H e kit

To further confirm VO in the RV, six rats in each group were randomly selected for morphologic examination. Following anesthetization with 5% isoflurane, rat hearts were removed, fixed in 10% paraformaldehyde overnight at room temperature, dehydrated in an ethanol series, embedded in paraffin, and 6-μm-thick sections were prepared. The sections were stained with hematoxylin and eosin (H&E) kits (Beyotime biotech, Shanghai, China) according to the manufacturer's instructions.

After fixing for 24 hours, the right lung was embedded in paraffin, and cut into 4-μm-thick sections, then stained with hematoxylin and eosin (H&E) kit (Beyotime) according to the manufacturer’s instruction.

Recombinant human PDGF was purchased from PeproTech (100-14B, United States). pPERK, ATF6, IRE1α, GRP78, PDI, VPS34, Beclin1, ATG5, LC3II, GAPDH and secondary antibodies were obtained from Abcam (Abcam, United States), PDI was purchased from Cell Signaling Technology (Danvers, MA, United States), NLRP3, Pro-Caspase1, Caspase1, GSDMD, GSDMDP30, IL-1 were obtained from Proteintech (Proteintech, China). Goat anti-rabbit and anti-mouse IgG-HRP were purchased from Cell Signaling Technology (Danvers, MA, United States). An enhanced chemiluminescence (ECL) kit was purchased from Bio-Rad (Hercules, CA, United States). H&E kit was purchased from Beyotime.

Rat kidneys tissues were taken and immersed in 10% formalin solution for 36 h, then were put into an automatic dehydrator (Leica, ASP300S, Munich, Germany) for dehydration, and embedded in paraffin (Leica, Munich, Germany). Paraffin specimen of kidney was cut into 3 μm. After dewaxing with xylene, kidney paraffin sections were immersed in gradient alcohol and water before staining. Hematoxylin stain for 5 min and eosin stain for 10 min were performed, following by dehydration with gradient alcohol and hyalinization with xylene. Then kidney sections were mounted with neutral gum. Finally, histological images were captured using a microscope (Hitachi). The H&E kit was purchased from Beyotime Biotechnology (Cat. No. C0105S, Shanghai, China).

Fetal livers of ADAM17 ^{Zn△/ Zn△}and ADAM17^+/+ embryos were fixed overnight in 4% paraformaldehyde/PBS. Sections of paraffin-embedded tissues were stained using H&E kit (Beyotime, cat No. C0105, Haimen, China) followed by analysis and quantification. In brief, a total of eight mice from each strain were used, and over five sections from a similar location of each fetal liver were obtained. Ten random views were selected for microscopic examination and quantification for each section.

Gelatin type B, tragacanth gum, Curcumin, Glacial acetic acid, Sodium sulfate, 37% formaldehyde, Chondroitin sulfate, EDC, NHS were purchased from Soleberg Reagent. Collagen (Type II, Chicken, Protein 60%, Yuanye), Fuchsin Complete Adjuvant (FCA), Fuchsin Incomplete Adjuvant (FICA) was purchased from Genye Reagent Company. Nitrendipine was obtained from Aladdin Chemical Reagent Co. CCK-8 kit, ROS kit, malondialdehyde (MDA) content assay kit, total glutathione (GSH-Px) content assay kit, superoxide dismutase (SOD) content assay kit and H&E kit were purchased from Beyotime Anti-HIF-1α, Anti-iNOS, anti-Arg-1, anti-IL-1β, anti-IL-10 were purchased from Abcam. RAW264.7 cells were obtained from Chinese Academy of Sciences.

After 6 weeks, animals were anaesthetized with isoflurane and euthanized by cardiac puncture. Mouse kidney was isolated after perfusion with cold salt solution at a pressure of 100 mmHg. The kidney tissues were fixed in the 10% formaldehyde, dehydrated, immersed in wax, embedded and sliced into sections (2–4 µm).
Histopathological staining of kidney sections was performed with hematoxylin–eosin (H&E) kit (C0105, Beyotime Institute of Biotechnology, Shanghai, China), Masson’s trichrome stain kit (1004850001, Sigma), and periodic acid–Schiff (PAS) reagent (ab150680, Abcam, Cambridge, UK) according to the manufacturer’s instructions. After sealing with neutral gum, the sections were observed and photographed under an inverted microscope (IX73, Olympus, Tokyo, Japan). All morphological analyses were performed by two individual experienced pathologists in a double-blind manner.