Identification of Pheophytin a and Hydroxy Pheophytin a from Rang Chuet (Thunbergia laurifolia Linn.) as Potent NQO-1 Inducers in Liver Cells.

Thunbergia laurifolia Linn. (Rang Chuet, RC), a Thai medicinal plant, possesses various bioactive compounds with potential health benefits. This study aimed to identify detoxifying compounds within RC crude extract. RC leaves were extracted using the Soxhlet method with chloroform. Total carotenoids, chlorophylls, extract yield, total phenolic contents (TPCs), and total flavonoid contents (TFCs) were measured. The extract's composition was analyzed. Cytotoxicity and effects on the detoxification enzyme NQO-1 were assessed in liver cell lines (AML12 and HepG2) using MTT and NQO-1 assays, respectively. Bioactive fractions were identified using fractionation techniques and mass spectrometry (LC-MS). RC extract displayed significant levels of carotenoids (0.375 mg/g), chlorophylls (2.682 mg/g), and favorable yield (15.3%). TPC and TFC were 363.776 mg/g and 112.22 mg/g of extract, respectively. Analysis revealed phenolic acids (gallic acid, caffeic acid), flavonoid (apigenin), chlorophylls (chlorophylls a, b, pheophytin a and b), and lutein. Among the fractions, Fraction 3 (F3) exhibited the highest NQO-1 enzyme activity. F3 contained pheophytin a and hydroxy pheophytin a, confirmed by LC-MS (m/z 871.59+ [M + H]+ and 887.59+ [M + H]+). F3 significantly induced NQO-1 activity in both HepG2 (3.908-fold) and AML12 (1.99-fold) cells. This study identified F3 from RC extract as a promising fraction containing pheophytin a and hydroxy pheophytin a, responsible for inducing the detoxification enzyme NQO-1 in liver cells. These findings suggest RC's potential for promoting detoxification.

The Microstructure, Rheological Characteristics, and Digestibility Properties of Binary or Ternary Mixture Systems of Gelatinized Potato Starch/Milk Protein/Soybean Oil during the In Vitro Digestion Process.

The in vitro digestibility of potato starch-based foods interacting with milk protein and soybean oil was investigated. Microstructures and rheological changes upon digestion were determined. The results showed that the addition of milk proteins (casein and whey protein) promoted gelatinized potato starch digestion, while soybean oil slowed down gelatinized potato starch digestion. A mixture of soybean oil and milk protein promoted the digestion of milk protein, while a mixture of gelatinized potato starch and milk protein inhibited the digestion of milk protein. The mixture of milk protein and/or gelatinized potato starch with soybean oil promoted the release of free fatty acids in soybean oil. The highest release rate of free fatty acids was attained by a mix of milk protein and soybean oil. The mixed samples were digested and observed with a confocal laser scanning microscope. The viscosity of the digestates was determined by a rheometer. Overall, the results demonstrated that the addition of milk protein and soybean oil had an effect on the in vitro digestibility of gelatinized potato starch and its microstructure.

Evaluation of the physico-chemical properties of potato starch-based foods and their interactions with milk protein and soybean oil.

Physico-chemical properties of potato starch-based foods (PSBF) interacted with milk protein (MP), and soybean oil (SBO) were investigated. Microstructures, rheological properties, and chemical bonding among those ingredients were determined. An emulsion-filled gel, in which oil droplets stabilized by MP and/or amylose-lipid complex (ALC) dispersed in a starch gel structure of PSBF was revealed by confocal laser scanning microscopy. Starch-starch, protein-oil, and protein-protein played the dominant interactions while ALC and starch-protein interaction were subordinates. Rheological data showed that MP induced a thinning effect on starch gel, while SBO seemed to reinforce the solid-like properties of the gel. The chemical interactions analyzed by FTIR, Raman, and X-ray diffraction suggested that these foods were lack in non-covalent crosslinks and were dominated by diverse physical interactions. However, the different preparation of such foods could induce chemical binding in a different way and MP and SBO could affect the properties of PSBF in this study.

The Influence of Whey Protein Heating Parameters on Their Susceptibility to Digestive Enzymes and the Antidiabetic Activity of Hydrolysates.

The inhibition of dipeptidyl peptidase-IV (DPP-IV) and the release of glucagon-like peptide-1 (GLP-1) could normalize blood glucose levels in diabetic patients. This study evaluated the susceptibility of whey proteins to enzyme hydrolysis and the antidiabetic properties of protein hydrolysates from ß-lactoglobulin (ß-LG) and α-lactalbumin (α-LA) solutions compared with whey protein isolate (WPI) solution treated at different heating temperatures (65, 75, and 85 °C). α-LA hydrolysate provided the lowest degree of hydrolysis (DH). Those heating temperatures did not significantly affect the DH of all protein hydrolysates. α-LA hydrolysate significantly increased GLP-1 levels and DPP-IV inhibitory activity more than ß-LG hydrolysate. WPI hydrolysate inhibited DPP-IV activity less than an α-LA hydrolysate, but they were no significant differences for GLP-1 release activity. Heat treatment could affect the antidiabetic properties of all protein hydrolysates. Heating at 75 °C resulted in greater inhibition of the activity of DPP-IV than at 65 and 85 °C. The highest increase in GLP-1 release was also observed by heating at 75 °C. The recently obtained information is useful for the utilization of α-LA, heated at 75 °C for 30 min, in the preparation of antidiabetic food supplements.