Figure 4.

Methanol extract from Impatiens balsamina (MEIB) disrupts mitochondrial membrane potential by regulating the expression of Bcl-2 family proteins. HSC-2 cells were treated with dimethyl sulfoxide (DMSO) or 60 μg/ml of MEIB for 24 h. (a) The whole-cell lysates were analyzed by Western blot analysis using antibodies against t-Bid, Bak, Bad, Bax, Puma, Bim, Bcl-xL, Mcl-1 and Bcl-2. *P < 0.05 compared with the DMSO-treated group; (b) Mitochondrial membrane potential was assessed using a 5,5’,6,6’-tetrachloro-1,1’,3,3’-tetraethylbenzimidazolyl carbocyanine iodide assay. The fluorescence intensity of Red/Green expressed as the means ± standard deviation of triplicate experiments. *P < 0.05 compared with the DMSO-treated group; (c) detection of cytochrome C in cytosolic fractions by Western blot analysis. Cox4 and α-tubulin were used as a fraction marker for mitochondrial and cytosolic fraction, respectively. *P < 0.05 compared with the DMSO-treated group, (d) cleaved caspase-9 and cleaved caspase-3 were detected by Western blot analysis