Alzet mini osmotic pumps model 1002

After 1 week of acclimatization, two mice were placed into each cage, to avoid social isolation, and cage-mates were not changed during the experiment. Chronic TGF-β2 infusion was performed as previously described (Takahashi et al., 2019 (link)). Briefly, the animals were randomly allocated to three groups at 10 weeks of age: WT + vehicle; db/db + vehicle, and db/db + TGF-β2 groups. Under isoflurane anesthesia (induction: 4%, maintenance: 2%), the mice underwent surgery to implant Alzet mini-osmotic pumps (model 1002, Durect, Cupertino, CA, United States) subcutaneously. The osmotic pumps were filled with recombinant TGF-β2 (8406LC, Cell Signaling Technologies, Danvers, MA, United States) diluted in PBS, or PBS alone for the WT + vehicle and db/db + vehicle groups. For 12 consecutive days, the mice received 12 ng TGF-β2 each per hour. After euthanasia, the pumps were removed and the contents were confirmed to have been completely discharged. The dose and duration of treatment were those used in a previous study (Takahashi et al., 2019 (link)).

Male 8 to 10-week-old male C57/B6 mice were purchased from the Shanghai Experimental Animal Center, China. The investigation conforms with the Guide for the Care and Use of Laboratory Animals published by the United States National Institutes of Health (NIH Publication No. 85-23, revised 1996) and was approved by the Ethics Review Board of Ruijin Hospital, Shanghai, China. Four groups of animals were included: vehicle infusion (0.9% NaCl solution, control), vehicle infusion+EMD638683, AngII infusion, Ang II infusion+EMD638683. Ang II (1000 ng/kg/min) was infused for 2 weeks using Alzet mini-osmotic pumps (model 1002, DURECT, Cupertino, CA, United States) implanted subcutaneously in the surgery. EMD638383 was delivered i.p daily at a dosage of 10 mg/kg body weight. Systolic blood pressure was measured on every week by tail-cuff plethysmography (Visitech Systems, Apex, NC, United States) as a control for efficient AngII infusion. All surgical procedures were practiced under anesthesia.