Protective Effect of Schisandrin on CORT-Induced PC12 Depression Cell Model by Inhibiting Cell Apoptosis In Vitro.

Background: In recent years, the incidence of depression is on the rise. Our paper proposed to study the protective effects of Schisandrin on CORT-induced PC12 depressive cell model and the underlying mechanisms. Methods: The in vitro models of PC12 were established using corticosterone (CORT). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was used to screen the effective concentration of Schisandrin, and the models of PC12 were treated with low, medium, and high concentrations of Schisandrin. The cell activity of each group was detected by MTT assay. The LDH activity in each group of cells was detected by lactate dehydrogenase (LDH) kit. Apoptosis rate of each group was detected by Annexin V-FITC apoptosis assay kit. Mitochondrial membrane potential of each group of cells was detected by mitochondrial membrane potential kit. The protein expression levels of Caspase-3, Bax, and Bcl-2 in each group of cells were detected by western blot. Results: The treatment of Schisandrin significantly increased the cell viability in models of PC12. In addition, the results of LDH activity suggested that Schisandrin significantly reduced LDH content in models of PC12. Consistently, Schisandrin reduced the mitochondrial membrane potential of CORT-induced PC12 depressive cell model. Furthermore, Schisandrin effectively reduced the number of apoptotic cells and inhibited the expression of proapoptotic-related proteins (cleaved Caspase-3 and Bax) but increased the antiapoptotic-related protein (Bcl-2) in the models of PC12. Conclusions: Protective effects of Schisandrin on CORT-induced PC12 depressive cell model by inhibiting cells apoptosis in vitro.

Protective effect of isopsoralen on UVB-induced injury in HaCaT cells via the ER and p38MAPK signaling pathways.

This study investigated the protective effect of isopsoralen on UVB-induced damage in HaCaT cells and its molecular mechanism. The cytotoxicity of isopsoralen and its effects on the viability of HaCaT cells were examined using the MTT assay. The effects of UVB irradiation and isopsoralen on the intracellular glutathione (GSH-PX), superoxide dismutase (SOD), malondialdehyde (MDA), and reactive oxygen species (ROS) content were examined using commercially available assay kits. Further, the effects of UVB irradiation and isopsoralen on the levels of the inflammatory cytokines TNF-α, IL-6, and IL-1α were examined using enzyme-linked immunosorbent assay. Finally, we examined the effect of adding the estrogen receptor (ER) antagonist ICI182780,780 and the p38MAPK antagonist SB203580 on the changes in inflammatory cytokines induced by isopsoralen treatment and UVB irradiation. Isopsoralen pretreatment markedly inhibited UVB-induced reduction in the viability and proliferation of HaCaT cells. Isopsoralen also reduced UVB-induced increase in the expression of the inflammatory cytokines and the level of free radicals (ROS and MDA), and reversed the UVB-induced suppression of antioxidant activity. Additionally, inhibition of ER and p38MAPK via the addition of their respective antagonists reversed the observed anti-inflammatory effects of Isopsoralen. Isopsoralen can efficiently provide protection against UVB-induced damage in HaCaT cells brought about via oxidation and inflammatory reactions, and the underlying mechanisms involve the ER and p38MAPK pathways. Therefore, Isopsoralen could be used in therapeutic solutions for UVB-induced skin conditions. PRACTICAL APPLICATIONS: Isopsoralen shows antioxidant and anti-inflammatory effects. As natural, healthy, and effective additives, isopsoralen has been widely used in cosmetics and botanical medicine products. The results of this study reveal the molecular mechanisms underlying isopsoralen effects, showing that isopsoralen reverses the effects of UVB irradiation regulating ER and p38MAPK signaling pathways. Consequently, isopsoralen regulates the expression of ER and p38MAPK signaling pathways, thereby reducing the activation of antioxidant and anti-inflammatory activity. These findings suggest that isopsoralen can be used as the base ingredient for antiphotoaging cosmetics and botanical medicine products. This study provides both theoretical and experimental background for isopsoralen deep processing and utilization.

Tissue Distribution of Total Flavonoids Extracts of Drynariae Rhizoma in Young and Old Rats by UPLC-MS/MS Determination.

Drynariae Rhizoma (Kunze ex Mett.) J. Sm. has been extensively used in China, Japan, and Korea to treat osteoporosis and tonify kidneys. A rapid and validated UPLC-MS/MS method for simultaneous determination of the seven analytes including neoeriocitrin, luteolin-7-O-ß-D-glucoside, astragalin, naringin, eriodictyol, naringenin, and kaempferol in rats' various tissues (heart, liver, spleen, lung, kidney, stomach, brain, uterus, ovary, and small intestine) using quercetin as the internal standard (IS) was developed after oral administration of TFDR to rats. Tissues samples were retreated by protein precipitation with methanol. The chromatographic separation was performed using an ACQUITY UPLC™ BEH C18 column (2.1 × 50 mm; 1.7 µm) at 35°C. The mobile phase consisted of 1% acetic acid in water as the aqueous phase (A) and 100% acetonitrile as the organic phase (B). All analytes and IS were quantified through electrospray ionization in positive ion multiple reaction monitoring (MRM) mode. MS transitions were m/z 597.5 ⟶ 289.2 for neoeriocitrin, m/z 449.1 ⟶ 287.1 for luteolin-7-O-ß-D-glucoside, m/z 449.1 ⟶ 287.1 for astragalin, m/z 581.5 ⟶ 273.2 for naringin, m/z 289.2 ⟶ 153.1 for eriodictyol, m/z 273.2 ⟶ 153.1 for naringenin, m/z 287.1 ⟶ 153.1 for kaempferol, and m/z 303.2 ⟶ 153.1 for quercetin (IS). The mean extraction recovery of the seven analytes and IS in tissue samples at three levels of quality control (QC) samples ranged from 82.72% to 118.57%, and the RSD was ≤14.98%. The intraday and interday precision (RSD) were all less than 14.98%, and the accuracy (RE) ranged from -13.96% to 14.96%, which indicated that the present method was not an issue. Tissues distribution showed neoeriocitrin, luteolin-7-O-ß-D-glucoside, astragalin, naringin, and naringenin could transfer across the blood-brain barrier, which may form the basis of TFDR entering the brain to play an anti-AD role. Compared with the 8-month-old rats, a higher concentration of naringin was found in the ovaries of the 18-month-old rats; this result indicated that it may regulate the autonomic nervous dysfunction of the cerebrospinal system through the hypothalamus-pituitary-ovary axis, thus playing an anti-AD role, but further research is needed. Naringenin, eriodictyol, and kaempferol have a higher concentration in the liver and kidney; this phenomenon may be related to the traditional Chinese medicine theory that there is a definite relationship between the liver and kidney meridian. These results provide reliable data support for further study of the pharmacological mechanism of TFDR, formulation of drug delivery schemes, and development of new Chinese medicines in the treatment of AD.

Changes in liver function of Tibetans with different BMI during the early stage of migration to plain / 中国学校卫生

Objective@#To explore the changes in liver function of Tibetan youth living in plateau with different body mass index (BMI) during the early stage of migration to the plain, and to provide scientific basis for high attitude de adaptation.@*Methods@#A total of 3 035 Tibetan youth who firstly migrated to the plain (Shaanxi) from the plateau (Tibet) were selected as the research subjects, and were screened for symptoms of plateau de adaptation. Participants were divided into four groups: underweight, normal weight, overweight and obese, and received liver function test on the 3rd, 6th, 9th dayafter migration, respectively. Chi square test was used to detect the abnormal rate of liver function indicators among each group, and binary Logistic regression analysis was used to explore the relationship between BMI and abnormal liver function indicators.@*Results@#The alanine aminotransferase(ALT), Aspartate aminotransferase(AST), γ glutamyl transpeptidase (GGT) of overweight Tibetan male and obese Tibetan male and female adolescents, the total bile acid (TBA) of overweight Tibetan male and obese Tibetan female were higher than those of the normal weight group at the early stage of de adaptation( P <0.05). With the de adaptation for 3, 6, 9 days, the indexes showed an overall upward trend, including: direct bilirubin (DBIL) in overweight male and female adolescents, total protein (TP) and globulose (GLOB) in obese female adolescents( P <0.05). The abnormal rate of overweight group (male ALT: 13.9%), obesity group (male and female ALT, GGT: 34.3%, 26.7%, 11.4%, 13.3%; female AST:10.0%) was significantly higher than that in underweight (2.8%, 3.5%, 0, 1.0%, 1.5%) and normal group(3.5%, 3.4%, 0.9%, 3.6%, 4.1%)( χ 2=48.07, 20.55, 20.55, 17.93, 10.23 , P <0.05). Binary Logistic regression analysis showed that after adjusting for age and gender, overweight was positively correlated with abnormal ALT( OR=2.10, 95%CI =1.20-3.62). Obesity was positively correlated with abnormal ALT( OR=5.50, 95%CI =4.23-7.40) and GGT( OR=4.10, 95%CI =2.03-6.74)( P <0.05).@*Conclusion@#During the early stage of migration to the plain among Tibetan youth living on the plateau, changes in liver function indicators are related to BMI. Overweight and obese Tibetans have a higher abnormal rate of liver damage indicators. It is suggested that individuals with high risk of obesity should undergo health examination and medical supervision when migrates from plateau to plain.

Direct electrochemistry of silver nanoparticles-decorated metal-organic frameworks for telomerase activity sensing via allosteric activation of an aptamer hairpin.

A direct electrochemistry of silver nanoparticles (AgNPs)-anchored metal-organic frameworks (MOFs) is developed for detection of telomerase activity based on allosteric activation of an aptamer hairpin. AgNPs in situ decorated on PCN-224 (AgNPs/PCN-224) constituted the direct electrochemical labels that were further biofunctionalized by recognition moiety of streptavidin (SA). To achieve the target biosensing, an allosteric hairpin-structured DNA was elaborately designed for signal transduction. The presence of telomerase elongated its primer in the hairpin to displace partial stem strand, thus resulted in the formation of SA aptamer-open structure. Through the specific interaction with aptamer, SA-biofunctionalized AgNPs/PCN-224 probe was attached onto the electrode surface, generating electrochemical signal at + 0.072 V of AgNPs centralized by MOF structure. The direct electrochemical biosensor showed target activity-dependent response from 1.0 × 10-7 to 1.0 × 10-1 IU L-1 with a detection limit of 5.4 × 10-8 IU L-1. Moreover, the sensor was applied in evaluation of telomerase activity in living cancer cells. The established electrochemical detection approach in this work avoids the critical deoxygenation conditions and additional electrocatalytic reagents, which opens a novel biosensing perspective for direct electrochemistry of MOF-based nanocomposites.

Protective Role Of Naringenin Against Aß25-35-Caused Damage via ER and PI3K/Akt-Mediated Pathways.

Senile plaque accumulation and neurofibrillary tangles are primary characteristics of Alzheimer's disease. We aimed to assess the protective functions of naringenin against ß-amyloid protein fragment 25-35 (Aß25-35)-caused nerve damage in differentiated PC12 cells, and study the potential mechanisms. We evaluated cell viability and apoptosis using the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) test and flow cytometry, respectively. Moreover, we measured protein kinase B (Akt), glycogen synthase kinase-3ß (GSK-3ß), and caspase-3 activity via western blotting and RT-PCR. We found that naringenin protected cell against Aß25-35-caused nerve damage by increasing cell viability, promoting Akt and GSK3ß activation, and inhibiting cell apoptosis and caspase-3 activity. However, treatment with the estrogen receptor (ER) antagonist ICI182, 780 or phosphatidylinositol-3-kinase (PI3K) inhibitor LY294002 suppressed the effects of naringenin. Our results suggested that naringenin could effectively suppress Aß25-35-caused nerve damage in PC12 cells by regulating the ER and PI3K/Akt pathways.

Effects of wogonin on the mechanism of melanin synthesis in A375 cells.

The present study aimed to investigate the effect of wogonin on the mechanism of melanin synthesis in the A375 melanoma cell line. A375 cells, cultured in vitro, were treated with wogonin and the activity of tyrosinase (TYR) and melanin synthesis were examined via MTT assay, L-dopa oxidation assay and an NaOH lysis assay. Protein expression levels of TYR and c-Jun N-terminal kinase (JNK) were examined via western blotting. mRNA expression levels of TYR, tyrosinase related protein (TRP)-1, TRP-2, extracellular signal-regulated kinase (ERK)-1, ERK-2 and JNK-2 were analyzed by reverse transcription-quantitative polymerase chain reaction. Furthermore, the effect of wogonin on estrogen receptor inhibitor (ICI182780) and ERK pathway inhibitor (U0126) was investigated. Safe doses of wogonin (10, 1, 10-1, 10-2 or 10-3 µmol/l) significantly inhibited melanin synthesis and TYR activity (P<0.05). Wogonin (10 µmol/l) inhibited the protein expression levels of TYR, JNK and mRNA expression levels of TYR, TRP-1, TRP-2, ERK-1, ERK-2, JNK-2 in A375 cells (P<0.01). The estrogen receptor inhibitor, ICI182780, and MEK inhibitor, U0126, significantly reversed the effects of wogonin on protein and mRNA expression levels of TYR, TRP-1, TRP-2, ERK-1, ERK-2 and JNK-2 (all P<0.01). To conclude, the present study identified that wogonin is able to inhibit the synthesis of melanin in A375 cells, through inhibiting protein and mRNA expression levels of TYR, TRP-1, and TRP-2, and ERK1, ERK2 and JNK2, respectively.

Preliminary study of urine metabolism in type two diabetic patients based on GC-MS.

OBJECTIVE: Comparative study of type 2 diabetes and healthy controls by metabolomics methods to explore the pathogenesis of Type II diabetes. METHODS: Gas chromatography - mass spectrometry (GC-MS) with a variety of multivariate statistical analysis methods to the healthy control group 58 cases, 68 cases of Type II diabetes group were analyzed. Chromatographic conditions: DB-5MS column; the carrier gas He; flow rate of 1 mL·min(-1), the injection volume 1 uL; split ratio is 100: 1. MS conditions: electron impact (EI) ion source, an auxiliary temperature of 280°C, the ion source 230°C, quadrupole 150°C; mass scan range 30~600 mAu. RESULTS: Established analytical method based on urine metabolomics GC-MS of Type II diabetes, determine the urine succinic acid, L-leucine, L-isoleucine, tyrosine, slanine, acetoace acid, mannose, L-isoleucine, L-threonine, Phenylalanine, fructose, D-glucose, palmi acid, oleic acid and arachidonic acid were significantly were significantly changed. CONCLUSION: Based on metabolomics of GC-MS detection and analysis metabolites can be found differences between type 2 diabetes and healthy control group, PCA diagram can effectively distinguish Type II diabetes and healthy control group, with load diagrams and PLS-DA VIP value metabolite screening, the resulting differences in metabolic pathways involved metabolites, including amino acid metabolism, lipid metabolism, glucose metabolism and energy metabolism.

Effects of bavachin and its regulation of melanin synthesis in A375 cells.

The aim of the present study was to investigate the effect of bavachin treatment on A375 cells and the regulation of melanin synthesis. The cultured A375 cells in vitro were treated with bavachin; and the effect of bavachin on cell activity, tyrosinase (TYR) activity and melanin synthesis were respectively tested by the MTT assay, L-dopa oxidation assay and the NaOH lysis assay. The expression levels of TYR and c-Jun N-terminal kinases (JNK) proteins were tested by western blot analysis. The expression levels of TYR, tyrosinase-related protein-1 (TRP-1), TRP-2, extracellular signal-regulated kinase 1 (ERK1), ERK2 and JNK2 mRNA were tested by the reverse transcription-polymerase chain reaction assay. Simultaneously, the effect of estrogen receptor inhibitor (ICI182780) and ERK pathway inhibitor (U0126) was also tested on A375 cells following bavachin. The safe dose of bavachin significantly inhibited melanin synthesis and TYR activity. Bavachin (10 µmol/l) inhibited the expression of TYR and JNK proteins, and the expression of TYR, TRP-1, TRP-2, ERK1, ERK2 and JNK2 mRNA in A375 cells. ICI182780 and U0126 could significantly reverse the bavachin treatment on the protein expression levels and the mRNA expression of TYR, TRP-1, TRP-2, ERK1, ERK2 and JNK2. In conclusion, bavachin inhibited the synthesis of melanin on A375 cells by inhibiting the protein and mRNA expression of TYR, TRP-1, TRP-2, ERK1, ERK2 and JNK2.

Determination of the secondary structure of proteins in different environments by FTIR-ATR spectroscopy and PLS regression.

The secondary structures of proteins (alpha-helical, beta-sheet, beta-turn, and random coil) in the solid state and when bound to polymer beads, containing immobilized phenyl and butyl ligands such as those as commonly employed in hydrophobic interaction chromatography, have been investigated using FTIR-ATR spectroscopy and partial least squares (PLS) methods. Proteins with known structural features were used as models, including 12 proteins in the solid state and 7 proteins adsorbed onto the hydrophobic surfaces. A strong PLS correlation was achieved between predictions derived from the experimental data for 4 proteins adsorbed onto the phenyl-modified beads and reference data obtained from the X-ray crystallographic structures with r(2) values of 0.9974, 0.9864, 0.9924, and 0.9743 for alpha-helical, beta-sheet, beta-turn, and random coiled structures, respectively. On the other hand, proteins adsorbed onto the butyl sorbent underwent greater secondary structural changes compared to the phenyl sorbent as evidenced from the poorer PLS r(2) values (r(2) are 0.9658, 0.9106, 0.9571, and 0.9340). The results thus indicate that the secondary structures for these proteins were more affected by the butyl sorbent, whereas the secondary structure remains relatively unchanged for the proteins adsorbed onto the phenyl sorbent. This study has important ramifications for understanding the nature of protein secondary structural changes following adsorption onto hydrophobic sorbent surfaces. This knowledge could also enable the development of useful protocols for enhancing the chromatographic purification of proteins in their native bioactive states.