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. 2016 Nov 10;6:36530. doi: 10.1038/srep36530

Figure 5. Western blot analysis in the in vitro study.

Figure 5

In the in vitro study, we treated cells with 20 μM Hemin and (or) 10 μM ZnPP for 1 h prior to LPS treatment and continued the incubation for 24 h. As illustrated in (A–D), HO-1 expression was upregulated in the cells with LPS and pretreatment, compared with that of the unstimulated cells in the control group(C) (P < 0.05; (A). To investigate whether HO-1 impacts Mfn1 in LPS-induced rat alveolar macrophages, we analyzed the expression of the mitochondrial fusion protein Mfn1. As our results indicated, LPS significantly decreased Mfn1 expression compared with that of the control group(C) (P < 0.05); however, after Hemin pretreatment, Mfn1 expression was upregulated compared with that of the LPS group(L) (P < 0.05; (B). In addition, it has been reported that HO-1 is regulated by the PI3K/Akt pathway, and we thus investigated PI3K/Akt expression in LPS-induced rat alveolar macrophages. As our data showed, similar to HO-1 expression, PI3K/Akt expression was remarkably enhanced in the LPS group(L) compared with that of the control group(C) (P < 0.05; (C,D). Moreover, in the LPS + Hemin group(LH), the level of PI3K/Akt expression was upregulated to a higher degree compared with that of the LPS group(L) (P < 0.05; (C,D). By contrast, after ZnPP-IX pretreatment, our data showed a lower level of PI3K/Akt expression in the LPS + ZnPP-IX group(LZ) compared to that of the LPS group(L) (P < 0.05; (C,D).