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The Effect of Lipopolysaccharide Challenge in RAW 264.7 Cells on Nitric Oxide Release and Cell Viability Suprapto, Ratih Paramita; Kusumastuty, Inggita; Rizal, Ardian; Adi Nugroho, Dwi
Jurnal Kedokteran Brawijaya Vol. 33 No. 2 (2024)
Publisher : Fakultas Kedokteran Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jkb.2024.033.02.2

Abstract

Lipopolysaccharide (LPS) is a major component of a gram-negative bacterial wall that is widely used and well-established to induce inflammation in vitro. In addition, the in vitro model using RAW 264.7 cells is the most commonly applied in screening the anti-inflammatory and elucidating the pathophysiology of inflammation-based disease, as well.  However, there is still limited data on the efficacy of different doses of LPS in inducing inflammation in RAW 264.7 cells. This study aimed to evaluate the effect and safety of LPS at various doses in RAW 264.7 cells. RAW 264.7 cells were exposed to LPS at different dose ranges (10ng/mL-10µg/mL) for 24 hours. The nitric oxide (NO) release as inflammatory responses and viability test were evaluated using Griess assay and CCK-8 assays, respectively. The result showed that NO production was increased at different doses of LPS compared to the control although not significant. Whereas, All LPS-treated RAW 264.7 cells tended to increase but not significantly compared to the control groups. This study showed that the LPS treatment effectively induced inflammation in RAW 264.7 cells as shown by NO production and was considerably safe as the viability was comparable between LPS and control group for RAW cells 264.7 at least up to 10µg/mL for 24 hours.
Anticancer Potential of Cyanidin and Cyanidin-3-Glucoside Through TrkB Receptor Inhibition: Evidence From Molecular Interaction Docking Kurnianingsih, Nia; Suprapto, Ratih Paramita; Ratnawati, Retty
JSMARTech: Journal of Smart Bioprospecting and Technology Vol. 5 No. 2 (2024): JSMARTech Volume 5, No. 2, 2024
Publisher : JSMARTech

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jsmartech.2024.005.02.41

Abstract

Cancer remains a global health problem with persisten demand on therapeutic development to inhibit cancer cells growth without harmful effects on healthy cells. Plant bioactive compounds are intensively studied as anticancer via several signalling pathway. Anticancer therapy via inhibition of tropomyosin kinase receptor-B (TrkB) signal was previously proposed as target therapy. The role of plant metabolites cyanidin and cyanidin-3-glucoside as TrkB inhibitor is has not been investigated. This study was aimed to identify physicochemical of cyanidin and cyanidin-3-glucoside as well as determine it’s role towards TrkB receptor signalling pathway through in silico approach. Molecular docking was performed using Hex 8.0.0 software and visualizes using Discovery Studio Visualizer. The energy binding of TrkB with cyanidin and cyanidin-3-glucoside was -263,21 kcal/mol and 281,65 kcal/mol respectively. Complex of cyanidin-3-glucoside had hydrogen and hydrophobic bond more than TrkB-cyanidin complex. The hydrogen bond formed at Lys637, Arg558 and Gly 561 amino residues. Physicochemical analysis demonstrated that both ligand are potential as kinase enzyme inhibitor. Cyanidin-3-glucoside was predicted more potential as anticancer than cyanidin via TrkB receptor inhibition. Future studies are required to confirm current finding both in-vitro and in-vivo models.
Anthocyanin-Rich Extract from Purple Sweet Potatoes Modified Body Weight, Visceral Fat and Circulatory IL-10 in Stressed-Mice Kurnianingsih, Nia; Rahma, Oktivani Adelathifa; Prayogo, Nabila Putri; Rayhanna, Qanitha Ailsya; Fahanani, Agwin Fahmi; Suprapto, Ratih Paramita; Prakosa, Ardani Galih; Ratnawati, Retty
Pharmaceutical Sciences and Research Vol. 11, No. 3
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Stress-induced inflammation and oxidative stress mechanisms thus plant bioactive compounds as anti-inflammation and antioxidants are placed as novel strategies to resist the adverse effects of stress. Purple sweet potatoes (PSP) as a rich source of macronutrients and anthocyanin showed health benefits as neuroprotective and metabolic diseases. However, limited research explored the effect of PSP extract on stressed animal models. This study aimed to evaluate the bioactive compound analysis of PSP and the effect on body weight gain (BW), visceral fat (VF), and interleukin-10 (IL-10) of stressed model mice. Adult male BALB/c mice (6-8 weeks) were divided into control (CTRL), and stress (STR), stress+anthocyanin-rich extract (ANC) of PSP dose 10, 20, and 40 mg/kgBW. Stress was exposed as restraint stress for 2 hours/day for 14 days. The ANC was administered once a day orally. The IL-10 was measured by enzyme-linked immunosorbent assay (ELISA). Qualitative plant compound analysis showed the presence of flavonoids, alkaloids, phenolics, glycosides, and tannins in PSP extract. The PSP extract also contains a high level of total anthocyanin, 2468.9 ± 159.38 mg/l, and high antioxidant activity. This study observed tendencies to decrease BW and VF following ANC dose of 10 mg/kgBW treatment. The administration of ANC significantly increased the level of circulatory IL-10 cytokine. In conclusion, an anthocyanin-rich extract of PSP modified BW, VF, and IL-10 levels of restraint-stressed model mice.
The Effect of Lipopolysaccharide Challenge in RAW 264.7 Cells on Nitric Oxide Release and Cell Viability Suprapto, Ratih Paramita; Kusumastuty, Inggita; Rizal, Ardian; Adi Nugroho, Dwi
Jurnal Kedokteran Brawijaya Vol. 33 No. 2 (2024)
Publisher : Fakultas Kedokteran Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jkb.2024.033.02.2

Abstract

Lipopolysaccharide (LPS) is a major component of a gram-negative bacterial wall that is widely used and well-established to induce inflammation in vitro. In addition, the in vitro model using RAW 264.7 cells is the most commonly applied in screening the anti-inflammatory and elucidating the pathophysiology of inflammation-based disease, as well.  However, there is still limited data on the efficacy of different doses of LPS in inducing inflammation in RAW 264.7 cells. This study aimed to evaluate the effect and safety of LPS at various doses in RAW 264.7 cells. RAW 264.7 cells were exposed to LPS at different dose ranges (10ng/mL-10µg/mL) for 24 hours. The nitric oxide (NO) release as inflammatory responses and viability test were evaluated using Griess assay and CCK-8 assays, respectively. The result showed that NO production was increased at different doses of LPS compared to the control although not significant. Whereas, All LPS-treated RAW 264.7 cells tended to increase but not significantly compared to the control groups. This study showed that the LPS treatment effectively induced inflammation in RAW 264.7 cells as shown by NO production and was considerably safe as the viability was comparable between LPS and control group for RAW cells 264.7 at least up to 10µg/mL for 24 hours.