Disertasi

Efek Ekstrak Etanol Biji C. sativum (Coriandrum sativum L) Terhadap Cellular Senescence Pada Organ Kardiovaskular Tikus Induksi Pakan Tinggi Lemak = The effects of ethanolic extract of C. sativum (Coriandrum Sativum L) Seed on Cellular Senescence in the Cardiovascular Organs of High-Fat Diet-Induced Rats.

Latar belakang : Asupan lemak tinggi dapat memicu terjadinya obesitas sebagai faktor risiko penyakit kardiovaskular. Spesies oksigen reaktif berlebihan menyebabkan akumulasi sel senescent. Pengembangan agen senolitik maupun senomorfik merupakan strategi untuk mengatasi faktor risiko dari patologi yang disebabkan oleh cellular senescence melalui jalur anti-apoptosis SCAP (Senescent Cell Anti-apoptotic Pathways) atau faktor SASP (Senescence-Associated Secretory Phenotype). Agen senoterapeutik dari bahan herbal belum banyak dikembangkan, terutama biji C. sativum dengan aktivitas kardioprotektifnya. Oleh karena itu, penelitian ini akan mengevaluasi peran EEBC (Ekstrak Etanol Biji C. Sativum) terhadap sel senescent pada organ kardiovaskular yaitu jaringan jantung, aorta dan lemak di sekitar aorta. Metode : Identifikasi senyawa pada EEBC melalui GC-MS dan LC-MS/MS. Perangkat lunak PyRx, Autodocktools, Visualization Disocvery studio digunakan pada studi in silico pada kelompok SCAP (Bcl-xl, Bcl-2, Bcl-w dan survivin) dan SASP (IL-6, IL-10, IL-1α, TNF-α dan ICAM-1). Pada studi in vivo, dua puluh lima ekor tikus wistar dibagi secara acak menjadi lima kelompok berdasarkan pemberian pakan dan EEBC. Biomarker senescence yaitu aktivitas SA-β-Gal dan lipofuscin diperiksa pada organ kardiovaskular jantung, aorta dan lemak di sekitar aorta. Jaringan yang menunjukan marker senescence dievaluasi dengan pengukuran inisiator senescence (ELISA p21), anti-apoptosis (ELISA Bcl-xl) beserta apoptosis (Uji TUNEL), histologi lipid droplet dan morfologi brown adipocyte. Pengukuran SASP melalui uji LUMINEX. Hasil : EEBC tidak mempengaruhi aktivitas SA-β-Gal dan lipofuscin pada jaringan jantung dan aorta. EEBC memberikan pengaruh terhadap penurunan aktivitas SA- β-Gal dan lipofuscin pada jaringan lemak di sekitar aorta. EEBC menurunkan kadar p21 baik pada jaringan jantung maupun lemak di sekitar aorta. EEBC juga menurunkan kadar Bcl-xl, sehingga cenderung meningkatkan apoptosis pada jaringan lemak sekitar aorta. Pemeriksaan histologi menunjukan terjadinya penurunan area lipid droplet dan peningkatan jumlah sel lemak dengan morfologi brown adipocyte pada jaringan lemak di sekitar aorta. EEBC tidak mempengaruhi faktor SASP. Kesimpulan : EEBC dapat mencegah early senescence pada jaringan lemak sekitar aorta, tetapi tidak pada jaringan jantung dan aorta.
Kata kunci : C. sativum, pakan tinggi lemak, early senescence, senoterapeutik.


Background : High fat intake can lead to obesity as a risk factor for cardiovascular disease. Excessive reactive oxygen species cause the accumulation of senescent cells. The development of senolytic and senomorphic agent is a strategy to address risk factors of pathology linked to cellular senescence through the anti-apoptotic pathway (SCAP) or SASP factor. Senotherapeutic agent from herbal have not been generally developed, particularly C. sativum seeds with cardioprotective properties. Therefore, the study intended to evaluate the role of EECS (Ethanolic Extract of Coriandrum sativum L. ) on senescent cells in cardiovascular organs such as the heart, aorta and adipose tissue surrounding the aorta using in silico and in vivo methods. Method: Identification of compounds in EECS through GC-MS and LC-MS. PyRx, Autodocktools, Visualization Discovery studio software were used in the in silico study on the SCAP group (Bcl-xl, Bcl-2, Bcl-w and survivin) and SASP (IL-6, IL- 10, IL-1α, TNF-α and ICAM-1) group. In vivo study, twenty-five wistar mice were randomly divided into 5 groups based on administration of diet and EECS. Senescence biomarkers such as SA-β-Gal and lipofuscin activity were examined in the cardiovascular organs of the heart, aorta and adipose tissue surrounding the aorta. Tissues showing senescence markers were evaluated by measuring senescence inducer (p21 ELISA), anti-apoptosis (Bcl-xl ELISA), apoptosis (TUNEL assay), lipid droplet and brown adipocyte morphology (Histology). Measurement of SASP through Luminex assay. Results: EECS has no effect on the activity of SA-β-Gal and lipofuscin in the heart and aortic tissue. EECS reduces SA-β-Gal and lipofuscin activity in adipose tissue surrounding the aorta. EECS reduced levels of p21 in the heart and adipose tissue around aorta. EECS also decreases Bcl-xl levels which tends to increase apoptosis in adipose tissue. Histological examination showed a decrease in lipid droplet area and an increase in brown adipocyte-morphology cell numbers in adipose tissue around aorta. EECS does not affect SASP factors. Conclusion: EECS can prevent early senescence on the adipose tissue around the aorta, but not in the heart and aortic tissues.
Keywords : C. sativum, high-fat diet, early senescence, senotherapeutic.