Study On The Molecular Mechanism Of Patrinia Herba Aqueous Extract And Its Components Induced K562 Cell Apoptosis And Efficacy

Objective: The present study was to explore the potential molecular mechanism of patrinia herba aqueous extract as well as its main component(hederagenin)induced cell proliferation and apoptosis in chronic myeloid leukemia cells(K562 cell),and to search for the potential target of drug induced cell apoptosis.Comparing the effect of them on K562 cells,to provide experimental basis for guiding clinical research and development.Materials and methods: K562 cells were incubated with various concentrations of patrinia herba aqueous extract(2.5mg/ml,5mg/ml,10mg/ml,20mg/ml,40mg/ml)or hederagenin(10umol/l,30umol/l,50 umol/l,70 umol/l,90 umol/l)for 24 h,48h,and 72 h,respectively.Then the cell vitality was measured by MTT assay and the inhibitory rate was calculated to obtain IC50 by Graphpad software.According to the IC50 values,the appropriate drug concentration and incubation time was chosen to act on K562 cells and cell apoptosis was detected by flow cytometry.Finally the relative expression of STAT3,c-fos,c-jun gene was detected by q RT-PCR.Results: 1.MTT assay results showed that both patrinia herba aqueous extract could significantly inhibit the proliferation of K562 cells,and the similar inhibition effect was also observed when K562 cells were incubated with hederagenin.The inhibitory effect was both time and concentration dependent.With the increase of drug concentration and the extension of time,the inhibition of proliferation was enhanced.When incubated with the above concentrations of hederagenin,the IC50 values of 24 h,48h and 72 h in K562 cells were84.74umol/l,71.75 umol/l and 48.43 umol/l,respectively.When incubated with the above concentrations of patrinia herba aqueous extract,the IC50 values of 24 h,48h and 72 h in K562 cells were 24.77mg/ml,15.88 mg/ml and 10.73 mg/ml,respectively.2.The results of flow cytometry showed that hederagenin not only promotes cell apoptosis,which the proapoptotic effect increases with the increase of drug concentration.but also makes cell death(P<0.001,compared with control).When K562 cells were incubated with patrinia herba aqueous extract,we can observed that the low concentrations(2.5mg/ml,5mg/ml)of patrinia herba aqueous extract had little effect on the apoptosis of K562 cells(P>0.05,compared with control);However,high concentrations(10 mg/ml,20 mg/ml,40 mg/ml)of patrinia herba aqueous extract significantly promoted the apoptosis of K562 cells(P < 0.001,compared with the control group),and the proapoptotic effect increases with the increase of drug concentration.3.The relative expression of STAT3,c-fos,and c-jun gene in K562 cells after treatment was detected by q RT-PCR.The results showed that the expression of STAT3 gene was significantly decreased(P < 0.001)after 10mg/ml patrinia herba aqueous extract or 70umol/l hederagenin,respectively,and the expression of c-jun and c-fos was significantly increased in K562 cells(P<0.05).4.Patrinia herba aqueous extract and its main components of hederagenin could significantly inhibit the proliferation of K562 cells and induced cell apoptosis,the expression of STAT3 gene was significantly decreased,while the expression of c-jun and c-fos was increased in K562 cells.what is more,When the same effect was achieved on inhibiting the proliferation of K562 cells and induceding cell apoptosis,hederagenin with less,the effect was good.Conclusions: 1.Patrinia herba aqueous extract and its main components of hederagenin could significantly inhibit the proliferation of K562 cells and induced cell apoptosis,which the inhibitory effect was both time and concentration dependent.With the increase of drug concentration and the extension of time,the inhibition of proliferation was enhanced.2.Patrinia herba aqueous extract and hederagenin might inhibit K562 cell proliferation and promote cell apoptosis by decreaseding the expression of STAT3 gene and increaseding the expression of c-jun and c-fos.3.Patrinia herba aqueous extract play anti-tumor effect by its main component hederagenin.4.When the same effect was achieved on inhibiting the proliferation of K562 cells and induceding cell apoptosis with Patrinia herba aqueous extract and its main components of hederagenin,the latter with less,the effect was good.Which suggests that the latter is more effective on playing anti-tumor role.