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Table 2 Effects of sema 3A on autonomic abnormalities 5 weeks after infarction

From: Semaphorin 3A attenuates cardiac autonomic disorders and reduces inducible ventricular arrhythmias in rats with experimental myocardial infarction

   Sham MI-CON MI-GFP MI-SiRNA MI-Sema MI-PBS
1-RT-PCR (target mRNA /GAPDH) TH 1 2.93 ± 0.21 2.87 ± 0.18 3.82 ± 0.34* 1.84 ± 0.14** 3.12 ± 0.23***
CHAT 1 1.66 ± 0.14 1.71 ± 0.17 1.67 ± 0.16 1.61 ± 0.14**** 1.72 ± 0.17
2- IHC (um2/mm2) TH-positive 1281 ± 75.1 3589 ± 278.1 3539 ± 176 4707 ± 193.2* 2330 ± 162.4** 3564 ± 118.9***
  Sema 3A protein 0.14 ± 0.02 0.35 ± 0.03 0.33 ± 0.025 0.167 ± 0.02* 0.44 ± 0.03** 0.31 ± 0.02***
3- WB (target protein/GAPDH) TH protein 0.19 ± 0.03 0.45 ± 0.035 0.47 ± 0.029 0.67 ± 0.027* 0.35 ± 0.014** 0.48 ± 0.03***
CHAT protein 0.085 ± 0.087 0.174 ± 0.013 0.174 ± 0.01 0.173 ± 0.011 0.167 ± 0.011** 0.179 ± 0.09
TH/CHAT 2.18 ± 0.25 2.63 ± 0.26 2.67 ± 0.19 3.77 ± 0.20* 2.66 ± 0.18** 2.10 ± 0.10***
4- HPLC (pmol/mg) NE 2.1 ± 0.12 4.3 ± 0.29 4.1 ± 0.31 5.1 ± 0.27* 2.9 ± 0.26** 3.9 ± 0.27***
ACh 2.9 ± 0.26 3.9 ± 0.23 3.8 ± 0.27 3.5 ± 0.24 3.6 ± 0.28**** 3.8 ± 0.27
  1. (1) RT-PCR was used to assess the mRNA expression of TH, and CHAT in various groups. Sema 3A silencing increased the expression of GAP 43 and TH compared with the MI-GFP group (P < 0.05). However, the administration of sema 3A reduced the expression in TH compared with the MI-PBS group (P < 0.05). CHAT mRNA expression was comparable between various MI groups (P > 0.05). (2) Immunohistochemistry results revealed that the density of TH- and GAP 43- positive nerve fibers were higher in sema 3A-silenced infarcted-hearts compared with the MI-GFP group (both P < 0.05). Recombinant sema 3A reduced the number of TH- and GAP 43- positive nerve fibers compared with the MI-PBS group (both P < 0.01). There was no difference in the density of TH- nerve fibers among the MI-CON, MI-GFP, and MI-PBS groups (all P > 0.05). (3) Western blot (WB) showing increased protein expression of TH and CHAT in the MI-CON group compared with the sham group (P < 0.05). Sema 3A silencing increased the expression of TH protein, whereas sema 3A administration reduced TH protein expression. CHAT protein expression was comparable among the various MI groups. However, the ratio of TH/CHAT protein expression was increased in the MI-SiRNA group compared with the MI-GFP group (P < 0.05). The ratio was lower in the MI-Sema group compared with the MI-PBS group (P = 0.021), but there was no significant difference between the MI-Sema and sham groups (P = 0.34). (4) HPLC was used to assess local sympathetic and parasympathetic activity. MI resulted in increased myocardial NE and Ach levels in the MI-CON group compared with the sham group (both P < 0.05). NE content was increased in the MI-SiRNA group compared with the MI-GFP group (P = 0.007). In addition, NE content was reduced in the MI-Sema group compared with the MI-PBS group (P = 0.024). However, myocardial Ach levels were similar in the various infarcted groups (P > 0.05). * P < 0.05 vs. MI-GFP group, ** P < 0.05 vs. MI-SiRNA group, *** P < 0.05 vs. MI-sema group, **** P > 0.05 among various MI groups