ABSTRACT
OBJECTIVE： To investigate the effects of chelidonine on proliferation， collagen synthesis and TGF-β1 receptor of activated hepatic stellate cells CFSC-8B. METHODS： CFSC-8B cells in logarithmic phase were collected and then divided into normal control group， model group， solvent group （ethanol）， positive control group （1 μg/mL colchicine ethanol solution）， chelidonine low， medium and high concentration groups （2.1， 4.2， 8.4 μg/mL chelidonine ethanol solution）. Except for normal control group， other groups were activated with 20 μg/L TGF-β1 for 24 h； the latter 5 groups were intervened with relevant medicine for 24 h. Cell proliferation of activated cells was assayed by CCK-8 assay. Hydroxyprolin （Hyp） content was assayed by enzyme digestion； the levels of typeⅠ collagen （Col-Ⅰ） and type Ⅲ collagen （Col-Ⅲ） were assayed by ELISA； the expressions of TβR-Ⅰ and TβR-Ⅱ protein were assessed by Western blot； mRNA expressions of α-SMA， TβR-Ⅰ and TβR-Ⅱ in hepatic stellate cells were assessed by RT-PCR. RESULTS： Compared with normal control group， cell proliferation rate， Hyp content， the levels of Col-Ⅰ and Col-Ⅲ， the protein expressions of TβR-Ⅰ and TβR-Ⅱ as well as mRNA expressions of α-SMA， TβR-Ⅰ and TβR-Ⅱ were increased significantly （P＜0.05）. Compared with model group， there were no significant difference in above indexes of hepatic stellate cells in solvent group （P＞0.05）； there were no significant difference in the proliferation rate of hepatic stellate cells in chelidonine low concentration group （P＞0.05）， above indexes of hepatic stellate cells were decreased significantly in positive control group and chelidonine high concentration group （P＜0.05）. The decrease of Hyp and Col-Ⅲ levels were not significant in chelidonine medium concentration， but other above indexes were decreased significantly （P＜0.05）. Compared with chelidonine medium concentration group， the rate of cell proliferation， Col-Ⅰ level， protein and mRNA expressions of TβR-Ⅰ and TβR-Ⅱ were decreased significantly in chelidonine high concentration group （P＜0.05）. CONCLUSIONS： Chelido- nine can inhibit the proliferation， collagen synthesis as well as the protein and mRNA expressions of TβR-Ⅰand TβR-Ⅱ in activated CFSC-8B cells.
ABSTRACT
Curdione is a main active component of curcuma rhizomes [Ezhu], which shows an excellent antithrombotic activity. In this study, the concentration of Curdione in pregnant rats and their offspring brain was determined using ultra-performance liquid chromatography-quadrupole time-of-flight/mass spectrometry [UPLC-Q-TOF-MS] method. The water extraction then alcohol precipitation extract from Ezhu was administered through tail intravenous injection. The pharmacokinetic parameters were analyzed to compare the differences between the blood stasis group rats and normal group rats. Using Schisandrol A as an internal standard, samples were extracted using dichloromethane and isopropanol [90:10, v/v]. Calibration plot was linear over the range of 0.5-200micro g·mL[-1] for Curdione in brain with the lower quantification limit being 0.5micro g·mL[-1]. The recoveries of Curdione and IS from brain were more than 93.31% and 90.90% separately. The RSD for both intra- and inter-day precision were <6.49%, RE were -14.84%[tilde]-2.8%. The pharmacokinetic parameters C[max] and AUC among the four kinds of rats had significant difference. The Curdione distributed in rat brain in model group is less than normal group. Ezhu medicine may show the therapeutic effect but not the reproductive toxicity on mother or unborn baby to cure the pregnant women under the adaptive symptoms
ABSTRACT
This paper was aimed to study the protective effects and related mechanisms of Zhen-Gan Xi-Feng (ZGXF) decoction containing serum on 6-OHDA-induced oxidative stress in PC12 cells.The ZGXF decoction containing rat serum with low-,medium-,and high-dose (8,16,or 32 g.kg-1) or blank serum was used to preprocess PC12 cells for 1 h,and cultured together with 100 μM 6-OHDA for 24 h.And then,cells were collected.The fluorescent probe 2',7'-dichlorofluorescin diacetate (DCFH-DA) and fluorescence microplate reader were used to detect the level of reactive oxygen species (ROS).Real-time quantitative PCR was used to analyze the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2),Nfe2l2,heme oxygenase-1 (HO-1),glutamate-cysteine ligase catalytic subunit (GCLc),and GCL modulatory subunit (GCLm).The luciferase report gene system was used to detect the antioxidant response element (ARE) activation.The results showed that ZGXF decoction-containing serum inhibited the 6-OHDA-induced oxidative stress,upregulated the Nfe2l2,HO-1 and GCLc mRNA expressions in cells processed with 6-OHDA.However,it has no significant effect on GCLm mRNA expression.It was concluded that ZGXF decoction-containing serum had protective effects on 6-OHDA-induced oxidative stress in PC 12 cells.Its mechanism may be correlated with the upregulation on Nfe2l2 mRNA expression,which activated ARE and further induced its downstream gene of phase Ⅱ detoxifying enzyme,as well as the HO-1 and GCLc mRNA expressions of antioxidant enzyme gene.
ABSTRACT
The aim of this study was to explore the effects of ZGXF decoction on amphetamine-induced rotation in PD rats with syndrome of liver-yang hyperactivity,and its mechanism involved.Rats received 6-OHDA administration via intra-substantia nigra injection and were intragastrically treated by Fu Zi decoction to establish the PD model with the syndrome of liver-yang hyperactivity.Three doses of ZGXF decoction or selegiline were given by a 28-day intragastric administration.The rats were tested for amphetamine-induced rotation asymmetry.In addition,real-time PCR were adopted to analyze the expressions of Nfe212 and Hmox1 mRNAs,while western blot to analyze the expression of Keap1 protein.As a result,it was found that ZGXF decoction dose-dependently attenuated amphetamine-induced rotation,up-regulated the expressions of Nfe212 and Hmox-1 mRNAs,and down-regulated the expression of Keap1 protein in the substantia nigra in PD rats with syndrome of liver-yang hyperactivity.It was suggested that anti-PD effects of ZGXF decoction be attributed to the up-regulation of Nfe212 and Hmox-1 mRNAs and the down-regulation of Keap1 protein,being associated with oxidative stress,in the substantia nigra of PD rats with syndrome of liver-yang hyperactivity.