Electroacupuncture Enhance Therapeutic Efficacy of Mesenchymal Stem Cells Transplantation in Rats With Intracerebral Hemorrhage

Abstract

Background Previous studies have showed the bene cial effects of mesenchymal stem cells (MSCs) on experimental intracerebral hemorrhage (ICH) animal. Enhancement of the treatment e cacy of MSCs in ICH is essential, considering the diseases association with high rates of disability and mortality. Some auxiliary methods to enhance the bene cial e cacy of MSCs have been introduced. However, the effect of electroacupuncture (EA) on the therapeutic e cacy of MSCs transplantation in hemorrhagic stroke and its potential mechanism is not explored. Methods ICH rat models were established using collagenase and heparin. 48 h after ICH induction, the rats were randomly divided into model control (MC), MSCs transplantation (MSCs), EA stimulation (EA) and MSCs transplantation combined with EA stimulation (MSCs + EA) groups. We used mNSS test and gait analysis to assess neurological function of rats, and PET/CT to evaluate the volume of hemorrhage focus and level of glucose uptake. The concentrations of MDA, SOD, NSE, S100B and MBP in serum or plasma were examined with ELISA. Neural differentiation of MSCs, and the expressions of Bcl-2, Bax, Arg-1 and iNOS proteins around hematoma were detected by immuno uorescence and immunohistochemistry staining respectively. Western blot was carried out to analyze the expression levels of COX4, OGDH, PDH-E1α, Bcl-2 and Bax proteins. TUNEL staining was used to estimate cell apoptosis and transmission electron microscopy (TEM) was used to observe the ultrastructure and number of mitochondria. Results Our data showed that EA promoted neuron-like differentiation of transplanted MSCs and the expressions of BDNF and NGF proteins in ICH rats. The score of mNSS and the gait analysis showed that the recovery of the neurological function in the MSCs + EA group was better than that in the MSCs and EA groups. EA improved the structures of brain tissue, and alleviated brain injury further after MSCs transplantation in ICH rats. When compared with the MSCs and EA groups, the level of glucose uptake and numbers of mitochondria and Arg-1 positive cells in MSCs + EA group increased signi cantly, but the numbers of apoptotic cells and iNOS positive cells and volume of hemorrhage focus reduced. The expressional levels of COX4, OGDH, PDH-E1α and Bcl-2 proteins increased, while the expressional levels of Bax protein decreased compared with those in the MSCs and EA groups. Conclusion Our results reveal that EA improve therapeutic e cacy of MSCs transplantation in ICH rats.

Mesenchymal Stem Cell-Derived Neuron-Like Cell Transplantation Combined with Electroacupuncture Improves Synaptic Plasticity in Rats with Intracerebral Hemorrhage via mTOR/p70S6K Signaling

“…Previous studies have indicated that the transplantation of MSCs combined with EA stimulation promotes axonal regeneration and functional recovery of the injured spinal cord [ 24 , 25 ] and leads to a better therapeutic effect by increasing the expression of neurotrophic factors, regulating neurogenesis, and increasing the neural differentiation of transplanted cells in ischemic stroke compared with a single therapy [ 26 , 27 ]. We also confirmed that the combined treatment improves neurological function in rats with ICH in a previous study [ 28 ]. However, the exact mechanism by which MSCs combined with EA improve neurological function remains to be further explored.…”

“…Rat MSCs labeled with green fluorescent protein (GFP) (Cyagen Biosciences, China) [ 28 ] were removed from liquid nitrogen, thawed in a 37°C water bath, and then centrifuged. After centrifugation, MSCs were resuspended in alpha-minimum essential medium ( α -MEM) (HyClone) supplemented with 10% fetal bovine serum (FBS, HyClone), 100 mg/ml streptomycin, and 100 U/ml penicillin and incubated in a humidified atmosphere with 5% CO 2 at 37°C.…”

Mesenchymal Stem Cell Application and Its Therapeutic Mechanisms in Intracerebral Hemorrhage

“…Moreover, EA downregulated the expression of neurogenesis inhibitory molecules, including NogoA, lysophosphatidic acid, and RhoA, to improve the proliferation and differentiation of endogenous neural stem cells and synaptic plasticity in stroke rats ( Tan et al, 2018 ; Yang et al, 2022 ). Deng et al (2021) have reported that EA promoted differentiation of transplanted MSCs into neuron-like cells and expressions of BDNF and NGF proteins displaying therapeutic efficacy in ICH rats. Furthermore, Zhang et al (2015) suggested that the combination of direct stereotactic intracerebral injection of HUC-MSCs and minimally invasive hematoma aspiration was better than either therapy alone in reducing neuronal damage and improving neuronal functions.…”

Electroacupuncture reduces oxidative stress response and improves secondary injury of intracerebral hemorrhage in rats by activating the peroxisome proliferator-activated receptor-γ/nuclear factor erythroid2-related factor 2/γ-glutamylcysteine synthetase pathway

“…With the common progress of acupuncture and evidence-based medicine, the clinical and basic researches on the treatment of ICH by EA continue to emerge, and the interpretation of its mechanism is also constantly improving. EA can exert neuroprotective effect on brain injury after ICH through many ways, such as anti-inflammation, antioxidation, antiapoptosis and increasing brain derived neurotrophic factor protein [17–19]. In this study, we found that EA could activate PPARγ/Nrf2/γ-GCS signaling pathway, inhibit microglia activation, reduce ROS production, and accelerate ROS clearance, thus alleviating oxidative stress response and the extent of myelin sheath damage, improve neurological function after ICH.…”