Pengaruh Pemberian Exosome (HUC-MSC-Exo) terhadap Gambaran Cedera Tubulus, Kadar BUN, dan Apoptosis Sel Epitel Ginjal pada Tikus Model Cedera Iskemia Reperfusi (IR) Periode Kronis

Ghea Farmaning Thias Putri, dr. Nur Arfian, Ph.D ; dr. Ratih Yuniartha, Ph.D

2024 | Tesis | S2 Ilmu Kedokteran Dasar dan Biomedis

Latar Belakang: Chronic Kidney Disease (CKD) merupakan permasalahan kesehatan yang serius di dunia. CKD seringkali berasal dari penyakit ginjal akut (AKI), yang umumnya ditandai oleh cedera iskemia reperfusi (IR), yang dapat memicu proses apoptosis. Terapi konvensional untuk CKD belum mampu secara efektif memperbaiki kerusakan ginjal yang signifikan, sehingga meningkatkan kebutuhan akan pendekatan baru. Human Umbilical cord MSC-derived Exosome (HUC-MSC-Exo) menunjukkan potensi sebagai terapi inovatif yang menjanjikan.

Tujuan: Penelitian ini bertujuan untuk mengetahui pengaruh pemberian Exosome dari Human Umbilical cord MSC-derived Exosome (HUC-MSC-Exo) terhadap gambaran cedera tubulus, kadar BUN, dan apoptosis sel epitel ginjal pada tikus model cedera iskemia reperfusi (IR) periode kronis.

Metode: Penelitian ini melibatkan subjek penelitian berupa hewan percobaan yang dimanipulasi dan dibandingkan dengan kelompok kontrol. Dalam penelitian ini, digunakan tikus galur Wistar Rattus norvegicus sebanyak 25 ekor, dengan jenis kelamin jantan, usia 2-3 bulan, dan berat 200-250 gram. Subyek penelitian dibagi menjadi 5 kelompok: Kelompok SO, kelompok IR, kelompok IR+ Exo1 (24,15 ?g), kelompok IR+Exo2 (48,30 ?g), dan kelompok IR+Exo3 (96,61 ?g). Evaluasi cedera tubulus dilakukan dengan pewarnaan PAS, pemeriksaan BUN, penilaian ekspresi mRNA Bax, Bcl-2, rasio mRNA Bax/Bcl-2, serta analisis imunohistokimia Bax.

Hasil: Pada tikus model cedera iskemia reperfusi (IR) kronis yang diberikan exosome, cedera tubulus ginjal tampak lebih baik dan sitoplasma sel tubulus ginjal lebih dominan berwarna cokelat dibandingkan kelompok tanpa exosome. Kadar BUN dan ekspresi mRNA Bax tidak menunjukkan perbedaan signifikan. Ekspresi mRNA Bcl-2 lebih rendah pada kelompok Exo-2 dan Exo-3, dan rasio ekspresi mRNA Bax/Bcl-2 lebih rendah dibandingkan tikus tanpa exosome.

Kesimpulan: Pemberian exosome pada ginjal tikus model cedera iskemia reperfusi kronis memperbaiki cedera tubulus dan dominasi warna cokelat pada sitoplasma, dengan ekspresi mRNA Bcl-2 dan rasio Bax/Bcl-2 lebih rendah, tanpa perbedaan signifikan pada kadar BUN dan ekspresi mRNA Bax.

Background: Chronic Kidney Disease (CKD) is a severe health problem worldwide. This disease often originates from Acute Kidney Injury (AKI), which is generally characterized by Ischemia-Ieperfusion (IR) injury, which can trigger apoptosis. Conventional therapies for CKD have not been able to effectively repair significant kidney damage, increasing the need for new approaches. Human Umbilical Cord MSC-derived Exosome (HUC-MSC-Exo) shows potential as a promising innovative therapy.

Objective: This research aims to investigate the effects of administering Exosomes derived from Human Umbilical Cord Mesenchymal Stem Cells (HUC-MSC-Exo) on tubular injury, BUN levels, and epithelial cell apoptosis in a rat model of chronic ischemia-reperfusion (IR) injury.

Method: This research involved research subjects in the form of experimental animals who were manipulated and compared with a control group. In this study, 25 Wistar rats (Rattus norvegicus), male, 2-3 months old, and weighing 200-250 grams were used. The subjects were divided into five groups: SO group, IR group, IR+ Exo1 group (24.15 ?g), IR+Exo2 group (48.30 ?g), and IR+Exo3 group (96.61 ?g). Evaluation of tubular injury was carried out by Periodic Acid-Schiff (PAS) staining, BUN examination, assessment of Bax, Bcl-2, ratio Bax/Bcl-2 mRNA expression, and Bax immunohistochemical analysis.

Results: In a chronic ischemia-reperfusion (IR) injury model in rats, those treated with exosomes exhibited improved tubular injury and a greater presence of brown coloration in the cytoplasm of kidney tubular cells compared to the group without exosome treatment. There were no significant differences in BUN levels and mRNA expression of Bax. The mRNA expression of Bcl-2 was lower in the Exo-2 and Exo-3 groups, and the Bax/Bcl-2 mRNA expression ratio was lower compared to rats not treated with exosomes.

Conclusion: Administration of exosomes in a chronic ischemia-reperfusion injury model in rats improved tubular injury and increased the presence of brown coloration in the cytoplasm. This treatment resulted in lower mRNA expression of Bcl-2 and a lower Bax/Bcl-2 mRNA expression ratio, with no significant differences in BUN levels and mRNA expression of Bax.

Kata Kunci : Iskemia reperfusi, exosome, apoptosis, Bax, Bcl-2.