Abstract

BackgroundTempeh is a widely known fermented soybean that contains elevated level of bioactive contents. Our previous study has shown that anaerobic fermented Nutrient Enriched Soybean Tempeh (NESTE) with increase amino acid and antioxidant levels possessed better hepatoprotective effect than raw soybean.MethodsIn this study, the anti-inflammatory effect of the NESTE aqueous extract and raw soybean aqueous extract (SBE) were evaluated by quantifying the inhibition of IL-1β, TNF-α and nitric oxide (NO) secretion in LPS treated RAW 264.7 cell in vitro. On the other hand, in vivo oral acute toxicity effect of the extract was tested on mice at the dose of 5000 mg/kg body weight. In vivo oral analgesic effect of both aqueous extracts at 200 and 1000 mg/kg body weight was evaluated by the hot plate test.ResultsIn the in vitro anti-inflammatory study, 5 mg/mL NESTE was able to inhibit 25.50 ± 2.20%, 35.88 ± 3.20% and 28.50 ± 3.50% of NO, IL-1β and TNF-α production in LPS treated RAW 264.7 cells without inducing cytotoxic effect on the cells. However, this effect was lower than 4 μg/mL of curcumin, which inhibited NO, IL-1β and TNF-α production by 89.50 ± 5.00%, 78.80 ± 6.20% and 87.30 ± 4.00%, respectively. In addition, 1.5 to 2.5-fold increase of latency period up to 120 min for mice in the hot plate test was achieved by 1000 mg/kg NESTE. The analgesic effect of NESTE was better than 400 mg/kg of acetyl salicylic acid, which only increased ~ 1.7-fold of latency period up to 90 min. Moreover, NESTE did not show acute toxicity (no LD50) up to 5000 mg/kg body weight.ConclusionNESTE is a nutritious food ingredient with potential anti-inflammatory and analgesic effects.

Highlights

  • Tempeh is a widely known fermented soybean that contains elevated level of bioactive contents

  • Both Nutrient Enriched Soybean Tempeh (NESTE) and Soybean aqueous extract (SBE) did not induce cytotoxic effect toward RAW 264.7 cell even at the highest concentration of 5 mg/mL. Both extracts have no significant differences with the control group. This result indicates that both NESTE and SBE did not influence the viability of RAW 264.7 cell

  • 5 mg/mL of NESTE showed the highest inhibitory effect, ie 25.50 ± 2.20%, 35.88 ± 3.20% and 28.50 ± 3.50%, respectively on nitric oxide (NO), IL-1β and TNF-α production by LPS treated RAW 264.7 cells as compared to 5 mg/mL of SBE (18.50 ± 2.0%, 27.80 ± 3.40% and 20.00 ± 2.30%, respectively), which indicated that fermentation process has significantly improved the in vitro antiinflammatory effect of soybean

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Summary

Introduction

Tempeh is a widely known fermented soybean that contains elevated level of bioactive contents. Our previous study has shown that anaerobic fermented Nutrient Enriched Soybean Tempeh (NESTE) with increase amino acid and antioxidant levels possessed better hepatoprotective effect than raw soybean. Macrophage is one of the immune cells that generate the chemical mediators such as reactive oxygen species (ROS) and reactive nitrogen species (RNS) when the inflammatory signalling response is activated by immune-stimulants, including bacterial endotoxin Active metabolites such as proteins, phenolic compounds and antioxidants that were found in the soybean-based. One of the most commonly consumed fermented soybeans around South East Asia is tempeh which contained significant level of antioxidants and amino acids [12, 13], which may subsequently contribute to the anti-inflammatory effect.

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