[Relationship between copper injury and apoptosis and the effect of curcumin on copper-injured BRL cells].
OBJECTIVE
To study the relationship between copper-induced apoptosis and reactive oxygen species (ROS) in BRL cells and the effect of curcumin, a plant-derived polyphenol, on copper-injured BRL cells.


METHODS
BRL cells were treated with CuSO4 (100 micromol/L) or curcumin + CuSO4. The BRL cells without any treatment were used as controls. Flow cytometry was applied to detect the production of ROS with fluorescent probe DCFH-DA. MTT colorimetry was used to evaluate cell activity. Apoptosis was measured using Hoechst 33258 staining and Annexin V-FITC and propidiumiodide (PI) staining. JNK/SAPK protein level was detected using Western blot.


RESULTS
ROS levels (711.70 +/- 68.33 vs 87.22 +/- 7.58) and apoptosis rate (45.08 +/- 1.87% vs 8.23 +/- 2.56%) of BRL cells reached to a peak after 6 hrs of CuSO4 treatment, which were significantly higher than controls (P < 0.01). JNK/SAPK levels increased significantly after 6 hrs of CuSO4 treatment and peaked at 24 hrs of CuSO4 treatment compared with controls (P < 0.01). Curcumin pretreatment decreased significantly ROS and JNK/SAPK levels as well as the apoptosis rate when compared with the CuSO4-treated alone group (P < 0.01).


CONCLUSIONS
Copper may induce apoptosis of BRL cells. ROS participated in apoptosis induced by copper. Curcumin produced protections on copper-injured BRL cells possibly by anti-oxidation and inhibition of p-JNK expression.
Wan
Xiao-Hua
XH
Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Luo
Xiao-Ping
XP
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002460:Cell Line; D003300:Copper / Q000633:toxicity*; D003474:Curcumin / Q000494:pharmacology*; D005434:Flow Cytometry; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D008099:Liver / Q000187:drug effects* / Q000473:pathology; D010766:Phosphorylation; D051381:Rats; D017382:Reactive Oxygen Species
D017382:Reactive Oxygen Species; D003300:Copper; D048031:JNK Mitogen-Activated Protein Kinases; D003474:Curcumin