In Vitro Experiment Study Of The Effect Of Valerian Terpene Lactones On Hepatic Stellate Cells Proliferation Induced By Ferric Nitrilotriacetate

Objective:Observed the effect of Valerian terpene lactones on hepatic stellate cells proliferation,and then investigate whether it has an effect on anti-hepatic fibrosis actions.Soprovide adequate theoretical basis for clinical treatment of this disease.Materials and Methods:In vito experiments. Set up four groups namely positive control group, blank controlgroup, model group, drug group. Hepatic Stellate Cells were introduced into ourlaboratory and then subcultured.Select in cell growth logarithmic phase, trypdindigestion, added the culture medium containing10%fetal bovin serum then wind andpercussion into a single cell suspension，and inoculated into96-well plates at a densityof7000-8000cells each well. When nearly adherent monolayer use the mediun (180μL/each well) containing the FeNTA induce cell proliferation except the blankcontrol group. Drug groups also joined of different concentrations(200μg/mL、100μg/mL、50μg/mL、25μg/mL、5μg/mL) ofA、B、C three drugs20μL/each well.Positive control group by adding colchicine20μL (0.5μg/mL). Add the same amountof saline to blank control group and model group. Observe the number andmorphology of the cells under a microscope. Use the MTT colorimetric methord toobserve cell proliferation, and calculate inhibitory rate.Results:The results show that inhibitory rate of Valerian terpene lactones A、B、C (200μg/mL)are40%、36.4%、36.6%, and there were statistically significants compared with model group (P<0.05), OD value (570nm) of Valerian terpene lactones A、B、C (200μg/mL)are0.231±0.014、0.245±0.025、0.244±0.018，and there are significant differencescompared with model group(P<0.01); inhibitory rate of Valerian terpene lactones A、B、C (100μg/mL) are27.5%、21.3%、25.5%, and there are differences compared withmodel group (P<0.05), OD value (570nm) of Valerian terpene lactones A、B、C（100μg/mL）are0.279±0.021、0.303±0.015、0.287±0.015，and there aresignificant differences compared with model group(P<0.01); There is no statisticalsignificance (P>0.05) when compare the OD value of Valerian terpene lactonesA、B、C200μg/mL and100μg/mL dose group with the positive control group; There arestatistically significants when compare the OD value of the model group with thepositive control group (P <0.05). Observe under the microscope: there are more cellsin the model group than in the other groups, and the cells is spindle-shaped orpolygonal or long strip. The cells have good refractive index and there is little cellulardebris and floating cells. But the cell synapses of HSC has disappeared in A、B、C200μg/mL and100μg/mL dose group as well as in the positive group, and the cellsbecome circular while its refraction is poor. Cell debis and floating cells can be seenin the medium. The difference between200μg/mL and100μg/mL dose group is notobvious under microscope.Conclusions:1、Valerian terpene lactones A、B、C in high dose group(200μg/mL) significantlyinhibited the proliferation of hepatic stellate cells; Medium dose group (100μg/mL)also inhibit hepatic stellate cell proliferation.2、With the decline of drug concentration, the inhibition rate decreased,showing acertain degree of dose dependency3、Effect of valerian lactoneA、B、C three drugs in the same concentration is similar...