Fourier transform infrared spectroscopic markers of the scorching degree of Scutellariae Radix related to the variation patterns of flavonoids and carbohydrates.

Scutellariae Radix (SR) is a common herb in Asia and Europe. In the clinical practice of traditional Chinese medicine, the raw SR is often stir-baked to partly scorch to reduce the side effects (stomach discomfort, diarrhea, etc.) but enhance some desired effects (such as the hemostatic activity). The scorching degree is crucial to assure the safety and efficacy of the scorched SR. Under-scorching is insufficient to adapt the adverse and favorable activities, while over-scorching can destroy all activities. Up to now, the scorching degree of SR is still determined by the manual observation of colors. Since the visual judgement is vulnerable to personal knowledge and experience, it is difficult to control the optimization and consistence of the scorching degree of SR. This research was designed to explore the potential indicators that can exactly reflect the scorching degree of SR and be measured objectively and quantitatively. A total of 15 morphological and chemical properties as well as Fourier transform infrared (FTIR) spectroscopy were included to profile the variations of the SR slices that were stir-baked at different temperatures for different times. According to the principal component analysis and statistical tests, 10 properties showed close relationships with the scorching degree of SR. As the increase of the heating time at the same heating temperature, the yield, L*, a*, b*, sucrose, baicalin and wogonoside decreased continuously and showed the monotone variation patterns. Meanwhile, 5-HMF, baicalein and wogonin showed the inflection variation patterns, which means these properties increased in the earlier stage but decreased in the later stage during the scorching process. FTIR spectra also revealed the variations of flavonoids and carbohydrates. The scorching degree of SR can be determined by the intuitive observation of FTIR spectral peaks at 1738 cm-1, 1705 cm-1, 1611 cm-1, 1586 cm-1, 1450 cm-1, 1410 cm-1 and 1023 cm-1. In summary, FTIR spectroscopy can be used for the rapid assessment of the scorching degree of SR, while the contents of flavonoid glycosides and aglycones may be used as the quality criteria of the scorched SR.

Luteolin Inhibits the Biofilm Formation and Cytotoxicity of Methicillin-Resistant Staphylococcus aureus via Decreasing Bacterial Toxin Synthesis.

Owing to the fact that luteolin has antibacterial activity against Staphylococcus aureus (S. aureus) and methicillin-resistant S. aureus (MRSA), its specific mechanism in MRSA is worthy of investigation, which is the focus of this study. Initially, the collected S. aureus strains were treated with luteolin. Then, the minimum inhibitory concentration (MIC) of luteolin against the S. aureus strains was measured by the broth microdilution. The growth curves, biofilm formation, and cytotoxicity of treated S. aureus were detected using a microplate reader. The live and dead bacteria were evaluated using confocal laser scanning microscopy, the bacterial morphology was observed using scanning electron microscopy, and the S. aureus colony-forming unit (CFU) numbers were assessed. The levels of alpha hemolysin (α-hemolysin), delta hemolysin (δ-hemolysin), and hlaA were detected via western blot and RT-PCR. The mortality of mouse model with S. aureus systemic infection was analyzed, and the levels of IL-6, IL-8, IL-10, and TNF-α were quantitated using ELISA. Concretely, the MIC of luteolin against MRSA N315 was 64 µg/mL. Luteolin at 16 µg/mL did not affect the growth of MRSA N315, but inhibited the biofilm formation and CFU, and promoted the morphological changes and death of MRSA N315. Luteolin decreased the cytotoxicity and the levels of α-hemolysin, δ-hemolysin, and hlaA in MRSA N315, elevated MRSA-reduced mice survival rate, and differentially modulated the inflammatory cytokine levels in MRSA-infected mice. Collectively, luteolin inhibits biofilm formation and cytotoxicity of MRSA via blocking the bacterial toxin synthesis.

A network pharmacology approach to investigate the anticancer mechanism of cinobufagin against hepatocellular carcinoma via downregulation of EGFR-CDK2 signaling.

Hepatocellular carcinoma (HCC) is one of the deadliest cancers with high mortality and poor prognosis, and the investigation on new approaches and effective drugs for HCC therapy is of great significance. In our study, we demonstrate that treatment with cinobufagin, a natural compound isolated from traditional chinese medicine Chansu, reduces proliferation and the colony formation capacity of the human hepatoma cells in vitro, in addition, cinobufagin induces mitotic arrest in human hepatoma cells. The results of a network pharmacology-based analysis show that EGFR, MAPK1, PTK2, CDK2, MAPK3, ESR1, CDK1, PRKCA, AR, and CSNK2A1 are the key targets involved in the anti-tumor activities of cinobufagin, additionally, several signaling pathways such as proteoglycans in cancer, pathways in cancer, HIF-1 signaling pathway, VEGF signaling pathway, ErbB signaling pathway, and PI3K-AKT signaling pathway are identified as the potential pathways involved in the inhibitory effects of cinobufagin against HCC. Furthermore, at the molecular level, we find that cinobufagin decreases EGFR expression and CDK2 activity in human hepatoma cells. Inhibition of EGFR or CDK2 expression could not only suppress the growth of tumor cells but also enhance the inhibitory effects of cinobufagin on the proliferative potential of human hepatoma cells. We also demonstrate that EGFR positively regulates CDK2 expression. Furthermore, EGFR inhibitor gefitinib or CDK2 inhibitor CVT-313 synergistically enhances anticancer effects of cinobufagin in human hepatoma cells. Taken together, these findings indicate that cinobufagin may exert antitumor effects by suppressing EGFR-CDK2 signaling, and our study suggests that cinobufagin may be a novel, promising anticancer agent for the treatment of HCC.

Disruption of YY1-EZH2 Interaction Using Synthetic Peptides Inhibits Breast Cancer Development.

Enhancer of zeste homolog 2 (EZH2) is a methyltransferase to mediate lysine 27 trimethylation in histone H3 (i.e., H3K27me3) and repress gene expression. In solid tumors, EZH2 promotes oncogenesis and is considered a therapeutic target. As a transcription factor, Yin Yang 1 (YY1) recruits EZH2 through its oncoprotein binding (OPB) domain to establish gene repression. In this study, we mapped the YY1 protein binding (YPB) domain on EZH2 to a region of 27 amino acids. Both YPB and OPB domain synthetic peptides could disrupt YY1EZH2 interaction, markedly reduce breast cancer cell viability, and efficiently inhibit tumor growth in a xenograft mouse model. We analyzed MDA-MB-231 cells treated with YPB, OPB, and control peptides by chromatin immunoprecipitation DNA sequencing (ChIP-seq) using an antibody against H3K27me3. YPB and OPB treatments altered H3K27me3 on 465 and 1137 genes, respectively, compared to the control. Of these genes, 145 overlapped between the two peptides. Among them, PTENP1, the PTEN pseudogene, showed reduced H3K27me3 signal when treated by either YPB or OPB peptide. Consistently, the two peptides enhanced both PTENP1 and PTEN expression with concomitantly reduced AKT activation. Further studies validated PTENP1's contribution to the anticancer activity of YPB and OPB peptides.

Berberine attenuates hepatic steatosis and enhances energy expenditure in mice by inducing autophagy and fibroblast growth factor 21.

BACKGROUND AND PURPOSE: Berberine, a compound from rhizome coptidis, is traditionally used to treat gastrointestinal infections, such as bacterial diarrhoea. Recently, berberine was shown to have hypoglycaemic and hypolipidaemic effects. We investigated the mechanisms by which berberine regulates hepatic lipid metabolism and energy expenditure in mice. EXPERIMENTAL APPROACH: Liver-specific SIRT1 knockout mice and their wild-type littermates were fed a high-fat, high-sucrose (HFHS) diet and treated with berberine by i.p. injection for five weeks. Mouse primary hepatocytes and human HepG2 cells were treated with berberine and then subjected to immunoblotting analysis and Oil Red O staining. KEY RESULTS: Berberine attenuated hepatic steatosis and controlled energy balance in mice by inducing autophagy and FGF21. These beneficial effects of berberine on autophagy and hepatic steatosis were abolished by a deficiency of the nutrient sensor SIRT1 in the liver of HFHS diet-fed obese mice and in mouse primary hepatocytes. SIRT1 is essential for berberine to potentiate autophagy and inhibit lipid storage in mouse livers in response to fasting. Mechanistically, the berberine stimulates SIRT1 deacetylation activity and induces autophagy in an autophagy protein 5-dependent manner. Moreover, the administration of berberine was shown to promote hepatic gene expression and circulating levels of FGF21 and ketone bodies in mice in a SIRT1-dependent manner. CONCLUSIONS AND IMPLICATIONS: Berberine acts in the liver to regulate lipid utilization and maintain whole-body energy metabolism by mediating autophagy and FGF21 activation. Hence, it has therapeutic potential for treating metabolic defects under nutritional overload, such as fatty liver diseases, type 2 diabetes and obesity.

Clinical effectiveness of Novolin® 30R versus Lantus® combined with Glucobay® treatment in elderly patients with type 2 diabetes mellitus controlled by oral hypoglycaemic agents: A randomized study.

OBJECTIVE: To compare the treatment effects of Novolin® 30R(a), versus Lantus(®a) combined with acarbose (Glucobay®), in elderly patients with type 2 diabetes mellitus. METHODS: Patients (aged > 60 years) with type 2 diabetes mellitus were randomized to receive either Novolin® 30R(a) (initial dose 0.5 IU/kg) or Lantus(®a) (initial dose 0.2 IU/kg) combined with 50 mg acarbose. After a 32-week treatment period, the following parameters were measured: blood glucose control; blood lipid levels; body mass index; proportion of patients achieving a glycosylated haemoglobin (HbA1c) level <7.5%; rate of hypoglycaemic events; change in fasting blood glucose levels from baseline in patients stratified according to their baseline HbA1c level. RESULTS: A total of 188 patients were enrolled in the study. After 32 weeks' treatment, compared with baseline levels, there were significant reductions in FBG, 2 h-postprandial blood glucose during an oral glucose tolerance test, HbA1c, total cholesterol, triglycerides and low-density lipoprotein cholesterol values in both groups. Although there were fewer hypoglycaemic events in the Lantus® combined with Glucobay® group compared with the Novolin® 30R group, the difference was not significant. CONCLUSION: Novolin® 30R and Lantus® combined with acarbose both had beneficial effects on blood glucose control and blood lipid levels in elderly patients with type 2 diabetes mellitus.