari akar pasak bumi dilakukan dengan metode Ultrasound-Assisted Extraction menggunakan NADES kombinasi kolin klorida-Gula (glukosa, fruktosa, dan sukrosa). Optimasi metode ekstraksi dilakukan menggunakan Response Surface Methodology (RSM). Faktor yang dioptimasi untuk kondisi ekstraksi meliputi persentase penambahan air dan waktu ekstraksi. Analisis kadar senyawa eurikumanon dilakukan dengan menggunakan Kromatografi Cair Kinerja Tinggi (KCKT) dengan fase gerak larutan asetonitril:asam format 0,01% (10:90 v/v), laju alir 1,0 mL/menit dan dideteksi pada panjang gelombang 254 nm. Kolin klorida-glukosa merupakan NADES terbaik untuk mengekstraksi eurikumanon dibandingkan dengan jenis NADES lainnya. Kondisi optimal untuk memperoleh eurikumanon dengan kadar tertinggi yaitu pada penambahan air 60% dan waktu ekstraksi 60 menit dengan kadar eurikumanon sebesar 12,28 mg/g. Berdasarkan penelitian ini, dapat disimpulkan bahwa NADES kolin klorida-gula dapat menarik senyawa eurikumanon dari akar pasak bumi. Hasil ekstraksi yang diperoleh dibandingkan dengan ekstraksi menggunakan pelarut etanol 96% dengan metode sokhletasi sebesar 0,23 mg/g. Kadar senyawa eurikumanon yang diperoleh dengan metode UAE-NADES lebih tinggi.

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Natural Deep Eutectic Solvents (NADES) has been developed as an alternative to organic solvents because they are non-toxic, environment-friendly, biodegradable, easy to produce, and have a low cost. Eurycomanone is the highest compound found in pasak bumi roots (Eurycoma Longifolia Jack) that has many pharmacological effects susch as antimalarial, antioxidant, and aphrodisiac. In this research, the extraction of eurycomanone compounds from the pasak bumi roots was carried out by Ultrasound-Assisted Extraction method using NADES of choline chloride-sugar (glucose, fructose, and sucrose) based. The optimization of the extraction method was done using Response Surface Methodology (RSM). Optimized factors for extraction conditions include the water percentage and extraction time. The analysis of eurycomanone compound was performed using High Performance Liquid Chromatography (HPLC) with a mobile phase of acetonitrile:0,01% formic acid (10:90 v/v), the flow rate of 1,0 mL/min and detected at a wavelength of 254 nm. Choline chloride-glucose showed the highest result for extracting eurycomanone compared to other types of NADES. The optimal conditions are obtained at 60 minutes of extraction time, and 60% percent of water gives the highest level of eurycomanone with 12,28 mg/g. Based on this research, it can be concluded that NADES choline chloride-sugar could attract eurycomanone compounds from the pasak bumi roots. The extraction results were compared to extraction using soxhletation methods with 96% ethanol, the level of eurycomanone detected at 0,23 mg/g. NADES solvents produced higher levels of eurycomanone compounds.