Sub-chronic toxicity study of green peas protein hydrolysate in rats.

Despite green peas protein hydrolysate benefits, very few studies have described the potential toxicity. The acute oral toxicity of green peas protein which hydrolyzed by bromelain at a single dose of 5000 mg/kg BW has been evaluated in Swiss Webster mice and did not produce treatment-related signs of toxicity or mortality in any of the animals tested during the 14-day observation period. The present study aimed was to evaluate the sub-chronic toxicity effects of Protein Hydrolysate Green Peas Bromelain (PHGPB) in Sprague Dawley rats by the regulations of the Indonesian Food and Drug Supervisory Agency 2013. In the repeated dose 28-day oral toxicity study, the administration of 100 mg/kg, 200 mg/kg, and 400 mg/kg/day of green peas protein hydrolysate per body weight revealed no significant difference (P > 0.05) in body weight change, hematological parameters, relative organ weights, and gross findings compared to the control group. Clinical biochemistry analysis and histopathology examinations of liver and kidney showed slight morphological alteration. The oral sub-chronic toxicity test of PHGPB for 28 days did not induce noticeable signs of toxicity. The no-observed adverse effect level (NOAEL) of PHGPB in the sub-chronic toxicity study was dose of the 200 mg/kg BW. The results of our studies PHGPB indicate a lack of toxicity and support the use of functional foods.

Quercetin prevents chronic kidney disease on mesangial cells model by regulating inflammation, oxidative stress, and TGF-ß1/SMADs pathway.

Background: Chronic kidney disease (CKD) happens due to decreasing kidney function. Inflammation and oxidative stress have been shown to result in the progression of CKD. Quercetin is widely known to have various bioactivities including antioxidant, anticancer, and anti-inflammatory activities. Objective: To evaluate the activity of quercetin to inhibit inflammation, stress oxidative, and fibrosis on CKD cells model (mouse mesangial cells induced by glucose). Methods and Material: The SV40 MES 13 cells were plated in a 6-well plate with cell density at 5,000 cells/well. The medium had been substituted for 3 days with a glucose-induced medium with a concentration of 20 mM. Quercetin was added with 50, 10, and 5 µg/mL concentrations. The negative control was the untreated cell. The levels of TGF-ß1, TNF-α, and MDA were determined using ELISA KIT. The gene expressions of the SMAD7, SMAD3, SMAD2, and SMAD4 were analyzed using qRT-PCR. Results: Glucose can lead to an increase in inflammatory cytokines TNF-α, TGF-ß1, MDA as well as the expressions of the SMAD2, SMAD3, SMAD4, and a decrease in SMAD7. Quercetin caused the reduction of TNF-α, TGF-ß1, MDA as well as the expression of the SMAD2, SMAD3, SMAD4, and increased SMAD7. Conclusion: Quercetin has anti-inflammation, antioxidant, antifibrosis activity in the CKD cells model. Thus, quercetin is a promising substance for CKD therapy and further research is needed to prove this in CKD animal model.

Detam 1 black soybean against cisplatin-induced acute ren failure on rat model via antioxidant, antiinflammatory and antiapoptosis potential.

Background and aim: Cis-Diamminedichloroplatinum (II) (Cisplatin) is one of the most synthetic anticancer drug but have several adverse effects and one of them is acute ren failure. Cisplatin can induce nephrotoxicity occur via the toxic generation of reactive oxygen species (ROS). Black soybean (Glycine max L. Merr.) has been reported contain high levels of phenolics and anthocyanins that has antioxidant activity. This study aims to determine the effect of ethanol extract of black soybean (EEBS) against cisplatin-induced nephrotoxicity in rats. Experimental procedure: Cisplatin-induced nephrotoxicity rats treated with EEBS and the blood samples taken on days 0, 9, and 18. The effects of EEBS was evaluated by determining Interferon-γ (IFN-γ), Caspase-3 (Casp-3), and Interleukin-1ß (IL-1ß) expression using immunohistochemistry (IHC), blood urea nitrogen (BUN), Uric Acid (UA) content and catalase (CAT) content in the blood plasma with colorimetric assay kit. Results and conclusion: Based on the results, EEBS treatment had successfully reduced pro-inflammatory cytokines IL-1ß and IFN-γ, and improved physiological condition by lowering BUN and UA content while increasing CAT activity. No significant effect was found in Casp-3 expression. EEBS has potential to improve acute renal failure condition through inflammatory suppression and renal function improvement.

Nutrition profile and potency of RGD motif in protein hydrolysate of green peas as an antifibrosis in chronic kidney disease.

OBJECTIVES: Fibrosis is the major cause of chronic kidney injury and the primary etiology in diabetic glomerulosclerosis. The initial study of protein hydrolysate of green peas hydrolyzed by bromelain (PHGPB) considered it to improve kidney function parameters and showed no fibrosis in histopathology features in gentamicin-induced nephrotoxicity rats. In the current study, we aimed to assess the nutrition profile and potency of RGD in PHGPB as antifibrosis in chronic kidney disease (CKD). MATERIALS AND METHODS: Green peas (Pisum sativum) were hydrolyzed by bromelain from pineapple juice to obtain PHGPB. The amino acid content of PHGPB was measured using the UPLC method, while the primary structure used LC-MS/MS. Bioinformatic analysis was conducted using the Protease Specificity Predictive Server (PROSPER). The potency of RGD in PHGPB was characterized by determining the levels of Fibronectin (FN) and TGF-ß1 in mesangial SV40 MES 13 cell lines of diabetic glomerulosclerosis. RESULTS: The level of lysine was 364.85 mg/l. The LC-MS/MS data showed two proteins with 4-15 kDa molecular weight originated from convicilin (P13915 and P13919) which were predicted by PROSPER proteolytic cleavage, resulted in RGD in the LERGDT sequence peptide. PHGPB increased SV40 MES 13 mesangial cell proliferation that died from high-glucose levels (diabetic glomerulosclerosis model). PHGPB and RGD reduced the levels of FN and TGF-ß1 in mesangial cell lines of diabetic glomerulosclerosis. CONCLUSION: The nutrition profile and RGD motif in PHGPB show great potential as antifibrosis in CKD.

Kidney therapeutic potential of peptides derived from the bromelain hydrolysis of green peas protein.

OBJECTIVES: Kidney disease is a global health problem that needs a solution to its therapy. In the previous study, we found that protein hydrolysate of green peas origin of Indonesia hydrolysed by bromelain (PHGPB) showed improve kidney function in cisplatin-induced nephropathy rats. In this study, we investigated the effect of PHGPB to obtain effective dose that exerts a therapeutic effect on chronic kidney disease (CKD) based on reducing urea and creatinine levels and to elucidate its mechanism of action. MATERIALS AND METHODS: Two sets of experiments were conducted: (1) characteristics and proteomic profile of PHGPB, (2) in vivo test of PHGPB in gentamycin-induced Wistar rats, including urea and creatinine measurements, activities of antioxidant and kidney-related peptides (ANP, COX-1, and renin). RESULTS: PHGPB showed three bands under 10 kDa using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and contained 10 identified proteins using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Significant differences in urea and creatinine levels were found between all PHGPB treatments and positive controls (P<0.01). The lowest levels of urea and creatinine that were validated by high super oxide dismutase (SOD) activity and atrial natriuretic peptide (ANP) level were obtained in the 200 mg/day PHGPB treatment. However, the mean renin level was high and cyclooxygenase-1 (COX-1) level did not exceed positive and negative control levels. CONCLUSION: PHGPB at dose 200 mg/kgBW shows a potential CKD therapeutic effect that is dose-dependent. Higher PHGPB dose corresponds to better effect on kidney function by increasing antioxidant activity and ANP levels in gentamycin-induced Wistar rats.