Albab, FatmaAyatiliulil (2012) Modulasi Sistem Immun Lokal dan Sistemik pada Mencit (Mus musculus) BALB/c Pasca Paparan Asap Kretek Berfilter Divine. Magister thesis, Universitas Brawijaya.
Abstract
Filter Divine merupakan salah satu aplikasi sains nanomaterial pada bidang medis sebagai alternatif solusi permasalahan toksisitas rokok dan terapi. Filter Divine mampu memediasi transformasi asap rokok sehingga dihasilkan partikel asap dengan ukuran nanometer dan bersifat sebagai scavenger. Transformasi dapat memberikan perbedaan karakter fisikokimiawi dan mempengaruhi karakter biologis nanopartikel yang dihasilkan, sehingga perlu dilakukan karakterisasi interaksinya dengan sistem biologis. Secara umum, nanopartikel diketahui dapat memodulasi sitem immun, baik pada rute administrasi lokal utama, maupun secara sistemik akibat kemampuan translokasinya menuju sirkulasi limfatik dan darah. Akan tetapi, studi interaksi nanopartikel dengan sistem immun pada penelitian yang ada, menunjukkan modulasi yang inkonsisten dan masing-masing jenis nanopartikel memiliki tipe modulasi yang berbeda. Interaksi tersebut perlu diketahui sehingga didapatkan immunomodulasi yang tepat dan menghindari immunotoksisitas. Data terkait hal tersebut pada aplikasi filter Divine pada rokok tipe kretek masih belum tersedia. Dengan demikian, perlu dilakukan penelitian untuk mengetahui apakah ada perbedaan modulasi sistem immun innate dan sistem immun adaptif pada paru (lokal), spleen dan darah perifer (sistemik) yang meliputi modulasi kuantitatif dan kualitatif serata potensinya dalam menimbulkan alterasi histologis paru pada mencit pasca paparan asap rokok tipe kretek berfilter Divine, sehingga dapat diberikan gambaran interaksinya secara holistik. Penelitian eksperimental ini dilakukan menggunakan Rancangan Acak Lengkap dengan 6 ulangan. Mencit BALB/c jantan dibagi atas kelompok tanpa paparan (kontrol positif) dan kelompok dengan paparan asap rokok kretek (33 mg Tar dan 2.3 mg Nikotin) berfilter biasa (kontrol negatif), berfilter Divine : B, DE dan DF. Dosis paparan yang digunakan adalah asap mainstream dari 4 batang rokok/hari (pagi-sore) selama 3 hari dan dipaparkan secara whole body exposure dengan bantuan smoking pump berkecepatan 0.23 m/s. Sehari pasca paparan terakhir dilakukan pengambilan darah perifer untuk analisis darah lengkap. Dilakukan sectio pada mencit untuk mengambil organ paru dan spleen . Paru lobus kiri difiksasi dengan PFA 4% untuk dibuat preparat HE dan dilakukan analisis histologi sesuai metode D`hulst et al. (2005). Dilakukan analisis flowcytometry pada paru lobus kanan serta spleen pada populasi sel Gr-1 + , T CD4 + , T CD8 + , B220 + , CD62L + dan sel Treg CD4 + CD25 + . Uji kualitatif dilakukan dengan uji aktivasi limfosit dari organ spleen melalui uji ekspresi sitokin secara in vitro. Data kuantitatif dianalisis dengan uji one-way ANOVA (p 0.05) dengan uji lanjut Tukey HSD menggunakan SPSS 16.0 for Windows. Dilakukan analisis cluster
English Abstract
Divine Filter is one of nanomaterial science application as alternative solution towards cigarette toxicity and medical rapy. Special filters are applied and mediating smoke transformation and generate nanometer smoke particulate which has scavenger ability. This transformation leads into different physicochemical properties and affecting biological properties of generated nanoparticles, so that, its interaction with biological system need to be characterized. Nanoparticle commonly has both local and systemic immunomodulation capability due to its ability to translocate into lymphatic and blood circulations. Recent studies indicated that each nanoparticle type has inconsistent immunomodulation and specific individual modulation character. This interaction is important to date, in order to gain desirable immunomodulation and avoid immunotoxicity. Data on application of Divine Filter on kretek cigarette in regarding this issues is not available, so that, a research to examine modulation of innate and adaptive immunity on lung as local main route of exposures, spleen and blood as systemic route with quantitative and qualitative variables, followed by histological examination would be valuable to observe holistic description of interaction. This research was performed in completely randomized design (n=6). 8-weeksold, male Mus musculus BALB/c divided into group with sham exposure (+ control), and 4 o r groups receiving a commercial local kretek (33mg Tar and 2.3mg Nicotine) smoke filtered by common filter (- control), and three Divine filters: B, DE and DF. Whole body exposures were conducted twice a day with two cigarettes in each, for three consecutive days using mainstream smoke generated by smoking pump (V=0.23m/s). 24 hours after final exposures, peripheral blood were drawn from retroorbital sinuses and objected to complete haematological count. Left lobe of lung was fixed with 4% PFA and paraffin embedded. Tissue sections were objected to HE staining and histological examination was performed by D`hulst et al. (2005) method. Right lobe of lung and spleen were objected to flowcytometry analysis of Gr-1 + , T CD4 + , T CD8 + , B220 + , CD62L + and Treg (CD4 + CD25 + ) populations. Functional analysis (qualitative) was performed by in vitro cytokine expression assay using LPS challenge. Statistical analysis were performed using ANOVA (p 0.05) followed by Tukey HSD using SPSS 16.0. Cluster and biplot (PCA) analysis were conducted to classify filters based on generated responses towards all dependent variables simultaneously and to determine its hallmark classifier using PAST software. result on lung flowcytometry analysis as local site, showed that Divine filter DE was triggering immune response involving Gr-1 + , CD8 + and CD4 + T cells, but not for B cells. All filters were equally increased number of lung CD8 + T cells. This result indicating polarized response involving innate and adaptive immunity which refer to cellular mediated immunity. While B and DF filters were tend to trigger immune response involving Gr-1 + , CD8 + T cells and B cells, indicating immune response directed to cellular mediated immunity that followed by humoral immunity via T-helper independent pathway. Based on CD62L differential marker on activation status, DE filter showed highest activation on lung CD4 + and CD8 + T cells and accompanied by an increase in number of CD4 + CD25 + regulatory T cells, indicating balance regulation of activated immune response. Acute exposures of Divine filtered kretek smoke modulated haematological profile shown by significant indication of platelet activation (increased count of PLT and PDW) on DF filter followed by indication of increased platelet production (MPV) on B and DE filters treatment, but this response did not change LED level. Differential count of white blood cells were also increased significantly on all subset population. Significant increase on lymphocyte, monocyte and granulocyte population were marked on B and DF filters treatment, whereas DE filter treatment gave marked increase on lymphocyte and neutrophil population. modulation profile on lung immunocytes (CD4 + and CD8 + T cells, CD4 + CD25 + regulatory T cells and B220 + cells) did not appear on spleen populations, except for spleen Gr-1 + cells on B and DF filters that markedly increased. In general, even if it was not significant, re was a declining trend in number of CD4 + and CD8 + T cells fairly as B220 + cells in spleen. This profile indicating no significant effect on spleen cellular populations as systemic effect of exposures. In vitro cytokine expression study showed that smoke exposures generated from
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/615.952 395 2/ALB/m/041204703 |
| Subjects: | 600 Technology (Applied sciences) > 615 Pharmacology and therapeutics > 615.9 Toxicology |
| Divisions: | S2/S3 > Magister Matematika, Fakultas MIPA |
| Depositing User: | Endro Setyobudi |
| Date Deposited: | 10 Dec 2013 09:01 |
| Last Modified: | 10 Dec 2013 09:01 |
| URI: | http://repository.ub.ac.id/id/eprint/158205 |
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