Pengaruh Vitamin C Terhadap Viabilitas Sel Endotel dan Disfungsi Endotel Pada Kultur HUVECs yang Dipapar CdCl2 Melalui NO dan MDA.

Sofiana, Kristianningrum Dian and Prof. dr. M. Aris Widodo,, MS, Sp. FK, Ph.D, and Dr. Husnul Khotimah,, S.si, M.Kes. (2018) Pengaruh Vitamin C Terhadap Viabilitas Sel Endotel dan Disfungsi Endotel Pada Kultur HUVECs yang Dipapar CdCl2 Melalui NO dan MDA. Magister thesis, Universitas Brawijaya.

Abstract

Penyebab kematian tertinggi di dunia disebabkan oleh penyakit kardiovasuler yang diawali dengan terjadinya disfungsi endotel.Beberapa studi mengungkapkan ada kaitan antara logam berat kadmium dengan kelainan vaskular. Kadmium logam berat yang mudah mengkontaminasi lingkungan dengan sumber paparan melalui bahan bakar minyak, kosmetik, pupuk, produksi baja,rokok dan inecerator. Kadmium memasuki tubuh melalui inhalasi dari asap rokok ,udara yang makanan dan minuman yang tercemar kadmium. Beberapa efek kadmium terhadap kesehatan antara lain menyebabkan kanker, penyakit ginjal,penyakit saluran nafas, tulang dan otot . Penelitian terbaru secara epidemiologi menemukan kaitan antara kadar kdmium dalam serum darah maupun urin dengan kejadian penyakit vaskular. Kadmium menyebabkan stres oksidatif dengan beberapa mekanisme antara lain melalui reaksi fenton,menghambat kompleks mitokondria, dan menurunkan antioksidan intraselular. Stress Oksidatif meningkatan ROS (Reaktif Oksigen Species) ,radikal Hydroxil yang bereaksi dengan lipid tak jenuh akan menghasilkan peroksidasi lipid dengan hasil sekunder malondialdehid (MDA). Superoxide bereaksi dengan NO (Nitrit Oxide) membentuk peroxynitrit yang akan menurunkan kadar NO. Radikal Oksigen juga mempengaruhi transkripsi dari DNA yang mengakibatkan penurunan viabilitas sel. Vitamin C bersifat antioksidan yang bersifat larut air, mudah ditemukan pada berbagai jenis buah buahan dan memiliki fungsi enzimatik maupun non enzimatik. Beberapa studi menunjukan Vitamin C berperan dalam fungsi vaskular diantaranya memodulasi vasorelaksasi dengan meningkatakan sintesis atau bioavailabilitas NO melalui ascorbate mediated denitrosylation dan fosforilasi eNOS. Peranan lainnya mengatur transkripsi yang mengkode protein termasuk angiogenesis dan proliferasi sel. Sebagai antioksidan vitamin ini memiliki kemampuan mereduksi atau radikal scavenger pada beberapa radikal bebas dan ROS yang patologis. Gugus enadiol yang terletak pada atom C2 dan C3 pada Vitamin C menyebabkan mampu menangkap radikal hydroksil. Sedangkan untuk radikal bebas oksigen ditangkap dengan atau tanpa bantuan enzim katalisator. Penelitian ini bertujuan untuk melihat pengaruh dari vitamin C pada viabilitas sel endotel dan Disfungsi Endotel melalui kadar NO dan MDA HUVECs ( Human Umbilical Vein Endhotel Cells) yang diinduksi CdCl2 Penelitian ini menggunakan HUVECs konfluen 70% yang dibagi dalam lima kelompok perlakuan yaitu: kontrol, paparan CdCl2, dan kelompok paparan CdCl2 (24,154 μg/L) yang ditambahkan Vitamin C 50,100, 200 μM secara berurutan. Viabilitas sel dianalisis menggunakan MTT assay, Kadar NO diukur dengan Greiss reagen, sedangkan MDA sel diukur dengan menggunakan metode TBARs. Hasil penelitian menunjukan bahwa viabilitas sel endotel mengalami peningkatan dengan penambahan Vitamin C konsentrasi 100 dan 200 μM. Penambahan Vitamin C cenderung meningkatkan kadar NO dibanding dengan kelompok CdCl2 meskipun tidak berbeda signifikan. Kadar MDA HUVECs mengalami penurunan pada pemberian Vitamin C konsentrasi 50 dan 100 μM, sedangkan konsentrasi 200 μM meningkatkan kadar MDA.Dapat Disimpulkan bahwa Vitamin C konsentrasi 100 μM mampu meningkatkan viabilitas sel, kadar No dan menurunkan kadar MDA.

English Abstract

The highest cause of death in the world is caused by cardiovascular disease that begins with the occurrence of endothelial dysfunction. Several studies revealed that there is a link between heavy metal such as cadmium with vascular abnormalities. Cadmium is a heavy metal substance that easily contaminates the environment through fuel oil, cosmetics, fertilizers, steel production, cigarettes and inecerators as its exposure source. Cadmium enters the body through inhalation of smoke from cigarette, air or food and drink that contaminate with cadmium. Some cadmium effects on health include cancer, kidney disease, and respiratory diseases, bones and muscles disease. Recent epidemiological studies have found a link between cadmium levels in both serum and urine with the incidence of vascular disease. Cadmium causes oxidative stress with several mechanisms, such as through fenton reactions, inhibiting mitochondrial complexes, and decreasing intracellular antioxidants. Oxidative Stress increases ROS (Reactive Oxygen Species). Hydroxyl radical is one of the examples of ROS that if it reacts with unsaturated lipids will produce lipid peroxidation with malondialdehid (MDA) as the secondary results. Superoxide that reacts with NO (Nitric Oxide) will form peroxynitrite that might lower the NO levels. Oxygen radical also affects the transcription of DNA and resulting in decreased cell viability. Vitamin C is a water-soluble antioxidant, a vitamin that's easy to find in various types of fruits and has both enzymatic and non enzymatic functions. Several studies have shown that Vitamin C plays a role in vascular function including modifying vasorelaxation by increasing NO synthesis or NO's bioavailability through ascorbate mediated denitrosylation and eNOS phosphorylation. Another role of Vitamin C is to regulate the transcription that encodes proteins including angiogenesis and cell proliferation. As an antioxidant, this vitamin has the ability to reduce or become a radical scavenger in some free radicals and pathological ROS. The enadiol group located on C2 and C3 at Vitamin C lead the vitamin to have the ability to capture hydroxyl radicals. Meanwhile, free radicals of oxygen are captured with or without the support from catalytic enzymes. This research is meant to find out the effect of vitamin C on endothelial cell viability and endothelial dysfunction through NO levels and MDA in HUVECs (Human Umbilical Vein Endhotel Cells) induced by CdCl2. This research used 70% of confluent HUVECs divided into five treatment groups: controll group, CdCl2-exposed group, and CdCl2-exposed group (24,154 μg/L) added with Vitamin C 50,100, and 200 μM respectively. Cell viability was analyzed using an MTT assay, NO content was measured with Greiss reagents, whereas MDA cells were measured using the TBARs method. The research results showed that endothelial cell viability increased with the addition of Vitamin C concentrations of 100 and 200 μM. The addition of Vitamin C tended to increase NO levels compared to the CdCl2 group although it didn't show significant difference. The MDA levels of HUVECs were decreased in the allotment of Vitamin C in 50 and 100 μM concentrations. Meanwhile, 200 μM of Vitamin C concentrations of increased the MDA levels. It can be concluded that 100 μM concentration of Vitamin C is be able to increase the cell viability and NO levels, and decrease the MDA levels.

Item Type:	Thesis (Magister)
Identification Number:	TES/615.328/SOF/p/2018/041802332
Uncontrolled Keywords:	HUVECs, Cadmium, Vitamin C, Viabilitas sel, NO, MDA,HUVECs, Cadmium, Vitamin C, Cell Viability, NO, MDA
Subjects:	600 Technology (Applied sciences) > 615 Pharmacology and therapeutics > 615.3 Organics drugs > 615.32 Drugs derived from plants and mikroorganisms > 615.328 Vitamins
Divisions:	S2/S3 > Magister Ilmu Biomedis, Fakultas Kedokteran
Depositing User:	soegeng sugeng
Date Deposited:	05 Aug 2022 07:42
Last Modified:	05 Aug 2022 07:42
URI:	http://repository.ub.ac.id/id/eprint/193015

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