ABSTRACT
Objective To explore the protective effect and mechanism of vitamin D combined with puerarin on liver fibrosis.Methods The rats were divided into normal control group (C),tetrachloromethone group (CCl4),vitamin D group (V),puerarin group(P) and vitamin D combined with puerarin group(V+P).After 8 weeks,the rats were sacrificed and blood and liver samples were collected.The level of blood hyaluronic acid(HA) was tested.The hydroxyproline(Hyp) level in the liver was measured.The liver paraffin sections were made and examined by the sirius red staining.The mRNA levels of collagen Ⅰ and collagen Ⅲ in the liver tissue were detected by RT-PCR,and the levels of NF-κB and TNF-α in the liver were detected by Western blot.Results The CCl4 group appeared obvious liver fibrosis.The liver fibrosis degree was significantly improved in the group V,P and V+P,the blood HA level and liver Hyp level were reduced.The mRNA levels of collagen Ⅰ and collagen Ⅲ as well as the protein levels of NF-κB and TNF-α in the liver were significantly decreased.Among them.The liver fibrosis improvement degree in the V+P group was most significant.Conclusion Vitamin D combined with puerarin can protect rat liver fibrosis induced by CCl4 and its mechanism may be related with reducing the activation of hepatic stellate cells(HSC) and decreasing the collagenous fibers secretion.
ABSTRACT
OBJECTIVE:To study inhibitory effect of Lycianthes biflora polysaccharide on the proliferation of hepatic stellate HSCs-T6 cells in rats and its mechanism. METHODS:After treated with 0(blank control),50,100,200μg/ml L. biflora polysac-charide for 24 and 48 h,the activity of hepatic stellate HSCs-T6 cells in rats was determined by MTT assay and inhibitory rate of cell proliferation was calculated;the content of Hyp in supernatant was detected by immunohistochemical assay. After 48 h,the ex-pression of cellular α-smooth muscle actin(α-SMA)was measured by immunohistochemical assay;both transforming growth fac-tor β1(TGF-β1)and Smad3 were measured by Western blot assay. RESULTS:Compared to blank control,50,100 and 200 μg/ml L. biflora polysaccharide could inhibit the proliferation of HSCs-T6 cells;cell inhibitory rates were 39.84%-69.31% and 45.16%-82.93% respectively after treated for 24 and 48 h,which were positively associated with time and concentration. The con-tents of Hyp in supernatant were 178.36-93.25 μg/ml and 131.94-68.74 μg/ml respectively after treated with different concentrations of PRP for 24 and 48 h,which were negatively associated with time and concentration. The protein level of TGF-β1 and Smad3 de-creased after treated with L. biflora polysaccharide for 48 h(P<0.01). CONCLUSIONS:L. biflora polysaccharide can inhibit the proliferation of hepatic stellate HSCs-T6 cells in rats,and its mechanism is associated with the inactivation of endogenous TGF-βpathway for reducing collagen production.
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Objective To study the effect and mechanism of Hedyotic diffusa injection in reversing multi-drug resistance of BEL-7402/5-FU cells.Methods The IC50 of BEL-7402/5-FU cells was examined by MTT,apoptosis was detected by flow cytometry,and the content of tyrosine kinase in total protein of tumor cells was detected by enzyme-linked immunosorbent assay (ELISA).Results The IC50 of Hedyotic diffusa injection (6 μL/mL ) combined with 5-FU (636.5 μmol/L)decreased significantly compared with that of 5-FU applied alone (1 071.6 6μmol/L). The reversal on BEL-7402/5-FU was 1.68-fold.After injection of 5-FU plus Hedyotic diffusa ,the apoptosis rate of BEL-7402/5-FU cells was increased significantly and the tyrosine kinases of BEL-7402/5-FU cells decreased significantly.Conclusion Hedyotic diffusa injection plays a certain role in reversing multi-drug resistance of BEL-7402/5-FU cells,which may be related to decreasing their tyrosine kinases.