BKG

Rosmarwati, Ervina (2016) Peran Hsp70 Dan Hmgb1 Dalam Terjadinya Kerusakan Ginjal Pada Model Mencit Malaria Berat. Magister thesis, Universitas Brawijaya.

Indonesian Abstract

Malaria adalah penyakit reemerging, yaitu penyakit infeksi yang sebelumnya sudah terkontrol, kemudian hilang tetapi muncul kembali dengan tampilan yang lebih virulen. Malaria ditemukan terutama di negara-negara tropis dan merupakan salah satu penyebab kematian tertinggi akibat infeksi di Indonesia. Infeksi Malaria berat sering disebabkan oleh Plasmodium falciparum, yang salah satunya bermanifestasi pada organ ginjal, berupa gagal ginjal akut (GGA). Parasit yang menginfeksi eritrosit akan dapat menyebabkan sitoadheren, rosetting, dan sekuestrasi. Mekanisme yang terjadi pada pembuluh darah ini akan menyebabkan iskemia ginjal dan menyebabkan hipoksia. Gangguan iskemik ginjal dapat meningkatkan protein HSP70 dan HMGB1 sebagai respon terhadap iskemia ginjal. HSP70 berfungsi sebagai protein yang membantu proses perbaikan sel, sedangkan HMGB1 berfungsi sebagai sinyal tanda bahaya pada saat terjadi iskemia yang dapat menginduksi inflamasi. Heat Shock Protein 70 (HSP70) bekerja dengan cara memperbaiki protein yang unfolded, mencegah agregasi protein yang terdenaturasi, degradasi protein yang rusak, perbaikan DNA, menghambat apoptosis, stabilisasi cytoskeleton, dan anti inflamasi. HMGB1 berperan sebagai sitokin inflamasi untuk stimulasi maturasi sel dendritik dan bekerja melalui Receptor for Advanced Glycation End Products (RAGE) dan Toll like receptor (TLR), terutama TLR- 2/4. Peran HSP70 dan HMGB1 dalam terjadinya kerusakan ginjal pada model mencit malaria berat masih belum banyak diketahui. Penelitian ini bertujuan untuk mengetahui peran HSP70 dan HMGB1 dalam kerusakan ginjal pada mencit model malaria berat. Penelitian ini merupakan eksperimental laboratorik, post-test only control group menggunakan mencit galur C57BL/6 yang terbagi menjadi kelompok perlakuan dan kelompok kontrol. Kelompok perlakuan terdiri dari 9 ekor mencit yang diinokulasikan Plasmodium berghei 1.106 dalam 0,2 ml darah, sedangkan kontrol negatif adalah mencit yang tidak diinokulasikan Plasmodium berghei. Derajat parasitemia diukur setiap hari dengan cara menghitung jumlah eritrosit yang terinfeksi dalam setiap 1000 eritrosit. Mencit akan dikorbankan pada hari ke-14 untuk diambil ginjalnya. Tingkat kerusakan histopatologi jaringan ginjal dinilai dengan pengecatan hematoksilin eosin (HE), sedangkan tingkat ekspresi HSP70 dan HMGB1 diukur dengan metode imunohistokimia menggunakan antibodi HSP70 dan antibodi HMGB1. Pengamatan dilakukan dibawah mikroskop elektron dengan pembesaran 1000x pada 20 lapangan pandang. Sel yang positif akan mengekspresikan HSP70 dan HMGB1 pada sitoplasma dengan warna coklat. Sel yang dapat mengekspresikan HSP70 dan HMGB1 diantaranya adalah endotel, sel tubulus, sel glomerulus, sel intersisial, dan makrofag. Hasil penelitian dianalisis dengan menggunakan SPSS 22.0 untuk Windows dengan tingkat signifikansi p=0,05 dan taraf kepercayaan 95%. Dari hasil penelitian ini didapatkan perbedaan ekspresi HSP70 yang signifikan antara kelompok perlakuan dan kelompok kontrol (p= 0.000 uji Mann-Whitney). Selain itu, pada kelompok perlakuan dan kelompok kontrol didapatkan perbedaan ekspresi HMGB1 yang signifikan (p= 0.000 uji Mann-Whitney). Hasil uji Korelasi Spearman menunjukkan bahwa terdapat korelasi yang signifikan antara ekspresi HSP70 dan HMGB1 dengan nilai (p= 0.000, R=1.000). Nilai ini menunjukkan hubungan positif antara keduanya, dengan kekuatan hubungan “sangat kuat”. Namun, tidak didapatkan korelasi antara ekspresi HSP70 dengan derajat parasitemia mencit (p = 0.212, R = 0.461) dan tidak ada korelasi antara ekspresi HMGB1 dengan derajat parasitemia mencit (p = 0.212, R = 0.461). Dari hasil penelitian diatas, dapat disimpulkan bahwa HSP70 dan HMGB1 terbukti memiliki peran dalam kerusakan ginjal pada malaria berat. Selain itu, terjadi peningkatan ekspresi HSP70 dan HMGB1 di ginjal pada kelompok perlakuan dibandingkan dengan kelompok kontrol.

English Abstract

Malaria is reemerging diseases, which means an infectious disease that had previously been controlled, then disappear but reappear with more virulent views. Malaria is found mainly in tropical countries and is one of the leading causes of death from infection in Indonesia. Severe malaria infections are often caused by Plasmodium falciparum, which can manifest in the kidneys, such as acute renal Injury (AKI). Parasites that infect erythrocytes can lead to cytoadherence, rosetting and sequestration. These mechanism that occur in blood vessels will cause renal ischemia and hypoxia. Impaired renal ischemia can increase the protein HSP70 and HMGB1 in response to renal ischemia. HSP70 serves as a chaperon and protein that helps cell repair process, while HMGB1 serves as a dangerous signal at the time of ischemia to induce inflammation. Heat Shock Protein 70 (HSP70) works by repairing the unfolded proteins, preventing aggregation of denatured proteins, degrades damaged proteins, DNA repair, inhibits apoptosis, cytoskeleton stabilization, and anti-inflammatory. HMGB1 acts as inflammatory cytokine to stimulate dendritic cell maturation and worked through the Receptor for Advanced Glycation End Products (RAGE) and Toll like receptor (TLR), mainly TLR- 2/4. HSP70 and HMGB1 role in the occurence of kidney damage in severe malaria mice model is still not widely known. This study aimed to determine the role of HSP70 and HMGB1 in kidney damage in severe malaria mice model. This research was an experimental laboratory, post-test only control group using mice strain C57BL/6 which divided into treatment group and control group. Treatment group mice were inoculated with 1.106 of Plasmodium berghei in 0.2 ml of blood. While control group mice were not inoculated with Plasmodium berghei. The degree of parasitaemia was measured every day by counting the number of infected erythrocytes in every 1,000 erythrocytes. Mice have been sacrificed on the 14th day and both kidneys were taken. The level of kidney damage in histopathology was assessed by hematoxylin eosin staining (HE), while the level of HSP70 and HMGB1 expression were measured by immunohistochemistry methods using HSP70 antibody and HMGB1 antibody. Observations were made under the electron microscope with 1000x magnitude on 20 field of view. Positive cells expressed HSP70 and HMGB1 in the cytoplasm with brown color. Cells that expressed HSP70 and HMGB1 include endothelial cells, tubular cells, glomerular cells, interstitial cells, and macrophages. The results were analyzed using SPSS 22.0 for Windows with a significance level of p = 0.05 and 95% confidence level. The results of this study found a significant difference between the expression of HSP70 in treatment group and control group (p= 0.000 Mann-Whitney test). Besides, there was also a significant difference between the expression of HMGB1 in treatment group and control group (p= 0.000 Mann-Whitney test). The Spearman correlation test results showed that there was a significant correlation between the expression of HSP70 and HMGB1 by value (p = 0.000, R = 1.000). This value represented a positive relationship between HSP70 and HMGB1 with the strength of the relationship was "very strong". However, there was no correlation between the expression of HSP70 with parasitaemia in mice (p = 0.212, R = 0.461) and no correlation between the expression of HMGB1 with parasitaemia in mice (p = 0.212, R = 0.461). From the above results, it can be concluded that HSP70 and HMGB1 proved to have role in the occurrence of Acute Kidney Injury in severe malaria. Besides, the expression of HSP70 and HMGB1 in renal treatment group with acute kidney injury (AKI) due to severe malaria proved to be higher than the control group.

Other Language Abstract

UNSPECIFIED

Item Type: Thesis (Magister)
Identification Number: TES/616.936 2/ROS/p/2016/041700799
Uncontrolled Keywords: HEAT SHOCK PROTEINS, KIDNEYS - DIREASES, MALARIA - ANIMAL MODELS
Subjects: 600 Technology (Applied sciences) > 611 Human anatomy, cytology, histology > 611.6 Urogenital systems
Divisions: S2 / S3 > Magister Ilmu Biomedis, Fakultas Kedokteran
Depositing User: Nur Cholis
URI: http://repository.ub.ac.id/id/eprint/1875
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