Figure 8.

Enhanced regeneration induced by SG is independent of the Yap function. (A) The correlation analysis heatmap of serum from SG and Sham groups of mice showed high similarity in serum composition, but differences existed between SG and Sham groups. (B) A total of 79 serum proteins were detected, and the Venn diagram showed that there were three differentially expressed proteins in SG compared to the Sham group. (C) Target protein screening was conducted, and the target protein needed to meet the following criteria: closely related to classical liver regeneration signals IL6 and Tnf, more closely related to the three differentially expressed proteins detected above, and able to mediate tumor occurrence. The result showed that Yes1 was the most suitable candidate based on the screening criteria. (D) Western blot was used to detect the Hippo–Yap signal in SG versus Sham groups, and the results showed that the Yap signal in the mouse liver was strongly inhibited 6 weeks after SG. (E) In order to evaluate the effect of SG on the Yap signal throughout the postoperative period, Yap immunofluorescence staining was performed on mice that underwent combined surgery 7 days after the operation, and the results showed that SG seemed to enhance Yap function in the early postoperative period. (F) In order to verify whether Yap is a necessary factor for SG to produce liver regeneration enhancement and metabolic improvement during the long-term postoperative process, verteporfin was used to inhibit Yap function in mice, and surgical intervention and a control group were set up simultaneously. (G) The weight loss effect induced by SG was not influenced by verteporfin treatment. (H) The Western blot results demonstrated that verteporfin effectively suppressed the expression levels of YAP. (I) Gross observation of the liver showed that compared to the Sham+VP group, the other two groups had significant and almost similar relief of steatosis. (J) Serological test results indicate that verteporfin did not affect the beneficial effects of liver function improvement induced by SG. (K) The results of BODIPY staining showed that Yap inhibition did not affect the ability of SG to relieve fatty liver, and (L) the results of BrdU staining showed that the enhancement of liver regeneration produced by SG was not affected by Yap inhibition. (Data are shown as mean±SE. ns=P>0.05, **=P<0.01, ***=P<0.001, and ****=P<0.0001. For panels B and D, nine visual fields were collected to calculate the BrdU-positive rate. For panel G, n=6 Sham+VP, 8 SG, 7 SG+VP. For panel J, n=5 Sham+VP, 5 SG, 5 SG+VP). SG, sleeve gastrectomy.