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Souza et al. Neuroimmunol Neuroinflammation 2019;6:12 I http://dx.doi.org/10.20517/2347-8659.2019.04 Page 7 of 13

A B
BV
Sham CTL BV (NP) (ST36+GV3)

Figure 3. Influence of apipuncture on IBA-1 levels in the spinal cord 1, 3 and 5 days after SCI. In A: representative of the bands of proteins
of IBA-1 by the groups Sham, CTL-SCI, BV (NP)-SCI and BV (ST36 + GV3)-SCI measured by densitometry, normalized by β-actin; In B: the
comparison of the IBA-1 protein content at the spinal cord lesion site of rats submitted to SCI and apipuncture at ST36 and GV3 points [BV
(ST36)-SCI; n = 6], SCI and apipuncture at non-acupoints [BV (NP)-SCI, n = 6], only SCI (CTL-SCI, n = 6) and Sham-SCI (n = 6). Values
are presented as mean ± SEM. ***P < 0.001 compared to CTL-SCI and BV (NP)-SCI. BV: bee venom; NP: non-acupoints; SCI: spinal cord
injury

A B
BV
Sham CTL BV (NP) (ST36+GV3)

Figure 4. Influence of apipuncture on BCL levels in the spinal cord 1, 3 and 5 days after SCI. In A: representative of the bands of proteins of
BCL-2 by the groups Sham, CTL-SCI, BV (NP)-SCI and BV (ST36 + GV3)-SCI measured by densitometry, normalized by β-actin; In B: the
comparison of the BCL-2 protein content at the spinal cord lesion site of rats submitted to SCI and apipuncture at ST36 and GV3 points [BV
(ST36)-SCI; n = 6], SCI and apipuncture at non-acupoints [BV (NP)-SCI, n = 6], only SCI (CTL-SCI, n = 6) and Sham-SCI (n = 6). Values
are presented as mean ± SEM. ***P < 0.001 compared to CTL-SCI and BV (NP)-SCI. BV: bee venom; NP: non-acupoints; SCI: spinal cord
injury

Apipuncture reduces activation of microglia/macrophages
The group BV (ST36 + GV3)-SCI showed significant lower levels of IBA-1 (microglia/macrophage active
marker) protein compared to CTL-SCI and BV (NP)-SCI groups at all times evaluated (1st, 3rd and 5th
days) after SCI (One way ANOVA followed by Bonferroni test; P < 0.001) [Figure 3].

Apipuncture increases anti-apoptotic factor BCL-2 and neuroprotection of neurons and
oligodendrocytes
The group BV (ST36 + GV3)-SCI showed significant higher levels of BCL-2 (anti-apoptotic factor marker),
compared to CTL-SCI and BV (NP)-SCI groups on the 5th day after SCI (One way ANOVA followed by
Bonferroni test; P < 0.001) [Figure 4]. Furthermore the group BV (ST36 + GV3)-SCI showed significant
higher levels of NeuN (neuron marker) and CNPase protein (enzyme marker expressed by viable
oligodendrocytes), compared to CTL-SCI and BV (NP)-SCI groups on the 7th day after SCI (One way
ANOVA followed by Bonferroni test; P < 0.001) [Figure 5].