Feng Lili, Cao Wenfu, Ye Feng. Mechanism of Ezhu-containing serum inhibiting the expression of Gli1 and β-catenin in hepatic stellate cells. [J]. Journal of Chongqing Medical University 42(3):308-314(2017)
DOI:
Feng Lili, Cao Wenfu, Ye Feng. Mechanism of Ezhu-containing serum inhibiting the expression of Gli1 and β-catenin in hepatic stellate cells. [J]. Journal of Chongqing Medical University 42(3):308-314(2017) DOI: 10.13406/j.cnki.cyxb.001221.
Mechanism of Ezhu-containing serum inhibiting the expression of Gli1 and β-catenin in hepatic stellate cells
Objective:To explore the mechanism of Ezhu-containing serum inhibiting the expression of glioma-associated oncogene homolog-1(Gli1
the nuclear transcription factor of hedgehog signaling pathway)and β-catenin(the core factor of wnt signaling pathway)in hepatic stellate cells(HSC)stimulated by leptin. Methods:Thirty sprague-dawley(SD) rats were random Ly divided into 3groups(n=10)
which were treated with Ezhu-decoction
colchicine and physiological saline by gavage for 10 d in order to prepare drug-containing serums. The method of high performance liquid chromatography(HPLC)was used to detect the composition of Ezhu in serum. The HSC during the exponential growth phase were divided into 8 groups:blank control group
model group
hedgehog pathway inhibitors(cyclopamine) group
ezhu group
Ezhu and cyclopamine group
hedgehog pathway agonost(pumorphamine)group
Ezhu and purmorphamine group
colchicine group. HSC were cultured in vitro and stimulated with 100 ng/m L leptin(except for the blank control group)
then were treated separately with the corresponding drug for 24 h. After cells being collected
HSC proliferation was detected using MTT colorimetric assay;the expressions of Gli1 and β-catenin were determined by PT-PCR
Western blot and immunofluorescence
respectively. Results:Compared with those of blank control group
the expressions of Gli1 m RNA and Gli1 proteins and β-catenin m RNA and β-catenin proteins were obviously increased after stimulating the HSC of rats by leptin(All P=0.000). Compared with those of model group
the expressions of Gli1 m RNA and Gli1 proteins and β-catenin m RNA and β-catenin proteins were decreased obviously(All P=0.000)after being interfered by Hh pathway(cyclopamine)and ezhu-containing serum. Compared with the model group
using ezhu-containing serum to interfere the Hh pathway inhibitors group can make the expression of Gli1 m RNA and Gli1 proteins and β-catenin m RNA and β-catenin proteins decreased obviously(All P=0.000). Compared with those of the model group
after being interfered by colchicines
the expressions of Gli1 m RNA and Gli1 proteins were decreased obviously(P=0.000
P=0.007)
and β-catenin m RNA andβ-catenin proteins were decreased obviously(All P=0.000). Compared with those of model group
the expressions of Gli1 m RNA and Gli1 proteins and β-catenin m RNA and β-catenin proteins were increased obviously(All P=0.000)after adding Hh pathway agonist(purmorphamine). Compared with those of purmorphamine group
the expressions of Gli1 m RNA and Gli1 proteins and β-catenin m RNA and β-catenin proteins decreased obviously(All P=0.000)after using ezhu-containing serum to interfere the purmorphamine group. Conclusion:Ezhu-containing serum plays an important part in inhibition of HSC activation and increment by taking part in hedgehog signaling pathway to regulate the expression of Gli1. Ezhu-containing serum could inhibit the expression of β-catenin(the key factor of Wnt signaling pathway)by regulating the expression of Gli1 in leptin-induced HSC