Preparation, identification, activity prediction, and protective effects on IR-HepG2 cells of five novel DPP-IV inhibitory peptides from protein hydrolysate of skipjack tuna dark muscles.

To produce peptides with high dipeptidyl peptidase IV (DPP-IV) inhibitory activity, neutrase was selected from five proteases (trypsin, neutrase, pepsin, alcalase and flavor protease) with the highest degree of hydrolysis (DH) (18.23 ± 1.08%) and DPP-IV inhibitory rate (53.35 ± 4.02%) to produce protein hydrolysate (NPH) from the dark muscles of skipjack tuna (Katsuwonus pelamis). Then, NPH-1 was isolated from NPH by gel permeation chromatography and found to possess the highest DPP-IV inhibitory rate (65.12 ± 7.94% at 0.5 mg ml-1) in the separated components (including NPH-1, NPH-2, NPH-3 and NPH-4). Subsequently, the available prediction models of tripeptides and tetrapeptides with the DPP-IV inhibitory rate were established using an artificial neural network (ANN). The RMSE (0.56 and 0.33 for the model established through collected tripeptides and tetrapeptides, respectively) and R2 (0.95 and 0.99 for the model established through collected tripeptides and tetrapeptides, respectively) of the ANN model's parameters were within acceptable limits, indicating that this model is available. Next, the ANN model was applied to predict tripeptides and tetrapeptides from the hydrolysate of skipjack tuna dark muscles, and five peptides (Ala-Pro-Pro (APP), Pro-Pro-Pro (PPP), Asp-Pro-Leu-Leu (DPLL), Glu-Ala-Val-Pro (EAVP) and Glu-Ala-Iie-Pro (EAIP)) possessing a noticeable DPP-IV inhibitory rate (with DPP-IV IC50 values of 42.46 ± 5.02, 37.71 ± 9.17, 58.85 ± 14.42, 49.94 ± 6.69 and 57.15 ± 6.13 µM, respectively) were screened from the protein hydrolysate. The above five peptides were proved to effectively promote glucose consumption in the insulin resistant-HepG2 (IR-HepG2) cell model considering that the glucose consumption rates of APP, PPP, DPLL, EAVP and EAIP treatment groups are all more than twice that of the dexamethasone group. Accordingly, mechanistic studies showed that these peptides interacted with PI3K/AKT and AMPK signaling pathways and promoted the phosphorylation of PI3K p110, AKT and AMPK (the protein expressions of PI3K p110, p-AKT and p-AMPK in APP, PPP, DPLL, EAVP and EAIP treatment groups are 1.64-2.22 fold compared with that in the dexamethasone group), thereby enhancing glucose uptake and further alleviating insulin resistance. These findings demonstrated that skipjack tuna dark muscle is a potential DPP-IV inhibitory peptide source, and five DPP-IV inhibitory peptides from its hydrolysate may exert potent anti-diabetic activity. In comparison, PPP may be the most potential active ingredient for healthy food against type 2 diabetes mellitus in the five screened peptides considering synthetically the DPP-IV inhibitory rate, bioavailability and synthesis cost.

Heat-Killed Lactobacillus rhamnosus ATCC 7469 Improved UVB-Induced Photoaging Via Antiwrinkle and Antimelanogenesis Impacts.

Exposure of ultraviolet B (UVB) radiation is the main factor from the environment to cause skin photoaging. Lactobacillus rhamnosus ATCC 7469, is a probiotic strain with a good track record for enhancing human health. The present study conducted the impacts of heat-killed L. rhamnosus ATCC 7469 (RL) on photoaging in vitro using mouse skin fibroblast (MSF) cells and human epidermal melanocytes (HEM) exposed to UVB. The results showed that (1) RL-protected UVB-induced cytotoxicity relating to absorb UVB and reduce DNA damage. (2) RL exerted the antiwrinkle impact involved in two aspects. Firstly, RL downregulated MMP-1, 2, 3 expressions associating with MAPK signaling, resulting in the increased the protein expression of COL1A1, further booting type I collagen abundant thereby promoting the antiwrinkle impact in MSF cells. Secondly, RL reduced ROS content, further decreasing oxidative damage relating to Nrf2/Sirt3/SOD2 signaling, thereby promoting the antiwrinkle impact in MSF cells. (3) RL suppressed tyrosinase and TYRP-2 activity and/or levels associating with PKA/CREB/MITF signaling, thereby promoting antimelanogenesis impact in HEM cells. In conclusion, our findings suggest that RL could reduce photoaging caused by UVB via antiwrinkle and antimelanogenesis properties and may be a potential antiphotoaging beneficial component, which is applied in the cosmetic industry.

Heat-Killed Lacticaseibacillus paracasei Ameliorated UVB-Induced Oxidative Damage and Photoaging and Its Underlying Mechanisms.

Ultraviolet B (UVB) radiation is a major environmental causative factor of skin oxidative damage and photoaging. Lacticaseibacillus paracasei is a well-known probiotic strain that can regulate skin health. The present study investigated the effects of heat-killed Lacticaseibacillus paracasei (PL) on UVB linked oxidative damage and photoaging in skin cells (Normal human dermal fibroblast (NHDF) cells and B16F10 murine melanoma cells). Results demonstrated that: (1) PL prevented UVB-induced cytotoxicity relating to decreased DNA damage in NHDF and B16F10 cells; (2) PL alleviated UVB-induced oxidative damage through increasing GSH content, as well as antioxidant enzyme activities and mRNA levels (except MnSOD activity and mRNA levels as well as CAT mRNA level) relating to the activation of Sirt1/PGC-1α/Nrf2 signaling in NHDF cells; (3) PL attenuated UVB-induced photoaging was noticed with a decrease in the percentage of SA-ß-gal positive cells in NHDF cells model. Moreover, PL attenuated UVB-induced photoaging through exerting an anti-wrinkling effect by enhancing the type I collagen level relating to the inhibition (JNK, p38)/(c-Fos, c-Jun) of signaling in NHDF cells, and exerting an anti-melanogenic effect by suppressing tyrosinase and TYRP-1 activity and/or expressions relating to the inhibition of PKA/CREB/MITF signaling in B16F10 cells. In conclusion, PL could ameliorate UVB-induced oxidative damage and photoaging. Therefore, PL may be a potential antioxidant and anti-photoaging active ingredient for the cosmetic industry.