Effects of TAN-67 and Naltrindole on striatal expression of p-CREB/CREB at 24 hrs after MCAO.

<p>A, Representative fluorescent micrographs of striatal p-CREB (green) and CREB (red) positive cells in sham and the core of MCAO groups. Bar = 20 µm. B, Representative Western blot images of p-CREB/CREB expression in different groups. C, Quantitative analysis of p-CREB. D, Quantitative analysis of total CREB. N=4. *<i>P</i><0.05 vs. the sham. ^#<i>P</i><0.05 vs. M+TAN67. S, Sham control. M, MCAO. Note that MCAO significantly reduced the expression of CREB and p-CREB. Although DOR inhibition could not further decrease their levels, DOR activation with TAN67 tended to reverse the ischemic reduction.</p

Effect of DOR activation and inhibition on cerebral blood flow.

<p>Administration of aCSF, TAN-67 or Naltrindole had no significant effect on cerebral blood flow in normal rats (<i>P</i>>0.05).</p

Effects of TAN-67 and Naltrindole on striatal expression of BDNF and TrkB at 24 hrs after MCAO.

<p>A, Representative fluorescent micrographs of striatal BDNF positive cells in sham group. Bar = 30 µm. B, Representative Western blot images of BDNF and TrkB expression in different groups. C, Quantitative analysis of BDNF. D, Quantitative analysis of 90 KDa TrkB. E, Quantitative analysis of 140 KDa TrkB. n=4. *<i>P</i><0.05 vs. the sham. ^#<i>P</i><0.05 vs. M+TAN67. S, Sham control. M, MCAO. Note that MCAO significantly reduced the expression of 140 KDa TrkB but not of BDNF and 90 KDa TrkB, while DOR activation with TAN67 largely reversed the ischemic reduction of 140 KDa TrkB expression.</p

Effects of TAN-67 and Naltrindole on cortical expression of pATF-1 and p-CREB/CREB at 24 hrs after MCAO.

<p>A, Representative fluorescent micrographs of cortical p-CREB (green) and CREB (red) positive cells in sham and ischemic penumbra of the MCAO groups. Bar = 20 µm. B, Representative fluorescent micrographs of cortical p-CREB/CREB double-labeled positive cells in ischemic penumbra of the cortex after DOR activation with TAN67. C, Representative fluorescent micrographs of cortical BDNF/CREB double-labeled positive cells in penumbra. Bar = 15 µm. D, Representative Western blot images of pATF-1 and p-CREB/CREB expression levels in different groups. E, Quantitative analysis of pATF-1. F, Quantitative analysis of p-CREB. G, Quantitative analysis of total CREB. n=4. ^*<i>P</i><0.05 vs. the sham. ^#<i>P</i><0.05 vs. M+TAN67. ^&<i>P</i><0.05 vs. the MCAO. S, Sham control. M, MCAO. Note that Naltrindole significantly reduced the expression of pATF-1 with MCAO but not of p-CREB and CREB in the cortex.</p

Effects of TAN-67 and Naltrindole on cortical expression of BDNF and TrkB at 24 hrs after MCAO.

<p>A, Representative fluorescent micrographs of cortical BDNF positive cells in sham group. Bar = 30 µm. B, Representative fluorescent micrographs of cortical BDNF/MAP-2 double-labeled positive cells in sham group. Bar = 15 µm. C, Representative fluorescent micrographs of cortical BDNF/DOR double-labeled positive cells in sham group. Bar = 15 µm. D, Representative Western blot images of BDNF and TrkB expression in different groups. E, Quantitative analysis of BDNF. F, Quantitative analysis of 90 KDa TrkB. G, Quantitative analysis of 140 KDa TrkB. n=4. *P<0.05 vs. the sham. #<i>P</i><0.05 vs. M+TAN67. S, Sham control. M, MCAO. Note that MCAO significantly reduced the expression of 140 KDa TrkB but not of BDNF and 90 KDa TrkB, while DOR activation with TAN67 largely reversed the ischemic reduction of 140 KDa TrkB expression.</p

Effects of TAN-67 and Naltrindole on expression of CD11b at 24 hrs after MCAO.

<p>A, C and E, Representative Western blot images of cortical, striatal and hippocampal CD11b expression in different groups. B, Quantitative analysis of CD11b in the cortex. D, Quantitative analysis of CD11b in the striatum. F, Quantitative analysis of CD11b in the hippocampus. N=4. *<i>P</i><0.05 vs. the sham. ^#<i>P</i><0.05 vs. M+TAN67. S, Sham control. M, MCAO. Note that MCAO significantly increased the expression of CD11b in the cortex, striatum and hippocampus. However, DOR activation specifically attenuated such ischemic increase in the cortex, but not in the striatum and hippocampus.</p

Effects of TAN-67 and Naltrindole on hippocampal expression of BDNF and TrkB at 24 hrs after MCAO.

<p>A, Representative Western blot images of BDNF and TrkB expression in different groups. B, Quantitative analysis of BDNF. C, Quantitative analysis of 90 KDa TrkB. D, Quantitative analysis of 140 KDa TrkB. S, Sham control. M, MCAO. Note that MCAO did not induce any appreciable change in BDNF and TrkB expression in the hipcampus. DOR activation or inhibition also had no significant effect on the expression of these proteins in this region exposed to MCAO.</p

Effects of TAN-67 and Naltrindole on hippocampal expression of pATF-1 and p-CREB/CREB at 24 hrs after MCAO.

<p>A, Representative fluorescent micrographs of hippocampal MAP 2 (green) and CREB (red) positive cells in CA1 in sham group. Bar = 20 µm. B, Representative Western blot images of pATF-1 and p-CREB/CREB expression in different groups. C, Quantitative analysis of pATF-1. D, Quantitative analysis of p-CREB. E, Quantitative analysis of total CREB. N=4. Note that neither MCAO nor DOR activation had any significant effect on the expression of CREB, pCREB and pATF-1 in the hippocampus.</p

Expression of spinal MAPKs after electroacupuncture (EA) treatment in formalin rats.

<p>EA was applied to ST 36 and GB 34 for 30 min before the formalin injection. A shows the representative bands; the density of p-ERK and ERK (B); p-JNK and JNK (C); and p-p38 MAPK and p38 MAPK (D) levels were normalized to β-actin levels and expressed as a fold increase. Data are presented as the mean ± S.E.M. (n = 4 for each group). * p < 0.05, ** p < 0.01.</p

Expression of spinal MAPKs after electroacupuncture (EA) treatment and/or subcutaneous recombinant IL-33 (rIL-33) administration in formalin rats.

<p>EA was applied to ST 36 and GB 34 for 30 min before the formalin injection. Recombinant IL-33 was given subcutaneously immediately before EA treatment. Representative bands (A) and quantification of p-ERK and ERK (B); p-JNK and JNK(C); and p-p38 MAPK and p38 MAPK (D) is shown. Data are presented as the mean ± S.E.M. (n = 4 for each group). * p < 0.05, ** p < 0.01.</p