17β-Estradiol Attenuates LPS-Induced Macrophage Inflammation In Vitro and Sepsis-Induced Vascular Inflammation In Vivo by Upregulating miR-29a-5p Expression

Figure 3

E2 inhibits LPS-induced macrophage inflammation by downregulating NLRP3. (a, b) RAW 264.7 cells were cultured in a medium containing 1 μg/ml LPS and treated with different doses of E2 for 24 h. qRT-PCR (a) and Western blotting (b) detected the mRNA and protein expression of IL-1β and NLRP3. Relative expression of IL-1β and NLRP3 mRNA was presented after normalizing to GAPDH (; ). and vs. the E2 0 nM group, respectively. (c, d) RAW 264.7 cells were treated with 1 μg/ml LPS and 100 nM E2 for different times. The expression of IL-1β and NLRP3 mRNA and protein was analyzed by qRT-PCR (c) and Western blotting (d). Relative expression of IL-1β and NLRP3 mRNA was presented after normalizing to GAPDH (; ). , , and vs. the 0 h group, respectively. (e, f) RAW 264.7 cells were treated with LPS (1 μg/ml), LPS plus E2 (100 nM), or LPS plus E2 and ER antagonist ICI182780 (1 μM). The expression of IL-1β and NLRP3 was analyzed and presented as in (a, b). Relative expression of IL-1β and NLRP3 mRNA was presented after normalizing to GAPDH (; ). vs. the LPS group. ^### vs. the LPS+E2 group. (g, h) RAW 264.7 cells were infected with pcDNA 3.1 or pcDNA 3.1-NLRP3 for 24 h, then incubated with 1 μg/ml LPS and 100 nM E2 for 24 h. The expression of NLRP3 and IL-1β mRNA and protein was analyzed and presented as in (a, b). Relative expression of NLRP3 and IL-1β mRNA was presented after normalizing to GAPDH (; ). vs. the pcDNA 3.1 group. ^# vs. the pcDNA 3.1-NLRP3 group.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)