Immunohistochemistry
After being baked at 60 °C for 2 hours, the paraffin sections were dewaxed and hydrated in turn, washed with PBS and incubated with 3% hydrogen peroxide for 5-10 minutes (at room temperature) to eliminate the activity of endogenous peroxidase. After antigen repair, goat serum was added and incubated at room temperature for 15 minutes. The primary antibody working solution containing TRPV1 (1:200), Kir6.1 (1:200), SUR2B (1:200), and eNOS (1:200) was added to cover the tissue, and the tissue was incubated at 4 °C overnight. After being rinsed, the secondary antibody was diluted with PBS, added and incubated at room temperature for 2 hours. Then, DBA reagent was used for color development, the samples were stained again with hematoxylin and subjected hydrochloric acid alcohol differentiation, dehydration, transparency, and sealing with neutral gum. Then, the images were observed and photographed under a microscope. The expression of various proteins in endothelial cells was mainly observed. Brown–yellow staining indicated positive expression, and the relative expression level is represented by the optical density value. The images were analyzed by Q-IMAGING software.