Cross electro-nape-acupuncture ameliorates cerebral hemorrhage-induced brain damage by inhibiting necroptosis

Figure 1 Cerebral hemorrhage induced necroptosis in brain tissue. A and B: Propidium iodide staining (A) and propidium iodide positive rate (B) in brain tissues of the experimental rats; C: Immunofluorescence assay showed the expression of the necroptotic marker p-MLKL in the experimental rats. Scale bar: 20 μm. ^cP < 0.001. CH: cerebral hemorrhage; PI: Propidium iodide; MLKL: mixed lineage kinase domain like pseudokinase.

Figure 2 Cross electro-nape acupuncture alleviated brain damage in rats with cerebral hemorrhage. A: Neurological scores in the experimental rats; B: Behavioral scores in the experimental rats; C: Evaluation of cerebral edema in the brain tissues of experimental rats. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. CENA: Cross electro-nape acupuncture; CH: cerebral hemorrhage.

Figure 3 Cross electro-nape acupuncture inhibited cerebral hemorrhage -mediated necrosome formation in vivo. A and B: Propidium iodide staining (A) and propidium iodide positive rate (B) in the brain tissues of experimental rats; C: Protein level of RIPK1, RIPK3, p-RIPK3, MLKL and p-MLKL in the brain tissues of experimental rats; D-H: Quantitative analysis of RIPK1 (D), RIPK3 (E), p-RIPK3 (F), MLKL (G) and p-MLKL (H); I-K: RIPK1 and RIPK3 interaction in the brain tissues of experimental rats using IP assay (J) and quantitative analysis (K) of the IP assay. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. CENA: Cross electro-nape acupuncture; CH: cerebral hemorrhage; RIPK: Receptor interacting protein kinase; MLKL: mixed lineage kinase domain like pseudokinase.

Figure 4 Cross electro-nape acupuncture ameliorated inflammatory cytokines in rats with cerebral hemorrhage. A-C: The mRNA level of IL-6 (A), IL-8 (B) and TNF-α (C) in the brain tissues of experimental rats; D-F: The concentration of IL-6 (D), IL-8 (E) and TNF-α (F) in the CSF of rats; G-I: The secretion level of IL-6 (G), IL-8 (H) and TNF-α (I) in the serum of rats. ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. CENA: Cross electro-nape acupuncture; CH: cerebral hemorrhage; IL: Interleukin; TNF: Tumor necrosis factor.