Effects And Mechanism Of Danshen Drug-Containing Serum On Proliferation In Hepatic Stellate Cells

ObjectiveTo investigate the effects of Danshen drug-containing serum on proliferation and expression of retinoic acid receptor in hepatic stellate cells,and the mechanism on TGF-β1 and Cyclin D1 in hepatic stellate cells.MethodsAccording to the random number table,30 Sprague-Dawley(SD) rats were randomly divided into the Danshen(1.04 g /kg, 2.08 g /kg, and 4.17 g /kg dosage) groups, colchicine group and normal control group. Blood was collected after gastric perfusion at 2 hours on the 10 th day. The best concentration of PDGF-BB to promote proliferation of HSCs was Determined by MTT, while the effecting of Danshen drug-containing serum on proliferation of HSCs was measured by CCK-8.The method of high performance liquid chromatography(HPLC) was used to detect Salvianic acid and Tanshinone IIA in the Danshen drug-containing serum. HSCs were cultured in vitro and stimulated with PDGF-BB, then were treated separately with the serum of Danshen(high, middle, and low dosage), normal saline for 24 hours. After cells being collected, the expression of retinoic acid receptor and TGF-β1 m RNA were measured by real-time PCR(q RT-PCR), the content of a-SMA, typeⅠcollagen, RXR-α and Cyclin D1 were detected by Western blot.Results1. HPLC showed that the chromatographic peak of Salvianic acid and Tanshinone IIA appears in the Danshen drug-containing serum group.2.MTT showed that with the concentration of PDGF-BB increasing, cell survival rate increased significantly, especially at the concentration of 10 ng/ml.3.CCK-8 showed that compared with model group,Danshen(1.04 g/kg) drug-containing serum reduced the cell survival rate(P < 0. 05); Danshen(2.08 g/kg,4.17 g/kg) drug-containing serum decreased the cell survival rate significantly(P < 0. 01).4.Compared with the normal control group, Danshen(low, middle, and high dosage) drug-containing serum could increase the expression of retinoic acid receptor m RNA, and decrease the expression of TGF-β1 m RNA, a-SMA, typeⅠcollagen and Cyclin D1(P<0.05),and in the middle,high dose groups, the trend was more obvious(P<0.01). Compared with the colchicine group, Danshen(middle and high dosage) drug-containing serum could reduce the contents of typeⅠcollagen and increase the RXR-α,then a-SMA decreased in the Danshen high dosage drug-containing serum(P < 0. 05).ConclusionPDGF-BB can promote the activation of hepatic stellate cells;Danshen can inhibit the proliferation of HSCs, it may be achieved through increasing the expression of retinoic acid receptor and reducing TGF-β1 and Cyclin D1 to inhibit the proliferation of HSCs, thus play a role in the inhibition of liver fibrosis.