PENGARUH ASAM KLOROGENAT (CHLOROGENIC ACID) TERHADAP PERBAIKAN DISFUNGSI MEMORI, EKSPRESI mRNA p53, Bax DAN Bcl-2 PADA TIKUS MODEL DIABETES MELLITUS

ARI SUSILOWATI, Dr. dr. Dwi Cahyani Ratna Sari, M.Kes,PA(K).; dr. Nur Arfian, Ph.D

2021 | Tesis | MAGISTER ILMU BIOMEDIK

Latar Belakang. Diabetes mellitus meningkatkan disfungsi memori. Hiperglikemia dapat menyebabkan stres oksidatif sehingga mengakibatkan apoptosis sinapsis dan neuron di hipokampus. Chlorogenic acid (CGA) dapat menurunkan disfungsi memori, mempengaruhi aktivitas lokomotor, dan memberikan efek anti apoptosis dan anti oksidan. Tujuan Penelitian. Penelitian ini mengkaji pengaruh CGA dalam menurunkan disfungsi memori serta pengaruh CGA dalam ekspresi mRNA p53, Bax, Bcl-2 di hippocampus pada tikus model diabetes. Metode Penelitian. Tiga puluh ekor tikus Rattus norvegicus jantan 150-200 gram dibagi menjadi 6 kelompok Kelompok 1 (Kontrol): tikus dengan pemberian saline (NaCl). Kelompok 2 dan 3 (DM 1,5 dan DM 2) : diinjeksi STZ 60 mg/Kg BB i.p dosis tunggal selama 1,5 dan 2 bulan. Kelompok 4, 5, dan 6: diberi STZ selama 1,5 bulan ditambah CGA. Chlorogenic Acid (CGA) diberikan secara intraperitoneal dengan dosis 12,5 ; 25 dan 50 mg/kg BB selama 14 hari berturut-turut setelah satu setengah bulan di injeksi STZ. Pembelajaran spasial dan kemampuan memori dinilai melalui tes Morris Water Maze. Tingkat ekspresi gen p53, Bax, dan Bcl-2 diperiksa dengan RT-PCR. Apoptosis jaringan diperiksa menggunakan imunohistokimia p53. Analisis data dengan uji parametrik One Way ANOVA dan post hoc LSD. Uji non parametrik kruskal wallis dilanjutkan Mann Whitney-U. Hasil Penelitian. Kelompok DM 2 memiliki waktu tempuh paling lama, yaitu 40,00+-7,70 detik (p=0,021). Kelompok CGA 12,5 memiliki rerata waktu tercepat , yaitu 4,79+- 0,85 detik. Kelompok DM 2 memiliki jarak tempuh terpanjang, yaitu 3,97+-1,07 meter (p=0,011), sedangkan kelompok CGA 12,5 memiliki jarak terpendek, yaitu 0,77+-0,16 meter pada hari ke-5 uji akuisisi Morris Water Maze. Kelompok DM 2 memiliki ekspresi mRNA p53 dan Bax lebih tinggi dibanding semua kelompok CGA. Kelompok Chlorogenic Acid (CGA) 12,5 memiliki ekspresi mRNA Bcl-2 lebih tinggi dari kelompok DM. Kesimpulan. Chlorogenic Acid (CGA) mampu mengurangi disfungsi memori, ditunjukkan dengan waktu latensi dan jarak tempuh yang lebih rendah dari kelompok DM. Chlorogenic Acid (CGA) mampu menurunkan aktivitas apoptosis akibat kondisi diabetes mellitus.

Background. Diabetes mellitus increases memory dysfunction. Hyperglycemia causes oxidative stress resulting in apoptosis of synapses and neurons in the hippocampus. Chlorogenic acid (CGA) can reduce memory dysfunction, affect locomotor activity, and provide anti-apoptotic and anti-oxidant effects. Research purposes. This study examined the effect of CGA in reducing memory dysfunction and the effect of CGA on p53, Bax, and Bcl-2 mRNA expression in the hippocampus in STZ-induced diabetic rats. Research methods. Thirty male Rattus norvegicus rats 150-200 grams were divided into 6 groups. Group 1 (Control): control rats given saline (NaCl). Groups 2 and 3 (DM 1.5 and DM 2): injection of STZ 60 mg/Kg BW i.p single dose for 1.5 and 2 months. Groups 4, 5, and 6: were given STZ for 1.5 months plus CGA. Chlorogenic Acid (CGA) was administered intraperitoneally at a dose of 12.5 ; 25 and 50 mg/kg BW for 14 consecutive days starting one and a half months after STZ injection. Spatial learning and memory skills were assessed through the Morris Water Maze test. The expression levels of p53, Bax, and Bcl-2 genes were examined by reverse transcription polymerase chain reaction (RT-PCR). Tissue apoptosis was examined using p53 immunohistochemistry. Data analysis using One Way ANOVA parametric test and post hoc LSD. Kruskal Wallis non parametric test followed by Mann Whitney-U. Research result. The DM 2 group had the longest travel time, which was 40.00+-7.70 seconds (p=0.021). The CGA 12.5 group had the fastest average time, which was 4.79+- 0.85 seconds. The DM 2 group had the longest distance, which was 3.97+-1.07 meters (p=0.011), while the CGA 12.5 group had the shortest distance, which was 0.77+-0.16 meters on the 5th day of the Morris Water Maze acquisition test. The DM 2 group had p53 and Bax mRNA expression was higher than all CGA groups. The Chlorogenic Acid (CGA) 12.5 group had a higher Bcl-2 mRNA expression than the DM group. Conclusion. Chlorogenic Acid (CGA) was able to reduce memory dysfunction, indicated by lower latency time and mileage than the DM group. Chlorogenic Acid (CGA) is able to reduce apoptotic activity due to diabetes mellitus.

Kata Kunci : Chlorogenic acid (CGA),diabetes mellitus,disfungsi memori, apoptosis, hippocampus, Chlorogenic acid (CGA)