| ObjectiveTo study the effect of zinc on expression of ZnT-7 in proliferation of grow th plate chondrocytes and its regularity in rat.MethodsGrowth plate chondrocytes cell were isolated from rib cartilage of 60-to 8 0-g wista rat. Low zinc cell model was established by TPEN, a kind of chelati ng agent which chelates specially to zinc. The rate of the the necrotic cells an d the secondary necrotic cells were analyzed by flow cytometry (Annexin V-FIT C/PI) Localization of the ZNT-7 protein was used Fluorescent staining. Real-time quantitative PCR and the Western-Blot method was used to measure expression levels of ZNT-7 the gene and the proteinum expression in zinc depleted Growt h Plate Chondrocytes cell on various concentration of TPEN exposure(0,5,10 and 20μmol/L 12h)ResultThe necrotic cells and the secondary necrotic cells rate of the Growth plate chondrocytes cells were significantly increased when exposed to TPEN. The results show that the necrotic cells and the secondary necrotic cells rate reached 2.25%and 2.24%in 0 group,3.06%and 8.78%in 5umol/L group.5.15%and 16.89%In 5μmol/L group.10.03%and 17.21%in 5μmol/L group.The result of the immunofluorescence show that ZnT5 was found in the cytoplasmic vesicles and perinuclear portion in Growth Plate Chondrocytes cells. The immunostaining against marker proteins, then, reveal the specific marker (Golgi) that was completely superimposed with ZnT5. Zinc depletion induced by the zinc chelator TPEN (N,N,N',N'-Tetrakis-(2-pyridylmethyl) ethylene diamine) resulted in a up to induction of transporters ZnT-7 mRNA levels.The result of the Western Blot show the same trend. The experiment was repeated three, that use spssll.O,software, t test analysis of variance of variance test data to p<0.05 for the difference was significant.ConclusionZinc can regulate the expression of Znt-7 in Growth Plate Chondrocytes cells, Simultaneously, Zinc deficiency can increase the call early apoptosis and late apoptosis. |
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