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

Figure 7

E2 alleviates the inhibition of miR-29a-5p induced by LPS. (a, b) RAW 264.7 cells were treated with the indicated concentrations of LPS for 24 h. qRT-PCR detected the expression of miR-29a-5p. Relative expression of miR-29a-5p was presented after normalizing to U6 (; ). and vs. the LPS 0 μg/ml group, respectively. (b) RAW 264.7 cells were treated with 1 μg/ml LPS for 0 h, 6 h, 12 h, and 24 h. The expression of miR-29a-5p was analyzed by qRT-PCR. Relative expression of miR-29a-5p was presented after normalizing to U6 (; ). and vs. the 0 h group, respectively. (c) 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 detected the expression of miR-29a-5p. Relative expression of miR-29a-5p was presented after normalizing to U6 (; ). vs. the E2 0 nM group. (d) RAW 264.7 cells were cultured in a medium containing 1 μg/ml LPS plus 100 nM E2 for the indicated times. qRT-PCR detected the expression of miR-29a-5p. Relative expression of miR-29a-5p was presented after normalizing to U6 (; ). and vs. the 0 h group, respectively. (e) 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 miR-29a-5p was detected by qRT-PCR. Relative expression of miR-29a-5p was presented after normalizing to U6 (; ). vs. the LPS group. ^### vs. the LPS+E2 group. (f) qRT-PCR analysis of miR-29a-5p expression in NC-, CLP-, and CLP+E2-treated vascular tissues. Relative expression of miR-29a-5p was presented after normalizing to U6. in each group. vs. the NC group. ^### vs. the CLP group.

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