ABSTRACT
Objective: Based on response surface methodology, HPLC was applied to quantitatively determine the optimal processing technology of Glycyrrhizae Radix et Rhizoma Praeparata cum Melle (GRRPM) from the perspective of multi-index and comprehensive evaluation. Methods: HPLC was used for quantitative analysis, and the content of liquiritin, liquiritigenin, licochalcone A and glycyrrhetinic acid was used as inspection indexes. Response surface methodology was used to investigate the effects of the adding amount of honey, steaming and soaking time, frying temperature and frying time on the processing technology of GRRPM, and to optimize the optimal processing technology of GRRPM. Results: The chromatographic column was Diamonsil C18 (2) (4.6 mm × 200 mm, 5 μm); mobile phase was acetonitrile-0.1% phosphate aqueous solution, gradient eluting: 0-20 min, 12%-32% acetonitrile; 20-45 min, 32%-70% acetonitrile; 45-75 min, 70%-97% acetonitrile, with detection wavelength of 260 nm, column temperature of 20 ℃, and flow rate of 1 mL/min; Using liquiritin as internal standard, the relative correction factors of glycyrrhizin, licochalcone A, glycyrrhizinic acid and their relative correction factors were determined and calculated to be 0.56, 0.64 and 1.42, respectively. The optimum processing process of GRRPM was as follows: the amount of honey was 1/4, the soaking time was 15 min, frying pan bottom temperature was 160 ℃, and frying time was 13 min. Conclusion: The results of systematic adaptability investigation of the experimental content determination method meet the requirements. The best processing scheme of GRRPM optimized by response surface methodology is feasible and provides scientific basis for formulating quality standards and modern research of GRRPM.
ABSTRACT
Resveratrol (3,5,4'-trihydroxystilbene, RSV) has been widely used in mammalian cells, but whether it can be used during freezing boar semen is still unknown. The effects of RSV treatment during boar semen freezing on its anti-freezing ability, apoptosis, and possible apoptotic pathways were observed in this study. Sperm motility, mitochondrial membrane potential (ΔΨ), adenosine triphosphate (ATP) content, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL)-positive apoptotic state, and messenger RNA (mRNA) expression levels of apoptotic genes involved in different apoptotic pathways after freezing with or without RSV treatment were tested. The results showed that: (1) Compared with fresh sperm, the motility, normal acrosome rate, and plasma membrane integrity rate of frozen boar sperm decreased significantly (P0.05), but it did significantly improve the normal acrosome rate (57.65% vs. 47.00%, P<0.05) and plasma membrane integrity rate (46.67% vs. 38.85%, P<0.05). (2) After freezing, most boar sperm showed low mitochondrial ΔΨ. RSV treatment could increase the rate of high mitochondrial ΔΨ of boar sperm. (3) RSV treatment significantly decreased reactive oxygen species (ROS) levels (58.65% vs. 88.41%, P<0.05) and increased the ATP content (0.49 μmol/L vs. 0.25 μmol/L, P<0.05) of boar sperm during freezing. (4) The apoptotic rate of the freezing group (80.41%) with TUNEL detection increased significantly compared to the fresh group (9.70%, P<0.05), and RSV treatment greatly decreased the apoptotic rate (68.32%, P<0.05). (5) Real-time polymerase chain reaction (RT-PCR) showed that not only the genes from the death receptor-mediated apoptotic pathway (tumor necrosis factor-α (TNF-α), Fas ligand (FasL), and Caspase-8), but also the genes from the mitochondria-mediated apoptotic pathway (manganese superoxide dismutase (MnSOD), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and Caspase-9) were both significantly changed after freezing. RSV treatment during freezing greatly changed their expression levels. Although RSV treatment during boar semen freezing did not significantly increase motility after thawing, it still played an efficient antioxidant role, which could enhance the mitochondrial function and decrease the apoptotic level induced by both the death receptor- and mitochondria-mediated apoptotic pathways.
ABSTRACT
Objective: To establish the HPLC fingerprint and multicomponents determination of Potentillae Chinensis Herba, and provide a scientific basis for the improvement of its quality standards. Methods: The separation was performed on a chromatographic Diamonsil C18 column (250 mm × 4.6 mm, 5 μm), with acetonitrile-0.1% phosphoric acid as the mobile phase for gradient elution. Volume flow rate was 1.0 mL/min. Column temperature was 25 ℃. Injection was 10 μL and the detection wavelength was 260 nm. The fingerprints of 10 batches of Potentillae Chinensis Herba were established and evaluated by the similarity evaluation system of TCM (version 2004A), which were divided into two categories by clustering analysis. Meanwhile, the content of galic acid, quercetin, and kaempferol was determined. Results: The fingerprint of Potentillae Chinensis Herba was established. There were 12 common peaks in the fingerprint. Galic acid, quercetin, and kaempferol were separated with good linearity relationships (r > 0.999). The average recovery rates of the investigated compounds were 97.44%, 97.64%, and 99.19%, respectively. Conclusion: The established fingerprint and multicomponents determination method of Potentillae Chinensis Herba have strong specificity and good repeatability, which can effectively control the internal quality of Potentillae Chinensis Herba and provide reference for improving the quality evaluation method of Potentillae Chinensis Herba.
ABSTRACT
Diabetes mellitus (DM) is a metabolic disease characterized by hyperglycemia, with a high incidence and many complications. It has become an increasingly serious public health problem in the world, and has seriously affected the quality of life. Phosphatidylinositol-3-kinases (PI3K)/protein kinase B (Akt) signaling pathway is the main pathway of insulin signal transmission and the main signal channel for regulating blood glucose. The abnormal signal molecule of PI3K/Akt may cause abnormal signal transduction pathway, so as to impact the proliferation, apoptosis, metastasis and invasion of the corresponding tissues and organs, and lead to the occurrence of disease. Study of PI3K/Akt signal channel has a positive significance for investigating whether traditional Chinese medicine(TCM) has a definite and stable hypoglycemic effect. Currently, there are many TCM and Western medicines to treat diabetes, however, most drugs, especially Western medicines, have a relatively poor effect in controlling complications. To understand the progress of TCM in treatment of diabetes, in expectation of better studying the comprehensive therapeutic effect and mechanism of TCM on diabetes, and further developing the multi-target, multi-way and multi-channel advantages and features of TCM in the treatment of diabetes, this paper focuses on a systematic analysis on the progress of in vivo and in vitro studies on DM based on PI3K/Akt signaling channel in recent years, including the effect of the signaling channel on insulin secretion, the three main target organs of insulin (liver, skeletal muscle and fat), and its effect on the four main complications of diabetes (brain, kidney, heart, testis), and also provides certain ideas and guidance for the study of hypoglycemic mechanism of TCM monomer, TCM and compound medicine.