Fig 2: Anti-inflammatory effect of corylin on LPS-induced ALI. The experimental results demonstrated that corylin attenuated the overproduction of IL-6 in LPS-activated human bronchial epithelial cells. In intratracheal LPS-induced ALI mice, corylin attenuated tissue damages, suppressed inflammatory cell infiltration, and decreased secretion of IL-6 and TNF-a in the BALF and serum; moreover, it further inhibited the expression of phosphorylation of mitogen-activated protein kinases (MAPKs), including the expression of p-JNK/JNK, p-ERK/ERK, p-p38/p38, and repressed the activation of signal transducer and activator of transcription 3 (STAT3) in lung. Taken together, our results are the first to demonstrate the anti-inflammatory effects of corylin on LPS-induced ALI and suggest corylin has significant potential as a novel therapeutic agent for ALI.

Fig 3: Exogenous overexpression of MG53 inhibited the hepatocarcinogenesis in vivo.To construct xenograft tumor model, 0.1 ml of 107 HUH7 cells were subcutaneously injected to both flanks of the nude mice (4-week-old male BALB/c nude mice, n = 8). When visible tumors appeared, the tumors at the left flanks were injected with Myc-MG53 plasmid while tumors at the right flanks were injected with Myc-vector plasmid. The mice were sacrificed on day 26 after the first injection, and the tumors were separated from both flanks of the mice for further analysis. A, B The tumor growth curve (A) and the image of the isolated tumors from sacrificed mice (B) were presented. C, D The tumor volumes (C) and tumor weights (D) of the indicated groups were analyzed and compared. E qRT-PCR was performed to detect the relative MG53 expression level in MG53 and mock plasmid transfected group. F Protein extracted from the tumors was used for western blot assay to detect the expression levels of Myc, RAC1, p-JNK, p-P38 and p-ERK. G–J Another group of xenograft tumor model was constructed as the above described (4-week-old male BALB/c nude mice, n = 5 for indicated group). When visible tumors appeared, the tumors were divided into three groups including Myc-vector transfected group, Myc-MG53 transfected group and Myc-MG53-?RING transfected group. The tumors were injected with 20 µg of indicated plasmids every other day. The mice were sacrificed on day 28 after the first injection, and the tumors were separated from the nude mice for further analysis. The image of the isolated tumors from sacrificed mice was presented (G). H–J Western blot was performed to detect the successful transfection of the indicated groups (H). Tumor volumes (I) and tumor weights (J) of the indicated groups were analyzed and compared. K The working model showing the role and mechanism of MG53 in HCC cells. MG53 directly binds with RAC1 through its coiled-coil domain and further induces ubiquitous degradation of RAC1 by K48 linked poly-ubiquitination, which leads to inhibition of the RAC1-MAPK pathway; and finally reverses the malignant behaviors of HCC cells and enhances the chemosensitivity of HCC cells to sorafenib. *P < 0.05, **P < 0.01 and ***P < 0.001 for statistical analysis of the indicated groups.

Fig 4: Bioanalysis results of mitogen-activated protein kinase (MAPK) signaling depending on extracellular calcium ions ([Ca2+]e) concentrations of wild-type (WT) CASR and Y825F-mutant CASR (CASR-Y825F) in HEK293 cells. (A) MAPK activity was determined using antibodies against phosphorylated ERK (Thr202/Tyr204), JNK (Thr183/Tyr185), and p38 (Thr180/Tyr182) antibodies. After 10 minutes of calcium treatment, cells were lysed and western blot analysis was conducted. (B) Quantification of phospho-ERK/JNK/p38 based on the expression of total-ERK/JNK/p38 using densitometry analysis (with Image J software). The values were normalized to the expression of wild-type protein in cells not treated with calcium. The level of phospho-ERK/JNK activation was increased in HEK293 cells expressing CASR-Y825F at [Ca2+]e concentrations of 0, 2, 4, and 10 mM. The level of phospho-p38 was increased at a [Ca2+]e concentration of 2 mM. *P<0.05.

Fig 5: Inhibition of ERK-, JNK-, and p38-MAPK signalling pathways block the visfatin-mediated IL-6 expression. (a, b, c, e) Pretreatment of NP cells with ERK1/2, JNK, and p38 inhibitors significantly inhibited the visfatin-mediated induction of IL-6 mRNA and protein expression. (d) ERK, JNK, and p38 phosphorylation rapidly increased after visfatin treatment. ERK, JNK, and p38 activity levels peaked at 30 min post-treatment with visfatin. CTR, control. V, visfatin. n = 3. **P < 0.01 versus contron. ***P < 0.001 versus control. #P < 0.05 versus visfatin. ##P < 0.01 versus visfatin. The data are represented as the mean ± standard deviation