Bone marrow mesenchymal stem cells upregulate PI3K/AKT pathway and down-regulate NF-κB pathway by secreting glial cell-derived neurotrophic factors to regulate microglial polarization and alleviate deafferentation pain in rats.

Deafferentation pain (DP), a typical neuropathic pain, occurs due to peripheral or central sensory nerve injury, which causes abnormal discharge of the upstream neurons or C fibers. Current treatment methods for DP have multiple side effects. Bone marrow mesenchymal stem cells (BMSC) have been used to treat neuropathic pain because of their ability to regulate neuroinflammation. Glial cell-derived neurotrophic factor (GDNF) is a neurotrophic mediator that exerts neuroprotective effects in neurological diseases. In this study, we investigated whether DP could be alleviated by BMSCs and the underlying mechanism. In vitro study, microglia was stimulated by lipopolysaccharide and then co-cultured with BMSC, GDNF or siRNA GDNF-BMSC. In vivo study, BMSC or siRNA GDNF-BMSC was transplanted intramedullarily on the 21st day after DP surgery. The expression of inflammatory-related factors were detected by RT-PCR and ELISA, RT-PCR,flow cytometry and immunofluorescence staining were performed to detect the expression of microglial surface markers, and Western blot was used to detect the expression levels of p-NF-kb, pPI3K, and pAKT. The pain-related behavioral changes were detected 7 days after transplantation. ELISA and RT-PCR results showed that the production of inflammatory cytokines in lipopolysaccharide-stimulated microglia and DP model plasma was downregulated, while anti-inflammatory mediators were upregulated significantly following pretreatment with BMSCs or GDNF. Flow cytometry, immunofluorescence staining, and RT-PCR results showed that BMSCs inhibited the microglial M1 phenotype and promoted the M2 phenotype by secreting GDNF. Furthermore, modulation functions of BMSCs involve inhibiting NF-κB while promoting PI3K /AKT signaling pathway activation. We found that our in vivo DP model was completely deafferent and BMSC administration clearly alleviated symptoms of DP. This function was also, at least partly, achieved by GDNF. The present studies demonstrate that BMSC can inhibit neuroinflammation by transforming microglial destructive M1 phenotype into regenerative M2 phenotype, and thus alleviate DP,likely by suppressing the NF-κB signaling pathway while promoting the PI3K/AKT signaling pathway activation through producing GDNF. The present findings are in support of the potential therapeutic application of BMSCs and the pharmaceutical application of GDNF for DP.

Zhong Z ，Chen A ，Fa Z ，Ding Z ，Xiao L ，Wu G ，Wang Q ，Zhang R ... -  《-》

Anti-inflammatory protein TSG-6 secreted by bone marrow mesenchymal stem cells attenuates neuropathic pain by inhibiting the TLR2/MyD88/NF-κB signaling pathway in spinal microglia.

Yang H ，Wu L ，Deng H ，Chen Y ，Zhou H ，Liu M ，Wang S ，Zheng L ，Zhu L ，Lv X ... -  《Journal of Neuroinflammation》

Bone marrow-mesenchymal stem cells alleviate microglial Pyroptosis after intracerebral hemorrhage in rat by secreting C1q/tumor necrosis factor-related protein 3.

There are no specific treatment methods for intracerebral hemorrhage (ICH). Neuroinflammation triggered by microglial pyroptosis plays an important role in ICH pathophysiology. Bone marrow mesenchymal stem cells (BMSCs) are widely used in the treatment of neurological diseases because of their paracrine function. In this study, we aimed to clarify whether BMSCs can alleviate microglial pyroptosis after ICH by secreting C1q/tumor necrosis factor-related protein 3 (CTRP3), a adiponectin paralog with established metabolic regulatory properties and neuroprotective effects. In an in vitro study, microglia were stimulated with hemin for pyroptosis and then co-cultured with BMSCs, CTRP3, or CTRP3-small interfering RNA (siRNA)-BMSC; in an in vivo study, intracerebroventricular transplantation of BMSCs or siRNA-CTRP3-BMSCs was performed after ICH surgery. The expression of inflammation-related factors was detected by qRT-PCR and ELISA. Western blotting and immunofluorescence staining were performed to detect the expression of pyroptotic protein, and western blotting was used to detect the activation of phosphoinositide 3-kinase (PI3K), protein kinase B (AKT) and splenic tyrosine kinase (Syk). Behavioral changes were detected 7 days after transplantation. ELISA and qRT-PCR results showed that the production of inflammatory cytokines in hemin-stimulated microglia was significantly downregulated following pretreatment with BMSCs or CTRP3. The Caspase-1 activity assay kit and western blotting results showed that BMSCs attenuated microglial pyroptosis by secreting CTRP3. Furthermore, the modulation functions of BMSCs or CTRP3 involve the promotion of PI3K/AKT and inhibition of Syk signaling pathway activation. Neurological deficits, edema, and disruption of tight junction protein were completely alleviated, while inflammation-related factors and microglial pyroptosis after ICH were significantly downregulated after BMSCs administration. BMSCs can inhibit neuroinflammation by inhibiting microglial pyroptosis, thus alleviate ICH symptoms, likely by suppressing the Syk signaling pathway while promoting the PI3K/AKT signaling pathway activation through producing CTRP3.

Wang Q ，Piao J ，Li Y ，Tu H ，Lv D ，Hu L ，Zhang R ，Zhong Z ... -  《-》

Adipose-Derived Stem Cells Modulate BV2 Microglial M1/M2 Polarization by Producing GDNF.

Zhong Z ，Chen A ，Fa Z ，Ding Z ，Xie J ，Sun Y ，Zhang R ，Wang Q ... -  《-》

Exosomal GDNF from Bone Marrow Mesenchymal Stem Cells Moderates Neuropathic Pain in a Rat Model of Chronic Constriction Injury.

Both of exosomes derived from mesenchymal stem cells (MSCs) and glial cell line-derived neurotrophic factor (GDNF) show potential for the treatment of neuropathic pain. Here, the analgesic effects of exosomes derived from bone marrow MSCs (BMSCs) were investigated. BMSCs-derived exosomes were isolated and characterized. Chronic constriction injury (CCI) was constructed to induce neuropathic pain in rats, which were then treated with exosomes. Pain behaviors were evaluated by measuring paw withdrawal thresholds and latency. The changes of key proteins, including cytokines, were explored using Western blot and ELISA. Administration of BMSCs-derived exosomes alleviated neuropathic pain, as demonstrated by the decrease of thermal hyperalgesia and mechanical allodynia, as well as the reduced secretion of pro-inflammatory cytokines in CCI rats. These effects were comparable to the treatment of GDNF alone. Mechanically, the exosomes suppressed the CCI-induced activation of TLR2/MyD88/NF-κB signaling pathway, while GDNF knockdown impaired their analgesic effects on CCI rat. BMSCs-derived exosomes may alleviate CCI-induced neuropathic pain and inflammation in rats by transporting GDNF.

Zhang X ，Liu H ，Xiu X ，Cheng J ，Li T ，Wang P ，Men L ，Qiu J ，Jin Y ，Zhao J ... -  《-》