Evidence Details for Ang2
PMID Title Journal Year Abstract
35321503 Scalp Electroacupuncture Promotes Angiogenesis after Stroke in Rats by Activation of Wnt/beta-Catenin Signal Pathway. Evid Based Complement Alternat Med. 2022 Mar 14;2022:1649605. doi: 10.1155/2022/1649605. eCollection 2022. 2022 BACKGROUND: Scalp acupuncture is a contemporary acupuncture method based on the fundamental theories of traditional acupuncture, which has been widely used in patients with stroke in China. However, the effectiveness is controversial due to lack of solid experimental evidence. METHODS: In this study, a rat model of cerebral ischemia-reperfusion injury (CIRI) was established by the middle cerebral artery occlusion/recirculation. The efficacy of scalp acupuncture against CIRI was evaluated by the mNSS scores, TTC staining for brain slices, and laser Doppler perfusion imaging. Immunohistochemical staining for angiogenetic factors indicated the vascularization after CIRI, including VEGF, Ang2, and bFGF. Activation of the Wnt/beta-catenin signaling pathway and p-GSK3beta (ser9)/VEGF pathway in the injured brain tissues was assessed by western blotting and qRT-PCR. RESULTS: On the 7, 14, and 21 days after CIRI, scalp acupuncture could reduce the mNSS scores, decrease the cerebral infarction area, and accelerate the recirculation of ischemic brain tissues. VEGF, FLK1, bFGF, and Ang2 were upregulated on both the mRNA and protein levels in the ischemic brain tissues of the AC group, suggesting that the recirculation might result from angiogenesis, which was also confirmed with the IHC staining in the angiogenetic markers of VEGF, Ang2, and bFGF. Moreover, Wnt3a, beta-catenin, and cyclin D1 were also upregulated on both the mRNA and protein levels in the ischemic brain tissues of the AC group on day 7, 14, and 21, indicating that the Wnt/beta-catenin signaling pathway was activated after the treatment of scalp acupuncture. In contrast, dikkoppf-1 (DKK1) pretreatment, a specific inhibitor for the Wnt/beta-catenin signaling pathway, inactivated the Wnt3a/beta-catenin signaling pathway and exacerbated the infarct size induced by the cerebral IR injury on day 7. CONCLUSION: Together, our findings demonstrated a mechanism whereby scalp acupuncture led to the activation of Wnt/beta-catenin signaling pathway, promoting angiogenetic factor expression and restoring blood perfusion in the ischemic zone."

Evidence Sentence: Immunohistochemical staining for angiogenetic factors indicated the vascularization after CIRI, including VEGF, Ang2, and bFGF.
Evidence Sentence: VEGF, FLK1, bFGF, and Ang2 were upregulated on both the mRNA and protein levels in the ischemic brain tissues of the AC group, suggesting that the recirculation might result from angiogenesis, which was also confirmed with the IHC staining in the angiogenetic markers of VEGF, Ang2, and bFGF.
Evidence Sentence: Immunohistochemical (IHC) Staining of VEGF, Ang2, and bFGF
Evidence Sentence: In an IHC assay, proangiogenic factors such as VEGF, Ang2, and bFGF were labeled and observed in the brain tissue sections at each time points (Figure 6).
Evidence Sentence: Compared with the sham group, the positive staining of VEGF, Ang2, and bFGF showed a compensatory increase in the model group and a significant increase in the brain tissue from the AC group reaching maximum expression on day 21.
Evidence Sentence: VEGF, bFGF, and Ang2 expression levels were elevated in the model group from brain tissues compared with those from the sham group (P < 0.05) on days 14 and 21.
Evidence Sentence: However, VEGF, FLK1, bFGF, and Ang2 expression levels were not different between the model and sham groups on day 7.
Evidence Sentence: The treatment of acupuncture showed increased expression of VEGF, FLK1, bFGF, and Ang2 compared with the model group on days 7, 14, and 21 (P < 0.05 or P < 0.01).
Evidence Sentence: To validate the expression level of the proangiogenic proteins after CIRI, we also detected the expression levels of VEGF and Ang2 after treating rats with DKK1.
Evidence Sentence: As shown in Figure 2(c), on day 7, compared with the model group, the expression levels of VEGF and Ang2 deceased in the brain tissues from the group of DKK1 (P < 0.05 or P < 0.01).
Evidence Sentence: VEGF and Ang2 expression levels were increased in the AC + DKK1 group compared with the DKK1 group (P < 0.01) on day 7.
Evidence Sentence: Compared with the AC + DKK1 group, VEGF and Ang2 expression levels were increased in the AC group (P < 0.05) on day 7.
Evidence Sentence: Moreover, compared with the model group, both bFGF and Ang2 mRNA levels increased on days 14 and 21 (P < 0.05 or P < 0.01).
Evidence Sentence: Similar to the protein assay, we also detected mRNA levels of VEGF and Ang2 by RT-PCR after adding DKK1.
Evidence Sentence: As shown in Figure 2(e), on day 7, mRNA levels of VEGF and Ang2 were reduced in the DKK1 group compared with the model group (P < 0.05 or P < 0.01).
Evidence Sentence: VEGF and Ang2 mRNA levels were increased in the AC + DKK1 group compared with the DKK1 group (P < 0.01) on day 7.
Evidence Sentence: Compared with the AC + DKK1 group, VEGF and Ang2 expression levels increased in the AC group (P < 0.05) on day 7.