ABSTRACT
OBJECTIVE To establish the fingerprint of Sophora flavescens， and to screen differential components and determine their contents. METHODS HPLC fingerprints of 12 batches of S. flavescens were established by using Similarity Evaluation System of Chromatographic Fingerprints of TCM （2012 edition）； common peaks were identified and their similarities were evaluated. Chemical pattern recognition analysis [cluster analysis （CA），principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）] were performed with SIMCA 14.1 and SPSS 23.0 software， and differential components which influenced the quality of S. flavescens were screen with variable importance in the projection（VIP）＞1 as standard. Meanwhile， the contents of 4 kinds of differential components were determined by the same HPLC method. RESULTS There were 17 common peaks in the fingerprints of 12 batches of S. flavescens，and their similarities were all higher than 0.96. A total of 6 common peaks were identified， i.e. oxymatrine （peak 1）， oxysophocarpine （peak 2）， matrine （peak 10）， trifolirhizin （peak 14）， kurarinone （peak 16） and norkurarinone （peak 17）. Results of CA， PCA and OPLS-DA showed that 12 batches of S. flavescens were divided into 3 categories according to producing area， i.e. S1-S7 （Shangzhou District of Shaanxi Province） were grouped into one category， S8-S10 （Yichuan County of Henan Province） into one category and S11-S12 （Chifeng City of Inner Mongolia） into one category. VIPs of matrine， norkurarinone， kurarinone and oxysophocarpine and the chemical components represented by peak 11 and 9 were all greater than 1. The contents of matrine， norkurarinone， kurarinone and oxysophocarpine in 12 batches of S. flavescens were 2.65-4.93， 1.54-3.44， 9.63-12.94 and 5.08-6.10 mg/g， respectively. CONCLUSIONS HPLC fingerprint of S. flavescens is established successfully in the study， and can be used to screen 6 differential components by combining with chemical pattern recognition analysis， which can provide reference for quality control of S. flavescens.
ABSTRACT
Objective To evaluate the effect of endoplasmic reticulum stress in trophocytes,in patients with intrahepatic cholestasis of pregnancy (ICP).Methods Sixty-one pregnant women who were hospitalized in Women's Hospital,School of Medicine,Zhejiang University from January to December 2015 were recruited.Thirty-one women who were diagnosed as ICP were defined as the ICP group and 30 healthy pregnant women were defined as the control group.The localization and expression intensity of glucose regulated protein 78 (GRP-78) in placental tissues were detected by immunohistochemistry technique.Electronic microscope was used to observe ultra-microstructure change of the endoplasmic reticulum in trophocytes and cell line Swan71.Reverse transcription (RT)-PCR and western blot were used to investigate the expression of GRP-78 mRNA and protein in Swan 71 cell.Results (1) GRP-78 protein was mainly expressed in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts.The protein expression of GRP-78 in placentas of the ICP group (13.2±2.4) was significantly higher than that in the control group (7.8±1.3,P＜0.01).(2) The volume of endoplasmie reticulum did not increase and the microvilli developed well,with no swelling and no expansion of endoplasmic reticulum in the control group.In the ICP group,microvilli injury,endoplasmic reticulum edema were found;the volume of endoplasmic reticulum increased,with dilation,vacuolation and significant degranulation.After treated with 100 μmol/L cholyglycine for 24 hours,universal dilatation of the endoplasmic reticulum were seen in the Swan71 cells.(3) In Swan71 cells,cholylglycine displayed a concentration-dependent up-regulation on the expression of GRP-78.The expressions of GRP-78 mRNA in 0,25,50,100 μmol/L cholylglycine experimental group were 1.01±0.17,2.17±0.16,5.47±0.36,5.65 ± 0.82,respectively.The expression of GRP-78 protein in 0,25,50,100 μmol/L cholylglycine experimental group were 1.01±0.04,1.17±0.15,1.33±0.13,1.73±0.13,respectively.The expression of GRP-78 mRNA and protein in 100 and 50 μ mol/L cholylglycine experimental group were significantly higher than 0 μmol/L (all P＜0.01).Conclusion The obvious expansion of endoplasmic reticulum and the increased expression of GRP-78 in trophocytes indicated that endoplasmic reticulum stress of trophocytes may be involved in the pathogenesis of ICP.