Extracellular HMGB1 Induced Glomerular Endothelial Cell Injury via TLR4/MyD88 Signaling Pathway in Lupus Nephritis

Figure 6

Inhibition of the TLR4/MyD88/NF-κB/p65 pathway relieved GEC injury in HMGB1-induced LN. (a) Western blotting showed the expression of TLR4, NF-κB/p65, p-IκBα, and MyD88 in HRGECs cultured with LN plasma. (b) IF staining for NF-κB/p65 (red) in HRGECs cultured with LN plasma. (c, e, g, and h) Western blotting of TLR4, MyD88, and p-IκBα in LN plasma-stimulated HRGECs pretreated with different inhibitors. (d, f, and i) IF staining of NF-κB/p65 (red) in LN plasma-stimulated HRGECs pretreated with different inhibitors. (j, k) TEER and permeation in LN plasma-stimulated HRGECs pretreated with different inhibitors (, ). (l) The level of SDC-1 in supernatant of LN plasma-stimulated HRGECs pretreated with different inhibitors measured by ELISA. (m) IF staining of SDC-1 in LN plasma-stimulated HRGECs pretreated with different inhibitors. (n) IF staining of VE-cadherin (green) and F-actin (red) in LN plasma-stimulated HRGECs pretreated with different inhibitors. (o–q) Western blotting of VCAM-1 in LN plasma-stimulated HRGECs pretreated with different inhibitors. (r) The level of NO in LN plasma-stimulated HRGECs pretreated with different inhibitors. vs. control group; ^# vs. LN group.

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