Inhibitory Effect of Persimmon Tannin on Pancreatic Lipase and the Underlying Mechanism in Vitro

Pancreatic lipase (PL) is a critical enzyme associated with hyperlipidemia and obesity. A previous study of ours suggested that persimmon tannin (PT) was the main component accounting for the antihyperlipidemic effects of persimmon fruits, but the underlying mechanisms were unclear. In this present study, the inhibitory effect of PT on PL was studied and the possible mechanisms were evaluated by fluorescence spectroscopy, circular dichroism (CD) spectra, isothermal titration calorimetry (ITC), and molecular docking. PT had a high affinity to PL and inhibited the activity of PL with the half maximal inhibitory concertation (IC₅₀) value of 0.44 mg/mL in a noncompetitive way. Furthermore, molecular docking revealed that the hydrogen bonding and π–π stacking was mainly responsible for the interaction. The strong inhibition of PT on PL in the gastrointestinal tract might be one mechanism for its lipid-lowering effect

Structure-Dependent Membrane-Perturbing Potency of Four Proanthocyanidin Dimers on 3T3-L1 Preadipocytes

Proanthocyanidins (PAs) have been widely recognized for their broad spectrum of beneficial health effects, which are highly structure-dependent. It was found that PA dimers epicatechin-3-gallate-(4β→8,2β→O→7)-epicatechin-3-gallate (A-type ECG dimer) and epigallocatechin-3-gallate-(4β→,2β→O→7)-epigallocatechin-3-gallate (A-type EGCG dimer) inhibit the differentiation of 3T3-L1 cells significantly, whereas epicatechin-(4β→8,2β→O→7)-epicatechin (A-type EC dimer) and epicatechin-(4β→8)-epicatechin (B-type EC dimer) showed little effect in previous work. However, the underlying mechanisms are unclear. To test whether bilayer perturbation may underlie this diversity of actions, we examined the bilayer-modifying effects of the four dimers in both 3T3-L1 cell and 1,2-dipalmitoyl-<i>sn</i>-glycero-3-phosphocholine liposome models by using scanning electron microscopy, fluorescent spectroscopy, differential scanning calorimetry, and molecular dynamics methods. Our results revealed that A-type ECG and EGCG dimers had a high affinity for the lipid bilayer and could form simultaneous hydrogen bonds (H-bond) with both the surface oxygen acceptors and the deeper inside lipid oxygen atoms. However, A-type and B-type EC dimers contacted only the surface oxygen atoms with limited and significantly fewer H-bonds. A-type ECG and EGCG dimers notably distorted the membrane morphology of 3T3-L1 cells. In the present study, we found there was a high positive correlation between the membrane-disturbing abilities of the four dimers and their 3T3-L1 cell differentiation inhibitory effects as previously reported. This indicated that the strong 3T3-L1 cell differentiation inhibitory effect of A-type ECG and EGCG dimers might be due to their strong bilayer-perturbing potency