ABSTRACT
OBJECTIVE To investigate the toxicological effect of patulin(PAT)on the growth of human normal liver cells L-02 and its possible mechanisms. METHODS After cells were treated with PAT 1.25, 2.5,5,10 and 20μmol·L-1 for 24 or 48 h,cell viability was examined using MTT assay. L-02 cells were treated with PAT 5 and 10 μmol · L- 1 for 24 h ,respectively. Cytomorphology and mitochondrial membrane potential (MMP) were observed under a fluorescence microscope. Apoptosis,MMP and reactive oxygen species (ROS)were analyzed by flow cytometry. Mitochondria apoptosis pathways were detected by Western blotting. RESULTS PAT exhibited a strong inhibitory effect on L-02 in a concentration-dependent and time-dependent manner. IC50 of PAT treatment for 24 or 48 h was 6.61 and 2.78 μmol · L-1,respectively. MMP was decreased,while the percentage of low MMP cells increased from(9.2±2.3)%in controls to(23.4±4.5)%( PAT 5μmol·L-1)and(47.1±5.5)%(PAT 10μmol·L-1), respectively. Compared to untreated cells,the early apoptosis population increased from(3.8±1.1)%to(29.8±4.5)%( PAT 5μmol·L-1)and (24.1±6.2)%(PAT 10μmol·L-1)(P<0.01),respectively. Further?more,the accumulation of ROS was also observed. The effect of PAT on ROS and cell viabilities could be attenuated by glutathione. CONCLUSION PAT can significantly inhibit the growth of L-02 and induce apoptosis via ROS-dependent mitochondria pathways.
ABSTRACT
Objective To investite the effect of endotoxin pretreatment on lung injury induced by hepatic ischemia reperfusion in rabbits and its mechanism. Method Forty-eight New Zealand white rabbits were randomly divided into4 groups with 12 rabbits each group:routine control group,pretreatment control group,ischemia reperfusion group (IR group), and preperfusion group( LPS + IR group). Rabbits of routine control group received operative dissector only, and those of pretreatment control group received pretratment of daily intraabdominal injection of lipopo|ysaccharide(O.5,0.5,and 1.0 mg/kg,respectively)in the 3 days before operative dissector.Livers of IR group were rendered and ischeraic for 30 minutes, and repeffused for up to 4 hours. Rabbits of LPS +IR group received the preueaunent before heretic ischemia repeffusion. Four hours after reperfusion, serum endotoxin,tumor necrosis factor-α(TNF-α), wet/dry ratio and broncho-alveolar lavage fluid protein content of lung,malondialdehyde(MDA) and mpenrxide dismutase(SOD) in lung homogenate, lung injury ratio, and activity of Nuclear factor-kB(NF-kB) in alveolar macrophage wene examined. Differences within the groups were analyzed using One way ANOVA. Results Between the two control groups,there were no significant differences in all indexes(P>0.05). The TNF-α[ (48.31±5.31)pg/ml vs.(56.47±5.09)pg/ml, P<0.01],wet/dry ratio [(4.98±0.33)vs. (5.22±0.31), P = 0.03],broncho-alveolar hvage fluid protein content[(0.68±0.11)g/L vs. (0.76±0.10)g/L, P =0.04],MDA[(0.86±0.06)nmol/mg vs. (0.93±0.07)nmol/mg, P =0.02],lung injury ra-tio[(13.4±4.3)% vs. (17.4±4.1)%, P = 0.03],and the activity of NF-gB[(5.82±1.12)OD/mm2 vs.(7.40±1.26)OD/mm2, P<0.01] in alveolar macrophage of the LPS+ IB group were all significantly lower than those of IB group, while the SOD[ (90.30±7.38 )U/rag vs. (84.44±7.90 )U/rag, P = 0.04]of LPS + IR group was significantly higher than that of IR group. Conclusions Endotoxin pretrealment may ameliorate the lung injury induced by hepatic isehernia reperfusion. The mechanism may be that endotoxin pretreatment deoreases production of serum TNF-α and the activity of NF-kB in alveolar maerophage.