Protective effect of fructus broussonetiae on APAP-induced injury of L02 cells
Objective To investigate the protective effect of fructus broussonetiae on L02 cell damage induced by acetaminophen(APAP).Methods The survival rate of normal L02 cells was determined by CCK-8 method at different concentrations(0.1,0.3,1.0,3.0,10.0 mg/ml).After the L02 cells were infected with APAP,the L02 cells with logarithmic growth stage were selected and divided into normal group,APAP group(40 mmol/L)and fructus broussonetiae group(1 mg/ml).The cell survival rate was determined by CCK-8 method,and the intracellular ROS levels were determined by DCFA-DH method.Apoptosis was observed by Hoechst 33258 staining.The expression of mitochondrial membrane potential in L02 cells was detected by Rhodamine 123 staining.The expression levels of JNK,p-JNK,78GRP78,and CHOP were detected by Western blot.Results Compared with 0.1 mg/ml fructus broussonetia water extract,the cell activity of 0.3,1.0,3.0,and 10.0 mg/ml fructus broussonetia water extract decreased,and the difference was highly statistically significant(P＜0.01).Compared with normal group,the cell survival rate and the average fluorescence intensity of apoptosis of APAP group were decreased(P＜0.05).Compared with APAP group,the survival rate of 1 mg/ml fructus broussonetia water extract increased,and the average fluorescence intensity of apoptosis decreased(P＜0.05).The ROS fluorescence inten-sity of APAP group was higher than that of normal group,and the differences were highly st atistically significant(P＜0.01).The ROS fluorescence intensity of fructus broussonetiae seed group was lower than that of APAP group,and the differences were highly statistically significant(P＜0.01).The average fluorescence intensity of mitochondrial membrane potential in APAP group was lower than that in normal group,and the difference was highly statistically significant(P＜0.01).The average fluorescence intensity of mitochondrial membrane potential in fructus broussonetia group was higher than that in APAP group,and the difference was highly statistically significant(P＜0.01).There was no significant difference in JNK protein expression among all groups(P＞0.05).Compared with normal group,the expression levels of P-JNK,GRP78,and CHOP protein in APAP group were increased(P＜0.01).Compared with APAP group,the protein expression levels of P-JNK,GRP78 and CHOP in fructus broussonetiae group were decreased(P＜0.01).Conclusion Fructus broussonetiae mitigates liver injury by inhibiting JNK,restoring mitochondrial membrane permeability,suppressing endoplasmic reticulum stress,lowering ROS levels,and inhibiting apoptosis.
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