Pengaruh Pemberian Human Umbilical Cord Mesenchymal Stem Cells Exosomes Terhadap Ekspresi mRNA eNOS, ppET-1, dan Fibrosis Perivaskular pada Ginjal Tikus Model Cedera Iskemia Reperfusi Fase Kronis

Nora Eny Zulikha, dr. Nur Arfian, PhD; dr. Ratih Yuniartha, PhD

2024 | Tesis | S2 Ilmu Kedokteran Dasar dan Biomedis

Latar Belakang: Peningkatan reakctive oxygen species (ROS) pada ischemia reperfusion injury menyebabkan terjadinya stress oksidatif dan inflmasi kronis yang berujung pada cedera vaskular sehingga terjadi disfungsi endotel yang berujung pada fibrosis. Human umbilical cord mesenchymal stem cells derived exosomes memiliki efek dalam perbaikan cedera jaringan.
Tujuan: Untuk mengkaji pengaruh pemberian HUCMSC- Exo terhadap ekspresi mRNA ppET-1 dan eNOS serta fibrosis perivaskular pada ginjal tikus model cedera iskemia reperfusi fase kronis
Metode: Penelitian ini menggunakan desain penelitian posttest-only control group design. Terdiri dari 5 kelompok perlakuan, kelompok SO, kelompok IRI, kelompok IRI + HUCMSC-exo total protein 24,15 µg (Exo-1), kelompok IRI + HUCMSC- exo total protein 48,30 µg (Exo-2) dan kelompok IRI + HUCMSC-exo total protein 96,61 µg (Exo-3). Masing-masing kelompok terdiri dari 5 ekor tikus putih jantan (Rattus norvegicus) galur Wistar. Gambaran histologi ginjal diamati pada preparat dengan pewarnaan Sirius Red, perbesaran 400x. Ekspresi mRNA eNOS dan ppET-1 diuji menggunakan RT-PCR. Analisis statistik menggunakan software SPSS, uji normalitas Shapiro-Wilk, dan uji Anova sebagai uji hipotesis.
Hasil: Ekspresi mRNA eNOS lebih tinggi pada kelompok Exo-3 berbeda signifikan dibandingkan dengan kelompok IRI (p=0.002). Ekspresi mRNA ppET-1 lebih rendah pada kelompok Exo-1 dan Exo-3 berbeda signifikan dibandingkan dengan kelompok IRI (p=0,038; p=0.004). Fraksi area fibrosis perivaskular lebih sedikit pada kelompok Exo-3 berbeda signifikan dibandingkan dengan kelompok IRI (p=0.000). IRI menyebabkan tingginnya fibrosis perivascular dan ekspresi mRNA ppET-1 serta rendahnya ekspresi mRNA eNOS.
Kesimpulan: The administration of HUCMSC-Exo to the IRI rats group result in increased eNOS mRNA expression, decreased ppET-1 mRNA expression and decreased perivascular fibrosis area fraction..
Kata kunci: eNOS, ppET-1, IRI, fibrosis perivaskular, ginjal.

Background: Increased reactive oxygen species (ROS) in ischemia reperfusion injury causes oxidative stress and chronic inflammation which ends in vascular injury resulting in endothelial dysfunction which ends in fibrosis. Exosomes derived from human umbilical cord mesenchymal stem cells have effects in repairing tissue injury
Objective: To examine the effect of HUCMSC-Exo administration on ppET-1 and eNOS mRNA expression as well as perivascular fibrosis in rat kidneys in the chronic phase ischemia reperfusion injury model.
Method: This study used a posttest-only control group design. It consisted of 5 treatment groups, SO group, IRI group, IRI + HUCMSC-exo total protein 24.15 ?g (Exo-1), IRI + HUCMSC-exo total protein 48.30 ?g (Exo-2) group and IRI group + HUCMSC-exo total protein 96.61 (Exo-3). Each group consisted of 5 male white rats (Rattus norvegicus) Wistar strain. The histology of the kidney was observed in preparations with Sirius Red staining, 400x magnification. eNOS and ppET-1 mRNA expressions were tested using RT-PCR. The statistical analysis utilized SPSS software, Shapiro-Wilk normality test, and Anova test as the hypothesis testing.
Results: eNOS mRNA expression higher in the Exo-3 group which was significantly different compared to the IRI group (p=0.002). ppET-1 mRNA expression lower in the Exo-1 and Exo-3 groups significantly different compared to the IRI group (p=0,038; p=0.004). The area fraction of perivascular fibrosis in the Exo-3 group was slightly different compared to the IRI group (p=0.00). IRI causes high perivascular fibrosis and ppET-1 mRNA expression and low eNOS mRNA expression.
Conclusion: The administration of HUCMSC-Exo to the IRI rats group result in increased eNOS mRNA expression, decreased ppET-1 mRNA expression and decreased perivascular fibrosis area fraction
Key words: eNOS, ppET-1, IRI, perivascular fibrosis, kidney.

Kata Kunci : eNOS, ppET-1, IRI, perivascular fibrosis, kidney.