ABSTRACT
The present study established the spectrum-effect relationship model of flavonoids in Citri Reticulatae Pericarpium(CRP) from 15 batches of Liujunzi Decoction and statistically analyzed the correlation between chemical peaks and efficacy to identify the main effective components. HPLC fingerprints of flavonoids in CRP from 15 batches of Liujunzi Decoction were established. HPLC analysis was carried out on the Venusil XBP C_(18)(L) column(4.6 mm×250 mm, 5 μm) at 30 ℃ with acetonitrile-water(containing 0.1% formic acid) as mobile phase for gradient elution, a flow rate of 1.0 mL·min~(-1), and detection wavelength of 300 nm to obtain chemical fingerprints. Additionally, the effects of flavonoids from CRP in 15 batches of Liujunzi Decoction on the content of GAS, MTL, and VIP, TFF3 mRNA expression, and percentage of CD3~+ T-cells of model rats with spleen deficiency were determined. The spectrum-effect relationship model was established by gray correlation analysis. The results showed that the main characteristic peaks with great contribution to the regulation of gastrointestinal tract were peak 16(vicenin-2), peak 63(sinensetin), peak 64(isosinensetin), peak 65(nobiletin), peak 67(3,5,6,7,8,3',4'-heptemthoxyflavone), peak 68(tangeretin), and peak 69(5-desmethylnobiletin). Therefore, there was a linear correlation between flavonoids from CRP in Liujunzi Decoction and the efficacy, and the medicinal effect was achieved by multi-component action. This study is expected to provide a new idea for exploring the material basis of the effect, i.e., regulating qi prior to replenishing qi, of CRP in Liujunzi Decoction.
Subject(s)
Animals , Rats , Citrus/chemistry , Drugs, Chinese Herbal , Flavonoids/pharmacology , Hormones , RNA, Messenger/genetics , SpleenABSTRACT
Zixue was first recorded in detail in Qianjin Yifang, and practitioners of later generations renamed it as Zixuesan and Zixuedan, which is the representative prescription of antipyretic preparations and has high clinical application and research value. At present, there have been many studies on the pharmacological effect and clinical application of Zixue, but the medical books of the past dynasties have slightly different records on Zixue, which has brought some difficulties to the reasonable clinical application and research of this prescription. In addition to the introduction of Zixuesan, which is different from the ancient recipe, the researchers have questions about which development method can reasonably inherit and develop the classical preparation. The authors intend to summarize the historical evolution of "formula" and "preparation" of Zixue, in order to clarify its historical context, which can provide a reference for the modern research and development of this formula, and provide a new ideas for the in-depth exploration of other classic preparations.
ABSTRACT
<p><b>OBJECTIVE</b>To construct Epithelia Membrane Protein 1 gene-deficient in human fetal nucleus pulposus model by lentivirus-mediated RNA interference for building a platform for illustrating the biomechanisms role of EMP-1 during human intervertebral disc degeneration.</p><p><b>METHODS</b>The lentivirus vector with shRNA targeting EMP-1 mRNA was transected into 293FT cells by liposome. Then the lentivirus supernatant was obtained and used for infecting human fetal nucleus pulposus. The expression of GFP was observed under fluorescence microscope after 48 h. The viral particles were collected at 72 h after transfection. The efficacy of gene interference was tested by Western blot and Real-time RT-PCR. Analysis the results of the fluorescent microscope scenes and get the average values of EMP-1/GAPDH by detected the interference efficiency of various interference DNA sequences with western blot and semi quantitative RT-PCR methods.</p><p><b>RESULTS</b>The lentivirns with high titer were obtained and the EMP-1 gene deficient cell strains were obtained. Semi quantitative RT-PCR and Western blot proved the average values of EMP-1/GAPDH decreased from 0.46 to 0.32 and 0.5 to 0.25 (P < 0.01).</p><p><b>CONCLUSION</b>Lentivirus packaging technology can be mastered skillfully. EMP-1 gene-deficient cell models are successfully established.</p>