Electroacupuncture Improves Cerebral Ischemic Injury by Enhancing the EPO-JAK2-STAT5 Pathway in Rats

Figures & data

Table 1 Primer Sequences Used for RT-PCR

Gene Names	Primer Names	Sequences	Length
GAPDH	F	AGTGCCAGCCTCGTCTCATA	96
	R	ATGAAGGGGTCGTTGATGGC
HSP70	F	TGCCCTATCCAGATCCTGCT	142
	R	CTCTACTAAGGCCGCACTGG
HIF-1α	F	GCGGCGAGAACGAGAAGAAA	132
	R	GGGGAAGTGGCAACTGATGA
EPO	F	AGAAGATCTGGCCTGGCATC	90
	R	GCAACAGCCATAGCTGGAAGT
JAK2	F	GAGAAGGACCAGACTCCCCT	70
	R	GCACGCACTTCGGTAAGAAC
Bax	F	AAGACAGGGGCCTTTTTGCTA	83
	R	TCCAAGGTCAGCTCAGGTGT
Bcl-2	F	ATGTGTGTGGAGAGCGTCAA	143
	R	GGGCCGTACAGTTCCACAAA
STAT5	F	CACGACGCGAGATTTCTCCA	143
	R	GTGGATGCATTGACGAACCA

Table 2 Neurological Deficit Scores in Each Group (n=12)

Groups	12 h	24 h	48 h	72 h
Sham	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00
Model	2.05±0.34*	2.22±0.36*	2.48±0.37*	2.60±0.21*
EA	2.09±0.49	1.89±0.48	1.36±0.29^#	1.12±0.17^#
AG490-NC	2.11±0.31	2.27±0.32	2.53±0.32	2.70±0.25
AG490	2.47±0.42^#	2.88±0.39^#	3.17±0.32^#	3.75±0.12^#
EA+AG490	2.52±0.37^&	2.72±0.30^&	3.08±0.31^&	3.54±0.15^&@

Notes: Compared to the sham group, *P<0.05; Compared to the model group, ^#P<0.05; Compared to the EA group, ^&P<0.05; Compared to the AG490 group, ^@P<0.05.

Figure 1 Morphologic changes and neuronal apoptosis in the ischemic cortex. (A) Images show HE and TUNEL staining in each group (400×). (B). Statistical analysis of the TUNEL index in each group. The data are presented as the mean ± SD (n = 12). Compared to the sham group, *P<0.05. Compared to the model group, ^#P<0.05. Compared to the EA group, ^&P<0.05. Compared to the AG490 group, ^@P<0.05.

Figure 2 Neuronal ultrastructure in the ischemic cortex at 72 h post-FCI was observed by transmission electron microscopy (uranyl acetate-lead citrate staining, 12, 000×).

Figure 3 EPO-JAK2-STAT5 pathway-related (A) and apoptosis-related (B) gene expression in the ischemic cortex. qRT-PCR was performed to measure the relative mRNA levels of HIF-1α, EPO, JAK2, STAT5, HSP70, Bax and Bcl-2 at 72 h post-FCI. Compared to the sham group, *P<0.05. Compared to the model group, ^#P<0.05. Compared to the AG490-NC group, ^&P<0.05. Compared to the EA group, ^▼P<0.05. Compared to the AG490 group, ^@P<0.05.

Figure 4 EPO-JAK2-STAT5 pathway-related gene and protein expression in the ischemic cortex. (A) Immunohistochemistry was performed to measure the relative protein levels of HIF-1α, EPO, p-JAK2 and p-STAT5 at 72 h post-FCI (400×). (B) MOD of the HIF-1α protein level in each group. (C) MOD of the EPO protein level in each group. (D) MOD of the p-JAK2 protein level in each group. (E) MOD of the p-STAT5 protein level in each group. The data are presented as the mean ± SD (n = 12). Compared to the sham group, *P<0.05. Compared to the model group, ^#P<0.05. Compared to the AG490-NC group, ^&P<0.05. Compared to the EA group, ^▼P<0.05. Compared to the AG490 group, ^@P<0.05.

Figure 5 Apoptosis-related protein expression in the ischemic cortex. (A) Immunohistochemistry was performed to measure the protein levels of HSP70, Bax and Bcl-2 at 72 h post-FCI (400×). (B) MOD of the HSP70 protein level in each group. (C) MOD of the Bax protein level in each group. (D) MOD of the Bcl-2 protein level in each group. The data are presented as the mean ± SD (n = 12). Compared to the sham group, *P<0.05. Compared to the model group, ^#P<0.05. Compared to the AG490-NC group, ^&P<0.05. Compared to the EA group, ^▼P<0.05. Compared to the AG490 group, ^@P<0.05.