THE ROLE OF CHLOROGENIC ACID IN HEPATIC FIBROSIS INDUCED BY CARBON TETRACHLORIDE IN MICE MODEL: EXPLORATION OF MACROPHAGE NUMBER, MONOCYTES CHEMOATTRACTANT PROTEIN-1 EXPRESSION (MCP-1), AND ALBUMIN LEVEL

Mohammad Imad Awad Ghannam, dr. Nur Arfian, PhD.; Dr.dr. Dwio Cahyani Rat Sari, MKes, SpPA(K).

2017 | Tesis | S2 Ilmu Kedokteran Dasar dan Biomedis

Latar belakang: Fibrosis hepar merupakan jaringan parut yang disebabkan akibat cedera kronis. Hingga saat ini masih belum ditemukan terapi yang efektif untuk fibrosis hepar. Asam klorogenat (chlorogenic acid, CGA) merupakan senyawa fenolik yang memiliki aktivitas antiinflamasi dan antioksidan. Tujuan: Mengkaji perubahan ekspresi Monocyte Chemoattractant Protein-1 (MCP-1), jumlah makrofag, dan kadar serum albumin pada mencit dengan fibrosis hepar setelah pemberian asam klorogenat. Metode: Penelitian ini menggunakan mencit jantan galur Swiss (30 gram, 3 bulan) sebanyak 30 ekor. Mencit dibagi menjadi 6 kelompok yaitu, kontrol, CCl4 dilakukan dengan cara melarutkan CCl4(Merck MAB13426) 0.5 mg/kg BB dalam minyak zaitun (1:1), CGA (Sigma-Aldrich, C3878-1G) dengan dosis 42 mg/kgBB, CGA 63 mg/kgBB, and CGA 84 mg/kgBB. Mencit diterminasi pada minggu ke-4. Selanjutnya dilakukan pemeriksaan jumlah makrofag dilakukan dengan metode IHC, pemeriksaan MCP-1 dilakukan dengan reverse transcription PCR (RT-PCR). Hasil: Pemberian CGA dapat menurunkan jumlah makrofag (p<0.05) dan mRNA MCP-1 pada mencit yang diinduksi fibrosis hepar menggunakan CCl4, dan meningkatkan kadar serum albumin (p<0.05) melalui efek antioksidan dan antiinflamasi. Kesimpulan: Pemberian asam klorogenat dapat menurnkan jumlah makrofag dan MCP-1 dan meningkatkan kadar serum albumin pada mencit dengan fibrosis hepar.

Background: Liver fibrosis is a scaring process related to chronic liver injury of all causes and as yet no truly effective treatment is available. Chlorogenic acid (CGA) is a phenolic compound and exerts anti-inflammatory and anti-oxidant activities. This study will examine the effects of chlorogenic acid (CGA) on liver fibrosis induced by carbon tetrachloride (CCl4) and explored the possible mechanisms of action. Experimental animals were used in this study male mice, Swiss animal strain weighing30+-20 g, the age of mice is 3 months, total number of samples is 30, the main material used in this study is CCl4 (Merck MAB13426) 0.5 mg/kg BW dissolved in olive oil (1: 1) and CGA (Sigma-Aldrich, C3878-1G), 42 mg/kg BW, 63 mg/kg BW, and 84 mg/kg BW. Mice were divided into 6 groups. CCl4 dose 0.5 mg/kg given dissolved along with the olive oil in the ratio 1:1, twice per week, for 4 weeks of treatment injection intraperitoneally, while the CGA doses are 42 mg/kg, 63 mg/kg and 84 mg/kg given orally daily for 4 weeks. This study examined the effects of chlorogenic acid (CGA) on liver fibrosis induced by carbon tetrachloride (CCl4) and explored the possible mechanisms of action through the exploration the effect of CGA on macrophage number, monocyte chemoattractant protein 1 expression (MCP-1), and albumin level. The results showed the CGA could reduce the effect of CCl4 in liver fibrosis in mice model. Objective: to investigate the effect of chlorogenic acid on the macrophage number, chemoattractant monocyte protein-1 expression, and on the albumin level in liver fibrosis caused by carbon tetrachloride. Method: Mice were treated in Laboratory of Anatomy Faculty of Medicine, Universitas Gadjah Mada, for 4 weeks, terminated after 24 hours of treatment ends, terminations carried out by the method of cervical dislocation, were anesthetized previously with ketamine (a dose of 75 mg/kg i.m.), then performed dissection in the abdominal region at midline and perfused the liver organ was taken with formalin 10%. Then performed immunohistochemistry staining, reverse transcription PCR (RT-PCR) to examine MCP-1 expression pathway, and measured the albumin level. Result: the results of this study showed that the administration of CGA could reduce significantly the macrophage number in liver fibrosis induced by CCl4 and increased significantly the serum of albumin levels through improving the antioxidant and anti-inflammatory activities in liver fibrosis induced by CCl4, however the result of MCP-1 expression showed there was no significant effect by CGA on MCP-1 expression in liver fibrosis induced by CCl4. Conclusion: Group with CGA treatment had lower macrophage number compared to CCl4 in liver fibrosis induced by CCl4.The MCP-1 mRNA expression was not significant after the CGA administration in liver fibrosis induced by CCl4. The administration of CGA increase significantly the serum of albumin level in liver fibrosis induced by CCl4.

Kata Kunci : liver fibrosis, Chlorogenic Acid, Carbon tetrachloride, Oxidative Stress, Chemokines.