The Effect Of Valerian Terpene Lactones On Hepatic Stellate Cells Proliferation And Expression Of Type ⅠCollagen And TGFβ1

Objective:To investigate the effect of Valerian terpene lactones on hepatic stellate cells proliferation and expression of type I collagen and TGFβ1induced by Ferric nitrilotriacetate in vitro.Materials and Methods:Select cell in growth logarithmic phase, then digest and wind and percussion the cell into suspension, and inoculated into96-well plates at a quantity of1×104cells/well. When cultured to monolayer adherent, decant the old medium and use the medium containing3%fetal bovine serum. Set up four groups namely control group, model group, positive control group and drug group (valerian terpene lactone A, B, C group,each include five concentration of50ug/ml,100ug/ml,200ug/ml,300ug/ml,400ug/ml). Add the0.1mmol/L FeNTA (10ul/well) to each well except the blank group, and the drug group were added by Valerian terpene lactones A, B, C (10ul/well)of different concentrations. Positive control group were joined of colchicine (0.5ug/ml, lOul/well). The same amount of saline was added to blank control group and model group. The total amount of each hole is100ul. After interaction with drug for36h, observe the number and morphology of the cells under a microscope, and check the proliferation of the HSC by MTT colorimetric assay. Divided the cells into blank control group, model group, positive control group and valerian terpene lactones C (50ug/ml,100ug/ml,200ug/ml,300ug/ml,400ug/ml) group with the same method as above. Taken culture supernatant after36h, and test the content of collagen I and TGFβ1by using the ELISA. Results:1. The result show that the growth inhibition rate of Valerian terpene lactones A400ug/ml,300ug/ml and200ug/ml groups are34.6%,29.3%,19.4%, they can significantly inhibit the proliferation of HSC compared with model group in vitro culture (P<0.01);The inhibition rate of Valerian terpene lactones B400ug/ml,300ug/ml,200ug/ml and100ug/ml groups are44.6%,35.2%,23.4%,13.7%,there are significant difference compared with the model group. Inhibitory rate of Valerian terpene lactones C400ug/ml,300ug/ml,200ug/ml,100ug/ml and50ug/ml groups are51.9%,43.2%,30.1%,20.2%,10%,when compared with the model group, it is significant difference (P<0.01). There are not statistically significant of other concentration groups (P>0.05). When compared with the positive control group, the difference of A (400ug/ml) group, B (400ug/ml the B300ug/ml) groups andC (400ug/ml, C300ug/ml C200ug/ml) groups are significant (P<0.01); Compare A (300ug/ml) and B (200ug/ml) group with the positive control group, the difference are statistically significant (P<0.05), and it show that the inhibition rate of above-mentioned groups are greater than colchicine,while the other concentration groups are lower (P<0.01). There is significant difference when compare the OD value of the model group with the positive control group (p<0.01).2. After interact with HSC, Valerian terpene lactones C400ug/ml and300ug/ml groups (the content of type I collagen are90.86±7.62ng/L and119.49±3.32ng/L) can significantly inhibit the expression of type I collagen contrast with model group (169.64±12.01ng/L)(p<0.01), and there is statistically significant of200ug/ml group (138.32±8.05ng/L)(p<0.05). It’s no statistically significant of100ug/ml and50ug/ml groups compared with model group (p>0.05).Compared with the positive control group(106.95±4.31ng/L), the secretion amount of type I collagen of400ug/ml group was significantly higher than colchicine (p<0.01), while300ug/ml group have meaningless difference (p>0.05).The secretion of type I collagen of 200ug/ml,100ug/ml and50ug/ml groups are significantly less than colchicine group (p<0.01).3. After interact with HSC, Valerian terpene lactones C400ug/ml and300ug/ml groups (the content of TGFβ1are119.62±7.27pg/ml and137.78±4.03pg/ml) can significantly inhibit the expression of TGFβ1contrast with model group (155.51±5.56pg/ml)(p<0.01), and there is statistically significant of200ug/ml group (147.04±3.67pg/ml)(p<0.05). It’s no statistically significant of100ug/ml and50ug/ml groups compared with model group (p>0.05). Compared with the positive control group (143.88±2.21pg/ml), the secretion amount of TGFβ1of400ug/ml group was significantly higher than colchicine (p<0.01), while300ug/ml group have meaningless difference (p>0.05).The secretion of TGFβ1of200ug/ml,100ug/ml group (p<0.05) and50ug/ml group (p<0.01) are significantly less than the colchicine group.Conclusions:1. The inhibitory effect of the valerian terpene lactones A, B, C in different concentrations on proliferation of hepatic stellate cell is not alike. With the drug concentration increasing, the inhibition rate increased, showing a certain dose dependency. It’s C>B>A of drug efficacy at the same concentration except A and B which have similar efficacy at50ug/ml.2. Valerian terpene lactones C (400ug/ml,300ug/ml and200ug/ml) can decrease the production of type I collagen.3. Valerian terpene lactones C (400ug/ml,300ug/ml and200ug/ml) can decrease the production of TGFβ1, thereby weaken the secretion effect of HSC.