Anti mmp3

Incomplete Freund's adjuvant (IFA) and Bacillus Calmette-Guérin (BCG) were purchased from Rebio Scientific (Shanghai, China). TNF-α, IL-1β, IL-10, and IL-6 ELISA kits were the products of Multi Science (Hangzhou, China). Biochemical quantification kits for the determination of malondialdehyde (MDA), reduced glutathione (GSH), total superoxide dismutase activity (SOD), total antioxidant capacity (T-AOC), nitric oxide (NO), nitric oxide synthase (inducer) (iNOS), and carbonic oxide (CO) were brought from Jiancheng Bioengineering Institute (Jiangsu, China). The primary antibodies used in immunoblotting and immunohistochemical experiments including anti-MMP3, COX-2, TLR4, SIRT1, PPAR-γ, p65, p-p65, and β-actin antibodies together with secondary antibodies were provided by ABclonal Technology (Wuhan, China). Fluorescein-tagged anti-CD3, CD4, and IL-17α antibodies for the use of flow cytometry analysis were supplied by Beyotime Biotech (Nantong, Jiangsu, China). Rat peripheral blood mononuclear cell (PBMC) isolation kit, TaqMan RT-qPCR kit, and cDNA synthesis kit were purchased from Solarbio Technologies (Beijing, China). All the solvents were of analytical grade and supplied by Merck Chemicals (Shanghai, China). High-purity compounds (>98%) kaempferol, paeoniflorin, matrine, sophocarpine, and sinomenine were all obtained from Yuanye Bio-Technology (Shanghai, China).

TAPI-1, L-685,458 and batimastat were purchased from Selleck. BACE inhibitor IV (BSI IV) was purchased from Calbiochem. Cell Titer-Glo was from Promega. The western blotting assays were achieved with the following antibodies: anti-actin (A2066, Sigma), anti-ADAM10 (Ab1997, Abcam), anti-BACE1 N-terminus (AP7774b, Abgent), anti-APP-CTF (A8717, Sigma), anti-Nicastrin(NCT) (N1660, sigma), anti-PS1 N terminal (Covance), anti-Pen2 (P5622, Sigma) anti-sAPPα (IBL), anti-human sAPPβ-wild type (IBL), anti-MMP3 (A6260, ABclonal), anti-MMP9 (A0289, ABclonal).

Human and mouse aortas or cultured SMCs were harvested and lysed using RIPA Lysis Buffer (Beyotime Biotechnology). Equal amount (15‐30 μg) of total protein was fractionated on 5%‐15% SDS‐polyacrylamide gel and transferred onto a PVDF membrane (Thermo Fisher Scientific, Waltham, MA, USA). Then the blots were incubated with primary antibodies: anti‐GDF11 (1:1000, abcam, Cambridge, MA, USA), anti‐ACTA2 (1:1000, Novus Biologicals), anti‐MMP‐2 (1:500, Proteintech, Rosemont, IL, USA), anti‐MMP‐9 (1:1000, Proteintech), anti‐MMP‐3, anti‐TNF‐α (1:500, ABclonal, Wuhan, China), anti‐IL‐6, anti‐p‐Smad‐2, and anti‐p‐Smad‐3 (1:1000, ABclonal) at 4°C overnight. The membranes were probed with species‐relevant HRP‐linked secondary antibodies (1:10 000, Proteintech) at 37°C for 40 minutes. Besides, housekeeping protein β‐actin (1:2000, Proteintech) was used as the internal control.