ABSTRACT
Objective To observe the effect of TGF-β1 on activation and epithelial mesenchymal transition(EMT) in rat hepatic stellate cell-T6.Methods Adopt the MTT method to screen the optimum concentration of TGF-β1 in vitro HSC-T6 cultured.After the HSC-T6 stimulation by TGF-β1 of 10 μg/L for 24 hours, the morphology of the cells was observed under inverted phase contrast microscope, the expression of F-actin which on behalf of cotoskeletal structure was detected by immunofluorescence staining;the expression of α-SMA and N-cadherin,vimentin,E-cadherin was measured by RT-qPCR;The changes of α-SMA,N-cadherin,vimentin and E-cadherin were assessed by Western blot after different concentrations (0,5 and 10 μg/L) of TGF-β1 interventing HSC-T6 for 24 h.Results The optimal cell survival rate was recorded when 10 μg/L TGF-β1 dealt withcells for 24 h.After HSC-T6 were treated with TGF-β1,cells stretched, pseudopodia increased and turn into stellate, cells connections were looser, so that represented a significantly activated state.F-actin filaments gathered to form stress and distributed along the long axis of the cells;The expression of α-SMA mRNA and vimentin mRNA in experimental group was significantly higher while E-cadherin mRNA was obviously lower than the control group(P<0.05).TGF-β1 made the protein expression of α-SMA and N-cadherin, vimentin in dose-dependent increased while E-cadherin was decreased.Conclusions TGF-β1 may induce activation and epithelial-mesenchymal transition of HSC-T6.
ABSTRACT
AIM: To establish a HPLC method for determining notoginsenoside R_1, ginsenoside Rg_1, Rb_1 and Rd in Suxiong Pill (Radix et Rhizoma Notoginseng, Flos Carthami, Rhizoma Chuanxiong). METHODS: A Kromasil C_ 18 column was used. The mobile phase was acetonitrile-water in linear gradient elution. The detection wavelength was 203 nm. RESULTS: The linear range was at 1.20-6.02 ?g(r= 0.999 6 ) for notoginsenoside R_1, 5.60-27.99 ?g(r= 0.999 9 ) for ginsenoside Rg_1, 2.84-14.20 ?g(r= 0.999 9 )for ginsenoside Rb_1, 0.51-2.55 ?g(r= 0.999 7 ) for ginsenoside Rd. The average recoveries (n=6) were 96.4% (RSD= 1.9% ) , 103.7% (RSD= 0.8% ), 106.6% (RSD= 1.3% ), 98.5% (RSD= 1.1% ), respectively. CONCLUSION: The method is simple, reliable, accurate and can be applied to the quality control of the preparation.