Effect of hydroxysafflor yellow A on depressive-like behavior and expression of GABAAR protein in hippocampus of chronic restraint stress model mice / 中华行为医学与脑科学杂志

Objective:To investigate the effects of hydroxysafflor yellow A (HSYA) on depressive-like behavior and expression of type A γ-aminobutyric acid receptor(GABAAR)in hippocampus of chronic restraint stress model mice.Methods:The SPF grade male C57BL/6C mice were divided into Control group, HSYA group, Model group, Model + HSYA group and Model + fluoxetine group according to random number table method, with 12 mice in each group.Mice model of depression was established by chronic restraint stress.Mice in HSYA group and Model+ HSYA group were intraperitoneally injected with HSYA(20 mg/kg), mice in Model+ fluoxetine group were injected intraperitoneally with fluoxetine (10 mg/kg), and mice in Control group and Model group administered with 0.9% sodium chloride solution intraperitoneally once a day for 14 days.Then, the forced swimming test (FST) and tail suspension test (TST) were performed to evaluate the depressive-like behavior of mice, and the protein expression levels of different subtypes of GABAAR in the hippocampus of mice were determined by Western blot.SPSS 19.0 and GraphPad Prism 8.0 software were used for data statistical analysis and mapping.One-way ANOVA was used for comparison among groups, and Tukey-HSD test was used for further pairwise comparison.Results:(1) In the behavioral tests, there were significant differences in swimming immobility time of FST and tail suspension immobility time of TST among the five groups ( F=21.59, 20.81, both P<0.05). The swimming immobility time ((143.91±9.97) s) and tail suspension immobility time (( 107.00±6.54) s) in Model group were higher than those in Control group ((52.92±6.70) s, ( 43.50±5.96) s, both P<0.05). There were no significant difference in swimming immobility time and tail suspension immobility time between Model+ HSYA group ((26.17±7.69)s, ( 20.17±7.89)s) and Model+ fluoxetine group ((61.60±16.22)s, (34.14±10.74)s)(both P>0.05), but the swimming immobility time and tail suspension immobility time in these two groups were lower than those in Model group (both P<0.05). (2) The Western blot results showed that there were significant differences in the expression of GABAARβ1 and GABAARβ2 protein in hippocampus among the four groups ( F=12.21, 11.40, both P<0.05). The expression levels of GABAARβ1(45.60±10.76) and GABAARβ2 (46.27±4.82) protein in hippocampus of Model group were lower than those in Control group ((100.00±3.44), (100.00±3.26), both P<0.05). Compared to Model group, the expression of GABAARβ1 (79.91±5.00) and GABAARβ2 (79.08±5.53) protein in hippocampus of Model+ HSYA group were higher (both P<0.05). In addition, the expression of GABAARα1 and GABAARγ1 proteins in hippocampus were not significantly different among the four groups( F=0.23, 0.10, both P>0.05). Conclusion:HSYA can effectively alleviate depressive-like behavior in depression model mice, which may be related with the upregulation of GABAARβ1 and GABAARβ2 of hippocampus tissue.

Establishment of HPLC Fingerprint of Coptis chinensis Inflorescence and Its Spectrum-effect Relationship with Antioxidant and Antibacterial Effects / 中国药房

OBJECTIVE：To establish HPLC fingerprint of Coptis chinensis inflorescence，and study its spectrum-effect relationship with antioxidant and antibacterial effects. METHODS ：Taking 14 batches of C. chinensis inflorescence from different producing areas as the object ，HPLC method was adopted. The determination was performed on Supersil C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid solution（gradient elution ）at the flow rate of 1.0 mL/min. The column temperature was set at 25 ℃. The detection wavelength was set at 329 nm，and sample size was 10 μL. The fingerprints of 14 batches of C. chinensis inflorescence were established by Similarity Evaluation System of TCM Fingerprint （2012 A edition ），and the similarity evaluation and common peak identification were carried out. Taking DPPH free radical scavenging rate and hydroxyl radical scavenging rate as antioxidant effects index ，relative antibacterial activity （Escherichia coli ）as antibacterial effect index ， SPSS 21.0 software was adopted to analyze the Pearson correlation between common peaks of C. chinensis inflorescence and above efficacy indexes ；their spectrum-effect relationship was established and validated. RESULTS ：A total of 7 common peaks were obtained in HPLC fingerprint of C. chinensis inflorescence，and the similarity was no less than 0.916. No. 5 peak was identified as berberine hydrochloride. Seven common peaks were positively correlated with DPPH free radical scavenging rate ；No. 1-3，4，6，7 peaks were positively correlated with hydroxyl radical scavenging rate ，while No. 5 peak was negatively correlated with hydroxyl radical scavenging rate. There was a positive correlation between No. 5 peak and antibacterial activity in vitro . After validation ， relative error between the predicted values and the measured values of DPPH free radical scavenging rate ，hydroxyl radical scavenging rate and relative antibacterial activity was 0.92％- 14.5％ . CONLUSIONS ：The established spectrum-effect relationship model can be used to evaluate antioxidant andantibacterial effects of C. chinensis inflorescence. The chemical components represented by No. 1，2，3，4，6，7 peaks are the material basis of antioxidant effect of C. chinensis inflorescence， and berberine hydrochloride is the material basis of antibacterial effect.

Simultaneous genotyping of DRB1/3/4/5 loci by oligonucleotide microarray.

Matching of the HLA antigens for donor-recipient in transplantation, disease predisposition or protection, population studies, and forensic testing requires accurate but simple typing methods. Here, we describe a DNA-based tissue-typing assay that determines the haplotype of the DRB1/3/4/5 loci in hy-bridization of oligonucleotide array after sample amplification. Using this multianalyte DNA hybridization system, we analyzed seven regions of exon 2 of DRB loci that have single-base discrimination. Thirty-six oligonucleotide probes complementary to the alleles of interest were immobilized on each microslide. The efficiency and specificity of identifying DRB genotypes using the oligonucleotide arrays was evaluated by blinded analysis of 147 samples from reference standards and subjects. The established method provides a rapid and inexpensive DRB "low-resolution" typing tool for prescreening a large number of samples.

Estimation of relative allele frequencies of single-nucleotide polymorphisms in different populations by microarray hybridization of pooled DNA.

Single-nucleotide polymorphisms (SNPs) are considered useful polymorphic markers for genetic studies of polygenic traits. A new practical approach to high-throughput genotyping of SNPs in a large number of individuals is needed in association study and other studies on relationships between genes and diseases. We have developed an accurate and high-throughput method for determining the allele frequencies by pooling the DNA samples and applying a DNA microarray hybridization analysis. In this method, the combination of the microarray, DNA pooling, probe pair hybridization, and fluorescent ratio analysis solves the dual problems of parallel multiple sample analysis, and parallel multiplex SNP genotyping for association study. Multiple DNA samples are immobilized on a slide and a single hybridization is performed with a pool of allele-specific oligonucleotide probes. The results of this study show that hybridization of microarray from pooled DNA samples can accurately obtain estimates of absolute allele frequencies in a sample pool. This method can also be used to identify differences in allele frequencies in distinct populations. It is amenable to automation and is suitable for immediate utilization for high-throughput genotyping of SNP.