Studi Potensi Hesperetin sebagai Viral Cellular Entry Inhibitor menggunakan Model Pseudovirus SARS-COV-2

Hayfa Salsabila Harsan, Prof. Dr. apt. Edy Meiyanto, M.Si.; Dr. apt. Endah Puji Septisetyani, M.Sc.

2023 | Skripsi | FARMASI

Agen antivirus dibutuhkan sebagai upaya pengendalian kasus infeksi SARS-CoV- 2 sehingga laju infeksi dapat ditekan. Hesperetin diketahui memilikiaktivitas antivirus yang berpotensi menghambat infeksi virus SARS-CoV-2. Penelitian ini bertujuan mempelajari potensi antivirus dari hesperetin melalui penghambatan viral cellular entry menggunakan model pseudovirus. Hesperetin diuji dengan uji [3-(4,5dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide] (MTT) untuk menentukan konsentrasi non sitotoksik. Hesperetin konsentrasi <100>?M menunjukkan hasil tidak toksik pada model sel 293T dengan persen viabilitas sel >70%. Angiotensin Converting Enzyme 2 (ACE2) dan Transmembrane protease serine2 (TMPRSS2) adalah reseptor yang berperan pada mekanisme masuknya virus SARS-CoV-2. Ekspresi ACE2 dianalisis dengan immunofluorescence staining yang ditandai ekspresi Alexa-594 pada sel 293T. Model sel yang digunakan pada uji entry inhibitor adalah 293T yang ditransfeksi plasmid ACE2 dan TMPRSS2. Pseudovirus SARS-CoV-2 merupakan rekayasa rVSV-?G/G*- GFP-Spike yang inkompeten. Persen sel terinfeksi dilakukan untuk menilai potensi hesperetin menghambat pseudovirus SARS-CoV-2. Nilai persen sel terinfeksi secara berturut-turut pada kontrol, hesperetin 10 ?M, dan hesperetin 100 ?M adalah 43,07%; 18,25%; dan 15,83%. Keseluruhan hasil dianalisis menggunakan One Way Anova Bonferroni (p<0>

Antiviral agents are needed as an effort to control cases of SARS-CoV-2 infection so that the rate of infection can be reduced. Hesperetin is known to have antiviral activity that has the potential to inhibit infection with the SARS-CoV-2 virus. This study aims to study the antiviral potential of hesperetin through inhibition of viral cellular entry using a pseudovirus model. Hesperetin was tested by [3- (4,5dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide] (MTT) test to determine the non-cytotoxic concentration. Hesperetin concentration <100>showed non-toxic results in the 293T cell model with a percent cell viability of >70%. Angiotensin Converting Enzyme 2 (ACE2) and Transmembrane protease serine 2 (TMPRSS2) are receptors that play a role in the entry mechanism of the SARS-CoV-2 virus. ACE2 expression was analyzed by immunofluorescence staining which marked Alexa-594 expression on 293T cells. The cell model used in the entry inhibitor test was 293T transfected with ACE2 and TMPRSS2 plasmids. The SARS-CoV-2 pseudovirus is an incompetent engineered rVSV-?G/G*-GFP- Spike. The percentage of infected cells was carried out to assess the potential of hesperetin to inhibit the SARS-CoV-2 pseudovirus. The percentage value of infected cells respectively in control, 10 ?M hesperetin, and 100 ?M hesperetin was 43.07%; 18.25%; and 15.83%. All results were analyzed using Bonferroni's One Way Anova (p<0>

Kata Kunci : SARS-CoV-2, viral entry inhibitor, antivirus, Hesperetin