Chemical profile of Roselle extract and its inhibitory activities on three digestive enzymes in vitro and in vivo.

Roselle is rich in an extensive diversity of beneficial substances, including phenolic acids, amino acids, anthocyanins, vitamins, and flavonoids. Herein, the chemical constituents in Roselle extract (RE) were identified by UPLC-DAD-QTOF-MS. Besides, its inhibitory effects on three digestive enzymes, i.e. α-amylase, α-glucosidase, and pancreatic lipase, were investigated in both in vitro and in vivo. Thirty-three constituents including hibiscus acid, 18 phenolic acids, 2 anthocyanins and 12 flavonoids were identified. The anthocyanins content in RE was 21.44 ± 0.68 %, while the contents of chlorogenic acids, rutin and quercetin were 17.76 ± 2.28 %, 0.31 ± 0.01 % and 0.32 ± 0.01 %, respectively. RE inhibited pancreatic lipase in a non-competitive way with an IC50 value of 0.84 mg/mL. Besides, it demonstrated a mixed-type inhibition on both α-glucosidase and α-amylase with IC50 values of 0.59 mg/mL and 1.93 mg/mL, respectively. Fluorescence quenching assays confirmed the binding of RE to the enzyme proteins. Furthermore, rats pre-treated with RE at doses of 50 and 100 mg/kg body weight (bwt) exhibited significant reductions in fat absorption and improvements in fat excretion through feces. Additionally, the in vivo study revealed that RE was effective in suppressing the increase of blood glucose after starch consumption, while its effects on maltose and sucrose consumption were relatively weak.

In vitro and in vivo Inhibitory Activity of C-glycoside Flavonoid Extracts from Mung Bean Coat on Pancreatic Lipase and α-glucosidase.

Mung bean is a kind of legume commonly eaten by human. In the present study, a HPLC method for analyzing of two C-glycoside flavonoids, isovitexin and vitexin, in Mung bean was developed. Results showed that the flavonoids are mainly existed in Mung bean coat (MBC), while kernel contains very trace. The extraction of C-glycoside flavonoids from MBC was optimized. MBC extracts with isovitexin and vitexin contents of 29.0 ± 0.28% and 35.8 ± 0.19% were obtained with yield of 1.6 ± 0.21%. MBC extracts exhibited inhibitory activities on pancreatic lipase and α-glucosidase with IC50 values of 0.147 mg/ml and 0.226 mg/ml, respectively. The inhibitory kinetics revealed that MBC extracts showed mixed-type inhibition on these enzymes. Fluorescence quenching titration confirmed the binding of MBC extracts with the enzyme proteins. In vivo study revealed that pre-administration with MBC extracts significantly reduced the triglyceride absorption. Furthermore, it also improved postprandial hyperglycemia in rats through the inhibition of α-glucosidase.

Biocatalytical Acyl-Modification of Puerarin: Shape Gut Microbiota Profile and Improve Short Chain Fatty Acids Production in Rats.

Gut microbiota balance and metabolites have become a potentially mechanism in maintaining health. The specific aim of this study was to compare the modulation of puerarin and puerarin acid esters on gut microbial composition and metabolites. Male mice were fed a control diet or diets supplemented with puerarin, puerarin propanoate ester, puerarin hexanoate ester, puerarin myristate ester for 24 h, respectively. The result revealed that puerarin acid esters with different chain lengths showed different activities to create more own impacted bacterial. Puerarin propanoate and puerarin hexanoate ester significantly improved the diversity of microbiota and promoted the relative abundance of beneficial gut microbiota such as Lactobacillus, Barnesiella, Clostridium IV, Prevotella. Additionally, the puerarin propanoate ester group showed the capacity to deliver specific propionic acid to the colon. But esters with medium-long chain lengths had more opportunity to alter gut microbiota for enhancing the short chain fatty acids production. As a whole, puerarin acid esters with different chain lengths supplements shaped different gut microbial and short chain fatty acids metabolism, which could improve human health.

Total flavone of Abelmoschus manihot suppresses epithelial-mesenchymal transition via interfering transforming growth factor-ß1 signaling in Crohn's disease intestinal fibrosis.

AIM: To explore the role and mechanism of total flavone of Abelmoschus manihot (TFA) on epithelial-mesenchymal transition (EMT) progress of Crohn's disease (CD) intestinal fibrosis. METHODS: First, CCK-8 assay was performed to assess TFA on the viability of intestinal epithelial (IEC-6) cells and select the optimal concentrations of TFA for our further studies. Then cell morphology, wound healing and transwell assays were performed to examine the effect of TFA on morphology, migration and invasion of IEC-6 cells treated with TGF-ß1. In addition, immunofluorescence, real-time PCR analysis (qRT-PCR) and western blotting assays were carried out to detect the impact of TFA on EMT progress. Moreover, western blotting assay was performed to evaluate the function of TFA on the Smad and MAPK signaling pathways. Further, the role of co-treatment of TFA and si-Smad or MAPK inhibitors has been examined by qRT-PCR, western blotting, morphology, wound healing and transwell assays. RESULTS: In this study, TFA promoted transforming growth factor-ß1 (TGF-ß1)-induced (IEC-6) morphological change, migration and invasion, and increased the expression of epithelial markers and reduced the levels of mesenchymal markers, along with the inactivation of Smad and MAPK signaling pathways. Moreover, we revealed that si-Smad and MAPK inhibitors effectively attenuated TGF-ß1-induced EMT in IEC-6 cells. Importantly, co-treatment of TFA and si-Smad or MAPK inhibitors had better inhibitory effects on TGF-ß1-induced EMT in IEC-6 cells than either one of them. CONCLUSION: These findings could provide new insight into the molecular mechanisms of TFA on TGF-ß1-induced EMT in IEC-6 cells and TFA is expected to advance as a new therapy to treat CD intestinal fibrosis.

[Ultracytochemical localization of calcium and ATPase activity on the 2,4-D induced somatic embryogenesis of Lycium barbarum L].

In order to research the function mechanism of the 2,4-D during the development of plant somatic embryogenesis, we studied its function mechanism and relationship with the space-time distributing of Ca2+ content and ATPase activity on somatic embryogenesis of Lycium barbarum L. The possible effects on 2,4-dichlorophenoxyacetic acid (2,4-D) induced somatic embryogenesis and changes of Ca2+ and ATPase active at different development period of somatic embryogenesis. The result showed: The 2,4-D was a key hormone for induced embryonic state of Lycium barbarum L. The embryonic callus and non-embryonic callus was separately obtained in the medium that contains the auxin 2,4-D and lack 2,4-D. In the present study, we have observed the Ca2+ was more abundant in the further intercellular matrix and on the cell wall at the multi-cellular stage, and Ca2+ was concentrated in the plasma membrane and vacuoles membrane during embryonic cell differentiate and division, to the globular embryo, more Ca2+ was seen in the nucleus. Afterward, it was also observed to be distributed in the thicken cell wall and intercellular matrix. At the same process, the variations of ATPase activity and Ca2+ were highly similar, ATPase activity was mainly located on the plasma membrane in early embryogenic cells. With further development, it was also observed to be distributed in endoplasm, nucleus and vacuoles, with the thickening of embryogenic cell wall, ATPase activity was found in the thickened region and the intercellular space. However, the variations of ATPase activity and Ca2+ have not clearly observed variety dynamics at the nonembryogenic callus, and with further vacuolation of nonembryogenic cell, Ca2+ content and ATPase activity gradually drop. It was indicated there was a closely relationship between the dynamics of Ca2+ and ATPase activity in somatic embryogenesis by 2,4-D induced. And the space-time distribution of Ca2+ and ATPase activity play a key role on signal transmission and the regulation of relevant gene expression.