ABSTRACT
ObjectiveTo explore the pharmacodynamic ingredients of Zhenqi Fuzheng granules （ZFG） for immunomodulatory through spectrum-effect relationship analysis， which provides experimental basis for improving the quality standard of ZFG. MethodEighteen batches of ZFG from six manufacturers were collected for analysis. The fingerprints were established by high performance liquid chromatography （HPLC）. Acetonitrile （A）-0.1% formic acid aqueous solution （B） were adopted as the mobile phase with gradient elution （0-15 min， 5%A； 15-23 min， 5%-8%A； 23-30 min， 8%-11%A； 30-45 min， 11%-18%A； 45-60 min， 18%-21%A； 60-67 min， 21%-23%A； 67-90 min， 23%-37%A）， the detection wavelength was 220 nm. Chemometric analysis such as similarity analysis and hierarchical cluster analysis （HCA） were subsequently used to analyze the similarities and chemical differences among these samples. A cyclophosphamide-induced immunodeficiency mouse model was used to evaluate the immune-enhancing effects of the products from different manufacturers. The spectrum-effect relationship between HPLC fingerprints and the immunomodulatory effects was examined using Spearman bivariate correlation analysis. HPLC coupled with mass spectrometry （HPLC-MSn） was used to identify the spectrum-effect related peaks with electrospray ionization， positive and negative ion modes， and scanning range of m/z 100-1 500. ResultThe HPLC fingerprint of ZFG was established， and twenty peaks with good resolution were selected as common peaks. The results of quality analysis and pharmacodynamic test showed there were significant differences in both ingredients content and immune-enhancing effects of ZFG from different manufacturers. Through spectrum-effect relationship study， twelve peaks were screened as bioactive ingredients peaks. Thereafter， eight peaks among them were subsequently identified by HPLC-MSn. They were salidroside （peak 2）， echinacoside （peak 5）， calycosin-7-glucoside （peak 6）， isomer of specnuezhenide （peak 7）， isonuezhenide （peak 9）， calycosin （peak 11）， nuezhenide G13 or oleonuezhenide （peak 14）， and formononetin （peak 18）， respectively. ConclusionThere are differences in quality and efficacy of ZFG produced by different manufacturers. Through spectrum-effect relationship analysis， the medicinal ingredients of ZFG for immune-enhancing effects are screened， which can provide reference for the improvement of its quality standard.
ABSTRACT
Rat models of liver fibrosis were made by carbon tetrachloride, and the serum levels of AST, ALT, γ-GT, MDA, GSH-px, SOD were detected, serum markers of PCⅢ, IV-C, LN, HA were detected by ELISA method. HE and Masson staining were conducted in hepatic tissues to observe pathological variations. Collagen Ⅲ, TGF-β, α-SMA, E-cadherin were detected by Western blot. The curative effect of the extract of Ornithogalum caudatum on rat liver fibrosis induced by CCl4was observed and the mechanism was discussed. The experiment results showed that the extract of O. caudatum (50, 150, 500 mg•kg⁻¹) obviously decreased the serum levels of AST, ALT, γ-GT, MDA, increased the serum levels of GSH-px, SOD, decreased the expression of serum markers of PCⅢ, IV-C, LN, HA, and improved the liver pathological variations of fibrotic rats. The experiment proved that the extract of O. caudatum could treat the liver fibrogenesis induced by CCl4 in rats. The positive medicine may inhibit accumulation of extracellular and activate hepatic stellate cell and epithelial-mesenchymal transition.
ABSTRACT
<p><b>OBJECTIVE</b>To testify the TCM theory on "qi-stagnation causes blood stasis" and explore the method of using adrenaline to establish an acute and chronic blood stasis animal model.</p><p><b>METHODS</b>Wistar rats were divided into 5 groups, the blank group (un-modeled), the model group A (modeled with once subcutaneous injection of 0.1% adrenaline), the model group B (modeled with once a day of subcutaneous injection of 0.01% adrenalinei for consecutive seven days), the Salvia group (treated with salvia after modeling) and the Chuanxiong group (treated with Chuanxiong after modeling). The model was established by simulating the anger and anxious status to form acute and chronic blood stasis model respectively and the treatments of Salvia and Chuanxiong was used to testify the syndrome type. Hemorrheologic parameters of the rats were measured.</p><p><b>RESULTS</b>Compared with that in the blank group, the low sheared rate of whole blood viscosity increased, and the RBC deformability and aggregation index raised in both model group A and B (P < 0.01 or P < 0.05). Compared with that in Group A, in the Salvia group, the in vitro thrombus lengthened (P < 0.01), with its dry weight, wet weight increased (P < 0.01) after treatment, but the above-mentioned parameters in the Chuanxiong group were not changed at all.</p><p><b>CONCLUSION</b>One time large dosage and multiple times of small dosage adrenaline subcutaneous injection to simulate angry and anxious status could produce blood stasis animal model. This fact provides an experimental basis for "Qi stagnation causes blood stasis" theory in TCM.</p>