Rats were administered intraperitoneally with pirfenidone (250 mg/kg) (Selleck Chemicals, Houston, TX, United States) (Shihab et al., 2002 (link); Burghardt et al., 2007 (link)), SIS3 HCl (2.5 mg/kg) (Selleck Chemicals, Houston, TX, United States) (Li et al., 2010 (link); Liu et al., 2018 (link)) and cyclopamine (10 mg/kg) (Cayman Chemical, Ann Arbor, MI, United States) (Alvarez et al., 2011 (link); Yu et al., 2017 (link)) 30 min before ischemia.
Rats were randomly divided into 9 groups: animals received sham operation and an equal volume of DMSO (Sham); rats received MCAO/R and an equal volume of DMSO (I/R); MCAO/R rats were treated with TGF-β2 inhibitor pirfenidone or Smo inhibitor cyclopamine (I/R + Pir, I/R + CYC); MCAO/R rats were treated with 1.5% ISO post-conditioning for 1 h after immediate reperfusion (ISO); MCAO/R rats were treated with pirfenidone, Smad3 inhibitor SIS3 HCl, cyclopamine, pirfenidone combined with cyclopamine before ISO post-conditioning (ISO + Pir, ISO + SIS3, ISO + CYC, and ISO + Pir + CYC, respectively).
Rats were randomly divided into 9 groups: animals received sham operation and an equal volume of DMSO (Sham); rats received MCAO/R and an equal volume of DMSO (I/R); MCAO/R rats were treated with TGF-β2 inhibitor pirfenidone or Smo inhibitor cyclopamine (I/R + Pir, I/R + CYC); MCAO/R rats were treated with 1.5% ISO post-conditioning for 1 h after immediate reperfusion (ISO); MCAO/R rats were treated with pirfenidone, Smad3 inhibitor SIS3 HCl, cyclopamine, pirfenidone combined with cyclopamine before ISO post-conditioning (ISO + Pir, ISO + SIS3, ISO + CYC, and ISO + Pir + CYC, respectively).
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