Alifia, Lustyafa Inassani and Prof. Dr. dr. Loeki Enggar Fitri, M. Kes, Sp.ParK and Prof. Agustina Tri Endharti, S.Si, PhD (2022) Pengaruh Pemberian Probiotik Lactobacillus casei dan Bifidobacterium longum Terhadap Derajat Parasitemia, Ekspresi CD103, dan FoxP3 Intestinal Mencit C57BL/6 yang Diinfeksi Plasmodium berghei. Magister thesis, Universitas Brawijaya.
Abstract
Malaria yang disebabkan oleh parasit darah genus Plasmodium masih menjadi masalah kesehatan global. Beberapa studi telah mengungkapkan bahwa timbulnya beberapa penyakit, termasuk penyakit infeksi, berhubungan dua arah dengan perubahan lingkungan usus, terutama perubahan komposisi mikrobiota usus (gut microbiota), yang berasosiasi dengan disbiosis usus. Selama fase intraeritrositik pada siklus Plasmodium, eritrosit terinfeksi mampu bersirkulasi berbagai jaringan melalui sirkulasi darah dan menimbulkan kaskade respons imun, termasuk sistem kekebalan intestinal. Kondisi disbiosis akibat infeksi Plasmodium dapat mempengaruhi respon imun usus, produksi sitokin abnormal, dan keparahan penyakit malaria. Beberapa sel imun yang berperan dalam pembentukan imunitas usus adalah sel dendritik (DC) usus dan sel T regulator (Treg) usus. Ketika probiotik mencapai usus, fragmen probiotik diinternalisasi oleh sel microfold, melintasi epitel dan ditangkap oleh makrofag dan DC usus, yang selanjutnya akan melepaskan sitokin . DC usus akan mencapai Mesenteric Lymph Node (MLN) dan kemudian membantu dalam diferensiasi sel Treg. DC usus memiliki penanda permukaan CD103 sedangkan pada sel Treg usus mengekspresikan FoxP3 yang merupakan faktor transkripsi dari sel Treg. Tujuan dari penelitian ini adalah untuk membuktikan pengaruh pemberian probiotik Lactobacillus casei (L. casei) dan Bifidobacterium longum (B. longum) terhadap derajat parasitemia, ekspresi CD103 dan FoxP3 intestinal mencit C57BL/6 yang diinfeksi Plasmodium berghei (P. berghei). Metode penelitian ini menggunakan desain true experimental dengan pendekatan post-test only control group design. Subyek mencit secara acak dibagi menjadi kelompok yang diberi PBS atau kontrol negatif (K-), kelompok yang diinjeksi P. berghei atau kelompok kontrol positif (K+), dan kelompok yang diberi intervensi probiotik L. casei dan B. longum P1 baik tunggal maupun kombinasi yaitu diberi P. berghei + L. casei (KP1), diberi P. berghei + B. longum (KP2), dan yang diberi P. berghei + L .casei + B.longum (KP3). L. casei dan B. longum diberikan melalui galvage dari H-5 sampai hari ke-6 pasca-infeksi P. berghei. Inokulasi P. berghei dilakukan pada kelompok perlakuan secara intraperitoneal dengan dosis 5x106. Parameter yang diamati adalah derajat parasitemia, ekspresi CD103, dan FoxP3 intestinal mencit yang menggunakan immunofluoresens. Semua kelompok perlakuan mengalami peningkatan derajat parasitemia dari hari ke-2 sampai hari ke-6 pasca-infeksi. Kelompok yang mendapat intervensi probiotik B. longum saja menunjukkan derajat parasitemia paling rendah sampai hari ke-6 dibandingkan kelompok perlakuan lainnya. Uji Kruskal Wallis pada hari ke-2 inokulasi P. berghei menunjukkan perbedaan signifikan antara kelompok perlakuan (p=0,001). viii Ekspresi CD103 meningkat pada kelompok KP1, KP2, dan KP3 dibandingkan kelompok kontrol positif (p=0,01). Kelompok KP2 dan KP3 menunjukkan peningkatan ekspresi FoxP3 dibandingkan dengan kontrol positif (p=0,002). Pemberian probiotik B. longum berpotensi menghambat pertumbuhan parasitemia melalui respon imun lokal pada usus, namun kolonisasinya di usus perlu diteliti lebih dalam untuk mengetahui manfaatnya pada tubuh inang. Perawatan probiotik dapat mempengaruhi tingkat parasitemia dan kekebalan usus selama infeksi malaria
English Abstract
Malaria caused by blood parasites of the genus Plasmodium is still a global health problem. Several studies have revealed that the emergence of several diseases, including infectious diseases, is related in two ways to changes in the intestinal environment, especially changes in the composition of the gut microbiota, which are associated with intestinal dysbiosis. During the intraerythrocytic phase of the Plasmodium cycle, infected erythrocytes are able to circulate to various tissues through the bloodstream and elicit a cascade of immune responses, including the intestinal immune system. The condition of dysbiosis due to Plasmodium infection can affect the intestinal immune response, abnormal cytokine production, and the severity of malaria.The purpose of this study was to prove the effect of probiotics Lactobacillus casei (L. casei) and Bifidobacterium longum (B. longum) on the degree of parasitemia, expression of CD103 and intestinal FoxP3 in C57BL/6 mice infected with Plasmodium berghei (P. berghei). Several immune cells that play a role in the formation of intestinal immunity are intestinal dendritic cells (DCs) and intestinal regulatory T cells (Treg). When probiotics reach the gut, the probiotic fragments are internalized by microfold cells, cross the epithelium and are captured by intestinal macrophages and DCs, which in turn release cytokines. Intestinal DCs will reach the Mesenteric Lymph Node (MLN) and then assist in Treg cell differentiation. Intestinal DCs have the surface marker CD103 whereas intestinal T cells express FoxP3 which is a transcription factor of T cells. The purpose of this study was to prove the effect of L. casei and B. longum on the degree of parasitemia, expression of CD103 and intestinal FoxP3 in C57BL/6 mice infected with Plasmodium berghei. This research method uses a true experimental design with a post-test only control group design approach. Mice subjects were randomly divided into five group, which the first group given PBS alone or a negative control (K-), second group injected with P. berghei only or a positive control group (K+), and group was given both L. casei and B. longum probiotic intervention either single. or a combination of P. berghei + L. casei (KP1), P. berghei + B. longum (KP2), and those given P. berghei + L.casei + B.longum (KP3). L. casei and B. longum were administered via galvage from D-5 to 6th day post-infection with P. berghei. Inoculation of P. berghei was carried out in the treatment group intraperitoneally at a dose of 5x106. Parameters observed were the x degree of parasitemia, expression of CD103, and intestinal FoxP3 of mice using immunofluorescence. All treatment groups experienced an increase in the degree of parasitemia from day 2 to day 6 post-infection. The group that received the B. longum probiotic intervention alone showed the lowest degree of parasitemia until the 6th day compared to the other treatment groups. The Kruskal Wallis test on day 2 of P. berghei inoculation showed a significant difference between the treatment groups (p=0.001). CD103 expression increased in the KP1, KP2, and KP3 groups compared to the positive control group (p=0.01). The KP2 and KP3 groups showed increased expression of FoxP3 compared to the positive control (p=0.002). Giving B. longum probiotics has the potential to inhibit the growth of parasitemia through a local immune response in the intestine, but its colonization in the intestine needs to be studied more deeply to determine its benefits to the host body. Probiotic treatment may affect parasitemia levels and intestinal immunity during malarial infection.
Other obstract
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| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | 0422060193 |
| Subjects: | 600 Technology (Applied sciences) > 616 Diseases > 616.02 Special topics of disease > 616.025 Medical emergencies / Emergency medicine / Emergency nursing / Triage (Medicine) |
| Divisions: | S2/S3 > Magister Ilmu Biomedis, Fakultas Kedokteran |
| Depositing User: | Endang Susworini |
| Date Deposited: | 13 Jul 2023 07:21 |
| Last Modified: | 13 Jul 2023 07:21 |
| URI: | http://repository.ub.ac.id/id/eprint/201835 |
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