Effects Of Danshen-containing Serum On Expression Of Correlation Factor Of Hedgehog Signaling Pathway In HSCs

ObjectiveTo discuss the effect of Danshen-containing serum on expression of correlation factor of Hedgehog signaling pathway of Rat HSCs stimulated by leptin.Methods60 SD rats were randomly divided into 3 groups(n=20),which were treated with Danshen Decoction,Physiological saline and Colchicine by using the way of intragastric administration for 10 days in order to prepare medicated serum. Determination of main components of tanshinol and tanshinone IIA in the Danshen-containing serum by HPLC. To divide the HSCs in logarithmic growth phase into seven groups: Blank control gruop, Model group, Cyclopamine group, Danshen group, Purmorphamine group, Purmorphamine+Danshen group, Colchicine group. Except for Blank control group, other groups were stimulated for 24 hours by providing 100ng/m L leptin. After intervened by the drug serum, each group was cultured at 37℃in 5% CO2. Cells were collected after 24 hours, then the MTT Colorimetry was used to test Danshen-containing serum’s effect on the proliferation of HSCs; Real-time PCR used to test the m RNA expression of Smo, Su Fu, DYRK2, EVC2, Gli2 and α-SMA; The Western-Blot method used to test the protein expression in Smo, Su Fu, DYRK2, EVC2, Gli2, α-SMA and the Immunofluorescence method used to test the expression of EVC2 in HSCs of each group.ResultsAfter the stimulation by Leptin to rats, the expression of Smo, DYRK2, EVC2, Gli2, α-SMA in HSCs were increased more obviously than that in Blank control group, while the expression of Su Fu is much lower compared with Blank control group(P < 0. 05 or P < 0. 01). After the intervention of Cyclopamine, Danshen-containing serum and Colchicine, the expression of Smo, DYRK2, EVC2, Gli2, α-SMA decreased much more than that in Model group, while the expression of Su Fu obviously increased much more in comparison with that in Model group(P < 0. 05 or P < 0. 01). Based on the Model group, the Purmorphamine was added and fully activated, the expression of Smo, DYRK2, EVC2, Gli2, α-SMA were significantly higher than those in Model group, while the expression of Su Fu is much lower than that in Model group(P < 0. 01). After Danshen-containing serum intervent the Purmorphamine group, the expression of Smo, DYRK2, EVC2, Gli2, α-SMA were significantly lower than those in Model group, while the expression of Su Fu increased much more obviously than that in Model group(P < 0. 01).ConclusionDanshen-containing serum can affect the expression of correlation factor of Hedgehog signaling pathway in HSCs by leptin intervention, and then it impacts on the process of the activation and proliferation of HSCs.