Inhibitory kinetics and bioactivities of Nuciferine and Methyl Ganoderate on Mucor miehei lipase and 3T3-L1 preadipocytes.

In this study, inhibitory kinetics of Nuciferine and Methyl Ganoderate extrated from Lotus Leaves and Ganoderma lucidum on Mucor miehei Lipase were studied first. The molecular structure of Nuciferine and Methyl Ganoderate were determined. The inhibitory effects of two extracts on lipase were reversible, with the IC50 values of 0.194 and 0.332 mg/mL, respectively. The inhibition kinetic analysis by Lineweaver-Burk plots showed that they were a mixed-type inhibitor of lipase, with inhibition constants KI of 0.16 and 0.29 mg/mL, and KIS of 0.36 and 0.49 mg/mL, respectively. Results of spectral analysis showed that the UV absorption and the molecule fluorescence spectrum of the lipase hydrolyzate were significantly decreased after the inhibitor was added. The molecular docking further suggested that the interaction site between the active substance and inhibitor was located in an α-helix and a ß-sheet of the lipase, and the lipase active site was interfered by the inhibitor near the cap structure. In addition, the proliferation and differentiation of 3 T3-L1 preadipocytes were inhibited by two extracts. Total triglycerides and cholesterol were significantly reduced in the cells. The results confirmed that Nuciferine and Methyl Ganoderate can be used as potential obesity treatment drugs.

[Study on UHPLC fingerprint and determination of eight phenolic components of Tetrastigma hemsleyanum leaves].

A novel method combining ultra-high performance liquid chromatography (UHPLC) fingerprint and simultaneous quantitative analysis of eight phenolic components was developed and validated for quality evaluation of Tetrastigma hemsleyanum leaves. For fingerprint analysis, 15 peaks were selected as the common peaks to evaluate the similarities among 41 batches of T. hemsleyanum leaves collected from different regions. Additionally, simultaneous quantification of eight markers, including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isoorientin, orientin, vitexin-2-O-rhamnoside,vitexin and isovitexin, was performed and the obtained data demonstrated that our method has achieved desired linearity, precision and accuracy. Clustering statistical analysis was further application in T. hemsleyanum leaves from different regions. The results indicated that new approach conbine ultra-high performance liquid chromatography (UHPLC) fingerprint and simultaneous quantitative analysis of eight phenolic components was applicable in quality control of T. hemsleyanum leaves.

[Differential protein analysis on the root response of rice with high phosphorous uptake efficiency to low phosphorous stress].

A comparative proteomics analysis was performed to identify the molecular response of a rice cultivar (Oryza sative cv. 'IRRI71331') with high phosphorous (P) uptake efficiency to low P stress. The hydroponically grown rice plants were provided with two levels of P (0.5 mg x L(-1) and 10 mg x L(-1)) supplied in quarter strength Kimura solution, and the root total proteins extracted on the 3rd and 6th day of treatments were separated by two-dimensional gel electrophoresis (2-DE). Comparing with the control (10 mg x L(-1) of P), a total of 29 protein spots under low P stress (0.5 mg x L(-1)) showed differences in their relative abundance, among which, 17 were higher, 11 were lower, and 1 was novel on the 3rd day, and 8 were induced, 19 were suppressed, 1 was disappeared, and 1 had no obvious change on the 6th day. Ten differentially expressed protein spots were identified by MALDI-TOF/MS, and searched in protein databases. According to the putative functions, the identified proteins were classified into four groups, i.e., signal transduction (glycine-rich RNA-binding protein, phosphate starvation response regulator-like), gene expression (putative pre-mRNA splicing factor, putative AAA-metalloprotease), metabolism (adenylosuccinate lyase, serpin, S-adenosylmethionine synthetase, MYB transcription factor-like protein), and ion transport (cation-transporting ATPase, sarcoplasmic reticulum protein). The identified proteins were involved in various physiological responses to enhance stress resistance, such as signal recognition and transduction, RNA cleavage, degradation of denatured protein, and ion transportation and cellular ion balance. The serine protease inhibitor and S-adenosylmethionine synthetase and the MYB transcription factor-like protein, which were the key proteins associated with P deficiency--tolerance of other species, were affected by the same stress for rice. The results indicated that the tolerance to low P stress was controlled by a complex signal transduction and metabolism regulation network in rice root system.