Scalp Acupuncture and Treadmill Training Inhibits Neuronal Apoptosis through Activating cIAP1 in Cerebral Ischemia Rats

Abstract

Stroke is the leading cause of long-term disability in developed countries. Multitudinous evidence suggests that treadmill training treatment is beneficial for balance and stroke rehabilitation; however, the need for stroke therapy remains unmet. In the present study, a cerebral ischemia rat model was established by permanent middle cerebral artery occlusion (pMCAO) to explore the therapeutic effect and mechanism of scalp acupuncture combined with treadmill training on ischemic stroke. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and neuronal nuclear protein (NeuN) double staining and cellular inhibitor of apoptosis protein-1 (cIAP1) and NeuN immunofluorescence double staining were used to detect the short-term and long-term neuroprotective effects of scalp acupuncture combined with treadmill training on pMCAO rats. In addition, the antiapoptotic effect of the combined treatment was evaluated in pMCAO rats transfected with cIAP1 shRNA. Western blotting was used to detect the relative protein expression in the caspase-8/-9/-3 activation pathway downstream of cIAP1 to further clarify its regulatory mechanism. Our results showed that scalp acupuncture combined with treadmill training successfully achieved short-term and long-term functional improvement within 14 days after stroke, significantly inhibited neuronal apoptosis, and upregulated the expression of cIAP1 protein in the ischemic penumbra area of the ischemic brain. However, no significant functional improvement and antiapoptotic effect were found in pMCAO rats transfected with cIAP1 shRNA. Western blotting results showed that the combined therapy markedly inhibited the activation of the caspase-8/-9/-3 pathway. These findings indicate that scalp acupuncture combined with treadmill training therapy may serve as a more effective alternative modality in the treatment of ischemic stroke, playing an antiapoptotic role by upregulating the expression of cIAP1 and inhibiting the activation of the caspase-8/-9/-3 pathway.

Acupuncture for stroke: A bibliometric analysis of global research from 2000 to 2022

“…Acupuncture can inhibit excessive apoptosis of neurons to maintain the number of cells, generate new neurons to replace apoptotic neurons, and accelerate the process of neurogenesis [ 62 ]. Acupuncture promotes the expression of many growth factors, such as glial cell line-derived neurotrophic factor (GDNF), stromal cell-derived factor-1α (SDF-1α) [ 63 ], brain-derived neurotrophic factor (BDNF), and basic fibroblast growth factor (bFGF) [ 64 ], enabling neuronal survival and angiogenesis after stroke [ 65 ]. It also promotes neurogenesis and inhibits apoptosis.…”

Effects of acupuncture on earthquake survivors with major psychiatric disorders and related symptoms: A scoping review of clinical studies

“…Thus far, scalp electro-acupuncture has been reported to have beneficial effects on various neuropsychiatric diseases, including vascular dementia [ 45 ], autism [ 46 ], stroke [ 47 ], post-stroke depression [ 48 ], and MDD [ 49 ]. Although the underlying therapeutic mechanism by which scalp electro-acupuncture promotes beneficial effects against various neuropsychiatric disorders needs to be further elucidated, this approach has the potential for neuroprotective effects [ 50 , 51 ] and modulating the functional connectivity between brain regions [ 52 ]. According to our findings, EX-HN1 and GV24 were the main acupoints deserving attention in this field.…”

Influence of metabolic stress and metformin on synaptic protein profile in SH‐SY5Y‐derived neurons

“…Total raptor (Cell Signaling #2280 (Sanders et al., 2022)) and phosphorylated raptor (S792, Cell Signaling #2083 (Sanders et al., 2022)) levels were also measured alongside total P70S6K (Cell Signaling #9202 (Zhang et al., 2021)), phosphorylated P70S6K (Santa Cruz SC‐11759 (Xiao et al., 2002)), total ULK (Cell signaling #8054 (Wu et al., 2020)), and ULK S555 (cell signaling #5869 (Wu et al., 2020)). Synaptophysin (Cell Signaling #5461 (Wu et al., 2018)), NeuN (Cell Signaling #24307 (Tang et al., 2021)), PSD‐95 (post‐synaptic marker, Cell Signaling #36233 (Shui et al., 2022)), Homer‐1 (post‐synaptic marker, SC‐136358 (Wang et al., 2014)), SNAP‐25 (Cell signaling #5308 (Polishchuk et al., 2023)), VAMP2 (Cell signaling #13508 (Arrojo et al., 2019)) were measured as synaptic and neuronal markers. Total IR α (Cell signaling #74118 (Dall'Agnese et al., 2022)), total IR β (Cell signaling #23413 (Dall'Agnese et al., 2022)), IRS1 S636 (Cell signaling #2388 (Uddin et al., 2019)), total IRS1 (Cell signaling #2390 (Tang et al., 2019)), mTOR S2448 (Cell signaling #2971 (Luo et al., 2022)), total mTOR (Cell signaling #2972 (Luo et al., 2022)), Akt T308 (Cell signaling #9275 (Mathieu et al., 2019)), Akt S473 (Cell signaling #4058 (Marko et al., 2020)), and total Akt (Cell signaling #4685 (Marko et al., 2020)) were measured as markers of insulin signaling.…”

“…Total raptor (Cell Signaling #2280 (Sanders et al, 2022)) and phosphorylated raptor (S792, Cell Signaling #2083 (Sanders et al, 2022)) levels were also measured alongside total P70S6K (Cell Signaling #9202 (Zhang et al, 2021)), phosphorylated P70S6K (Santa Cruz SC-11759 (Xiao et al, 2002)), total ULK (Cell signaling #8054 (Wu et al, 2020)), and ULK S555 (cell signaling #5869 (Wu et al, 2020)). Synaptophysin (Cell Signaling #5461 (Wu et al, 2018)), NeuN (Cell Signaling #24307 (Tang et al, 2021)), PSD-95 (post-synaptic marker, Cell Signaling #36233 (Shui et al, 2022)), Homer-1 (post-synaptic marker, SC-136358 (Wang et al, 2014) (Tang et al, 2019)), mTOR S2448 (Cell signaling #2971 (Luo et al, 2022)), total mTOR (Cell signaling #2972 (Luo et al, 2022)), Akt T308 (Cell signaling #9275 (Mathieu et al, 2019)), Akt S473 (Cell signaling #4058 (Marko et al, 2020)), and total Akt (Cell signaling #4685 (Marko et al, 2020)) were measured as markers of insulin signaling. Ponceau S, α-tubulin (Cell signaling #2144 (Mertins et al, 2021)), GAPDH (Abcam ab8245 (Luo et al, 2023)) or β-Actin (Abcam ab8227 (Moll et al, 2023)) were utilized as loading controls.…”