Ganglioside Treatment Research Articles

Diammonium glycyrrhizinate promotes the regeneration and repair of central nervous system in rats with severe traumatic brain injury by Wnt/β-catenin signaling pathway

To observe the effects of diammonium glycyrrhizinate (DG) on nerve regeneration repair in rats with severe traumatic brain injury (STBI) from the perspective of Wnt/β-catenin signaling pathway. Seventy-two Sprague-Dawle (SD) male rats were randomly divided into normal group, STBI model group, ganglioside (GA) treatment group and DG treatment group. The STBI animal model was reproduced referring to modified Feeney free fall impact model. No injury was made in normal group. Six hours after modeling, monosialotetrahexosylganglioside sodium injection and DG injection were injected via tail vein of rats in GA treatment group and DG treatment group respectively, once a day for 7 days. Normal group and STBI model group were given the same amount of normal saline. Six rats in each group were sacrificed on the 1st, 3rd and 7th day after the challenge for neurological severity score (NSS), and then the blood of abdominal aorta was drawn and brain tissue was harvested. The contents of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in serum were detected by enzyme linked immunosorbent assay (ELISA). The pathological changes of sub-granular zone (SGZ) were observed under light microscope after hematoxylin eosin (HE) staining. Real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to detect the mRNA expressions of Wnt3a, β-catenin, glycogen synthetase kinase-3β (GSK-3β) and Axin. (1) There was no neurological deficit in the normal group and NSS was 0. NSS score of rats increased significantly on the first day after modeling, and then decreased gradually over time. NSS of the rats treated with GA and DG were significantly lower than that of the STBI model rats (score: 7.33±2.07, 6.17±2.23 vs. 9.33±1.63, both P < 0.01). Though NSS gradually decreased over time, the differences were still statistically significant on the 7th day (score: 2.67±0.82, 1.00±0.00 vs. 6.17±2.23, both P < 0.01), and NSS of DG treatment group was significantly lower than that of GA treatment group. (2) In SGZ of rats, cells were arranged in a compact and orderly way in the normal group, but neurons and tissues were damaged and destroyed at different time points in the STBI model group. After either GA or DG treatment, the damage of nerve tissue was improved gradually over time, and the effect of DG was more obvious. (3) In the normal group, the mRNA expressions of Wnt3a and β-catenin were almost not expressed, the mRNA expressions of GSK-3β and Axin were higher, and the contents of BDNF and NGF in serum were less. On the 1st day after STBI, the mRNA expressions of Wnt3a and β-catenin in hippocampus, the contents of BDNF and NGF in serum were significantly increased, and the mRNA expressions of GSK-3β and Axin were significantly decreased. The mRNA expressions of Wnt3a and β-catenin in the hippocampus and the contents of BDNF and NGF in serum were significantly higher than those in the model group 1 day after GA or DG was added, the mRNA expressions of GSK-3β and Axin were significantly decreased, and the effect of DG was more significant than that of GA [Wnt3a mRNA (2-ΔΔCt): 3.51±0.14 vs. 2.93±0.05, β-catenin mRNA (2-ΔΔCt): 1.90±0.08 vs. 1.75±0.04, BDNF (ng/L): 4.06±0.55 vs. 3.16±0.64, NGF (ng/L): 9.53±1.08 vs. 7.26±0.43, GSK-3β mRNA (2-ΔΔCt): 0.75±0.01 vs. 0.79±0.01, Axin mRNA (2-ΔΔCt): 0.74±0.02 vs. 0.76±0.02, all P < 0.05]. It was gradually increasing or decreasing over time and the difference was still statistically significant up to the 7th day. DG can promote the recovery of nerve function in rats with STBI, and its mechanism may be related to the regeneration of nerve cells proliferation and differentiation by Wnt/β-catenin signaling pathway and the reconstruction of nerve tissue in SGZ of hippocampus.

Clinical efficacy of gangliosides on premature infants suffering from white matter damage and its effect on the levels of IL-6, NSE and S100β.

This study investigated the clinical efficacy of gangliosides on premature infants suffering from white matter damage and its effect on the levels of IL-6, neuron-specific enolase (NSE) and S100β. Seventy-six cases of premature infants suffering from white matter damage admitted to the Tianjin Central Hospital of Obstetrics and Gynecology from February 2016 to March 2017 were enrolled in this study. They were randomly divided into the control group and the observation group with 38 cases in each group. Control group was given conventional treatment, while the observation group was given ganglioside treatment on the basis of the treatment given to the control group. Craniocerebrum ultrasonic detection was used to observe the condition of white matter around the ventricle of child patients in the two groups, before and after treatment. ELISA was used to detect the levels of IL-6, NSE and S100β. Gesell developmental scale was used to compare the developmental quotient (DQ) of various function regions of the children. The total effective rate of the observation group was higher than that of the control group (P<0.05). The gray value of craniocerebrum ultrasonic detection in the observation group was significantly lower than that in the control group (P<0.05). IL-6, S100β and NSE levels of the child patients in the two groups were significantly declined at 7 and 14 days after birth (P<0.05). After 1 year, the observation group scored significantly higher DQ than the control group in the aspects of social adaptation, gross motor, fine motor, language and personal social contact. The sequel incidence of patients in the observation group was significantly lower than that of the control group (P<0.05). In conclusion, the intervention treatment with ganglioside for premature infants suffering from white matter damage was beneficial and provided a protective effect. It also reduced sequel and produced some promising results.

Clinical Characteristics of Intravenous Injection of Monosialotetrahexosyl Ganglioside Sodium-Related Guillain-Barre Syndrome.

Introduction: Guillain Barre Syndrome (GBS) is an acute inflammatory immune-mediated multiple nerve root neuropathy. GBS primarily damages the spinal nerve root and peripheral nerves, but can also affect the cranial nerves and cause acute demyelination. This study analyzed the clinical features of intravenous injection of monosialotetrahexosyl ganglioside sodium-related Guillain-Barre syndrome (GRD-GBS).Methods: We retrospectively studied 12 patients who developed GRD-GBS after receiving monosialotetrahexosyl ganglioside sodium treatment in association with recent trauma, surgery, acute cerebrovascular disease, or chronic peripheral neuropathy. Clinical characteristics, electrophysiological examinations, serum-specific antibodies, and prognosis were assessed. As controls, we selected 12 patients hospitalized with non-ganglioside-related (NGRD)-GBS.Results: The positive rate of the ganglioside antibody test was significantly higher in the GRD-GBS group (66.67%) than in the NGRD-GBS group (8.33%). CSF protein levels were similar between the groups, but the incidence of blood-nerve-barrier (BNB) disruption was higher in the GRD-GBS group. Patient scores for the Hughes Functional Grading Scale (HFGS), a disability scale, were higher (more severe disability) in the GRD-GBS group than in the NGRD-GBS group. The HFGS scores of the GRD-GBS group did not change between peak onset and 30 days after discharge, but did change significantly by 90 days after discharge, while scores were significantly lower at both 30 and 90 days after discharge in the NGRD-GBS group.Conclusions: GRD-GBS patients showed more severe clinical manifestations, poorer prognosis, and slower recovery than patients with NGRD-GBS. Ganglioside treatment should be used with extreme caution in patients with trauma that damages the BNB.

Meta-analysis evaluation of the treatment of neonatal hypoxic-ischemic encephalopathy with ganglioside

To evaluate the efficacy and safety of ganglioside in the treatment of neonates who suffer from hypoxic-ischemic encephalopathy(HIE). To research the databases of PubMed, ScienceDirect, LISTA, CNKI, Chinese biomedical literature database, Wanfang digital journals of full-text database, then to determine the inclusion and exclusion criteria and conduct the Meta-analysis with RevMan5.0 software after quality evaluation of literatures. A total of 12 papers were included, the results of Meta-analysis showed that compared with the control group, the abnormal rate declined in the ganglioside-treat group(relative risk(RR)=0.27, 95% confidence interval (CI)= 0.05-1.96)NBNA records of the 7d,10-14d improved effectively with the RR (95%CI) were 2.28(0.86-3.42), 2.53(1.04-2.92)respectively. neural system sequelae incidence is reduced significantly(RR=0.35, 95%CI: 0.15-0.79). Ganglioside treatment could reduce the head CT abnormality rate effectively , improve the neurological score, reduce the incidence of neurological sequelae, and prompt clinical recovery significantly for neonates with HIE when compared with the control group.

An in vivo study of hypoxia-inducible factor-1α signaling in ginsenoside Rg1-mediated brain repair after hypoxia/ischemia brain injury

Hypoxia/ischemia (HI) brain injury is a common central nervous system insult in newborns. Studies have demonstrated bioactivity of ginsenoside Rg1 in increasing neural viability and promoting angiogenesis. However, there are few reports on roles of Rg1 in brain repair of neonatal HI, and the mechanisms involved are unclear. a neonatal HI model was established by a modified Rice-Vannucci model (RVM) and pups received ginsenoside Rg1 or monosialotetrahexosyl ganglioside (GM1) treatment. Neurological function and pathologic damage of rats were evaluated. Cellular apoptosis was detected with Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Immunohistochemistry for von willebrand factor (vwf) was used to label micro vessels. Expression levels of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), and cleaved caspase 3 (CC3) were detected by western blot. Both Rg1 and GM1 reduced neurological impairment and pathologic damage after HI by enhancing neural survival. Rg1, but not GM1, could also facilitate angiogenesis after HI. These pharmacological effects of Rg1 may be attributed to regulation of expression level of VEGF and CC3 and HIF-1α signaling pathway was involved. Rg1 plays a neuroprotective role in brain repair following neonatal HI, and HIF-1α is a potential target for therapeutic intervention in neonates with HI brain injury.

HAM/TSP patient treated with porcine cortex monosialoganglioside: clinical outcome and follow up

HTLV-1 infects about 20 million people worldwide and is considered the etiologic agent of Adult T-cell Leukemia/Lymphoma (ATLL) and HTLV-1 Associated Mielopathy/Tropical Spastic Paraparesis (HAM/TSP). There is still no effective therapy for none of the pathologies. Porcine cerebral cortex GM1 monosialoganglioside (SINAXIAL®) is being prescribed for traumatic or acute peripheral neuropathies, acute cerebrovascular and others. CD4 modulation and inhibition of HIV-1 infectivity induced by monosialoganglioside GM1 in vitro has been published although the effect was not long-lasting since removal of the compound was followed by a rapid increase in viral replication. The aim of this report is to describe the case of an HTLV-1 infected woman with HAM/TSP who was treated with GM1 for 6 months. The 40 year old individual was from Peru and started showing HAM/TSP symptoms (leg weakness, numbness, pain at the lower limbs and back) in 2010. In April 2013, she received Pregabalin (75 mg), although no improvement was observed. In December, the patient started to receive GM1, injected locally in her legs, according to the following schedule: 7 monthly applications combined with Acetilcarnitine and Citicoline (5 mg) followed by 3 months of resting. After 6 months, the treatment was suspended. Three months later, the patient kept on going with the Acetilcarnitine and Citicoline treatment. However, symptoms started to increase. The patient's description of the symptoms’ severity were consistent with proviral load (PVL) levels, quantified two months before the GM1 ganglioside treatment and 1, 5 and 8 months after. It could be suggested that porcine cerebral cortex GM1 monosialoganglioside applied monthly had positive short term effects reflected both in the amelioration of the symptoms and the PVL decrease. Further studies should be carried out in order to establish the role of ganglioside in HAM/TSP treatment.

GBM Derived Gangliosides Induce T Cell Apoptosis through Activation of the Caspase Cascade Involving Both the Extrinsic and the Intrinsic Pathway.

Previously we demonstrated that human glioblastoma cell lines induce apoptosis in peripheral blood T cells through partial involvement of secreted gangliosides. Here we show that GBM-derived gangliosides induce apoptosis through involvement of the TNF receptor and activation of the caspase cascade. Culturing T lymphocytes with GBM cell line derived gangliosides (10-20μg/ml) demonstrated increased ROS production as early as 18 hrs as indicated by increased uptake of the dye H2DCFDA while western blotting demonstrated mitochondrial damage as evident by cleavage of Bid to t-Bid and by the release of cytochrome-c into the cytosol. Within 48-72 hrs apoptosis was evident by nuclear blebbing, trypan blue positivity and annexinV/7AAD staining. GBM-ganglioside induced activation of the effector caspase-3 along with both initiator caspases (-9 and -8) in T cells while both the caspase-8 and -9 inhibitors were equally effective in blocking apoptosis (60% protection) confirming the role of caspases in the apoptotic process. Ganglioside-induced T cell apoptosis did not involve production of TNF-α since anti-human TNFα antibody was unable to protect T cells from nuclear blebbing and subsequent cell death. However, confocal microscopy demonstrated co-localization of GM2 ganglioside with the TNF receptor and co-immunoprecipitation experiments showed recruitment of death domains FADD and TRADD with the TNF receptor post ganglioside treatment, suggesting direct interaction of gangliosides with the TNF receptor. Further confirmation of the interaction between GM2 and TNFR1 was obtained from confocal microscopy data with wild type and TNFR1 KO (TALEN mediated) Jurkat cells, which clearly demonstrated co-localization of GM2 and TNFR1 in the wild type cells but not in the TNFR1 KO clones. Thus, GBM-ganglioside can mediate T cell apoptosis by interacting with the TNF receptor followed by activation of both the extrinsic and the intrinsic pathway of caspases.

More severe manifestations and poorer short-term prognosis of ganglioside-associated Guillain-Barré syndrome in Northeast China.

Ganglioside as a neurotrophic drug has been hitherto widely used in China, although Guillain-Barré syndrome (GBS) following intravenous ganglioside treatment was reported in Europe several decades ago. We identified 7 patients who developed GBS after intravenous use of gangliosides (ganglioside+ group) and compared their clinical data with those of 77 non-ganglioside-associated GBS patients (ganglioside− group) in 2013, aiming at gaining the distinct features of ganglioside-associated GBS. Although the mean age, protein levels in cerebrospinal fluid (CSF) and frequency of cranial nerve involvement were similar between the two groups, the Hughes Functional Grading Scale (HFGS) score and the Medical Research Council (MRC) sum score at nadir significantly differed (4.9±0.4 vs 3.6±1.0; 7.7±5.5 vs 36.9±14.5, both p<0.001), indicating a higher disease severity of ganglioside-associated GBS. A higher ratio of patients with ganglioside-associated GBS required mechanical ventilation (85.7% vs 15.6%, p<0.01). The short-term prognosis of ganglioside-associated GBS, as measured by the HFGS score and the MRC sum score at discharge, was poorer (4.3±0.5 vs 2.8±1.1; 17.3±12.9 vs 46.0±13.9, both p<0.001). All the patients in the ganglioside+ group presented an axonal form of GBS, namely acute motor axonal neuropathy (AMAN). When compared with the AMAN patients in the ganglioside− group, more severe functional deficits at nadir and poorer recovery after standard treatment were still prominent in ganglioside-associated GBS. Anti-GM1 and anti-GT1a antibodies were detectable in patients with AMAN while not in patients with the demyelinating subtype of GBS. The concentrations of these antibodies in patients with AMAN were insignificantly different between the ganglioside+ and ganglioside− groups. In sum, ganglioside-associated GBS may be a devastating side effect of intravenous use of gangliosides, which usually manifests a more severe clinical course and poorer outcome.

Lower motor neuron syndrome associated with IgG anti-GM1 antibodies revisited

A patient, who developed an amyotrophic lateral sclerosis-like disorder subsequent to ganglioside treatment, had IgM antibodies to GM2 as well as to minor gangliosides X1 and X2 containing GM2 epitope. These gangliosides as well as GM1 were tested in 655 sera obtained from patients who were suspected of having amyotrophic lateral sclerosis or motor neuron disease to find a treatable condition. Three patients had high titers of IgG anti-GM1 antibodies, but no IgM anti-GM1 antibodies. One of the patients also had IgG anti-X2 antibodies. The patients, being diagnosed with having lower motor neuron syndrome, had neither upper motor neuron signs nor multifocal conduction block. Both IgM and IgG anti-GM1 antibodies should be tested in patients who have lower motor neuron syndrome.

Therapeutic effect of human umbilical cord mesenchymal stem cells on neonatal rat hypoxic–ischemic encephalopathy

The therapeutic potential of umbilical cord blood mesenchymal stem cells has been studied in several diseases. However, the possibility that human umbilical cord Wharton's jelly-derived mesenchymal stem cells (hUCMSCs) can be used to treat neonatal hypoxic-ischemic encephalopathy (HIE) has not yet been investigated. This study focuses on the potential therapeutic effect of hUCMSC transplantation in a rat model of HIE. Dermal fibroblasts served as cell controls. HIE was induced in neonatal rats aged 7 days. hUCMSCs labeled with Dil were then transplanted into the models 24 hr or 72 hr post-HIE through the peritoneal cavity or the jugular vein. Behavioral testing revealed that hUCMSC transplantation but not the dermal fibroblast improved significantly the locomotor function vs. vehicle controls. Animals receiving cell grafts 24 hr after surgery showed a more significant improvement than at 72 hr. More hUCMSCs homed to the ischemic frontal cortex following intravenous administration than after intraperitoneal injection. Differentiation of engrafted cells into neurons was observed in and around the infarct region. Gliosis in ischemic regions was significantly reduced after hUCMSC transplantation. Administration of ganglioside (GM1) enhanced the behavioral recovery on the base of hUCMSC treatment. These results demonstrate that intravenous transplantation of hUCMSCs at an early stage after HIE can improve the behavior of hypoxic-ischemic rats and decrease gliosis. Ganglioside treatment further enhanced the recovery of neurological function following hUCMSC transplantation.

The ganglioside treatment and rehabilitation care in early stage after traumatic brain injury

Objective To observe ganglioside and rehabilitation care to early rehabilitation of patients with severe traumatic brain injury.Methods 66 patients with brain injury were randomly divided into the control group and the rehabilitation group with 33 cases in each group.Both groups were given conventional medication and ganglioside treatment.The control group was given routine care.In addition to the usual care,the rehabilitation group received the implementation of systematic and standardized early rehabilitation nursing.Oxidative stress status,Barthel index score,and quality of life were compared between two groups.Results Oxidative stress parameters of the two groups showed no significant difference.After hospital treatment,Barthel index score and quality of life in the rehabilitation group were significantly higher than those of the control group.Conclusions Rehabilitation care can reduce disability rates of head injury,and improve the quality of life of patients. Key words: Traumatic brain injury; Gangliosides; Oxidative stress status; Rehabilitation care; Quality of life; Barthel index

Chagas' cardioneuropathy: Effects of ganglioside treatment on stress-induced arrhythmias. A randomized, double-blind, placebo-controlled study

Chagas’ disease is manifested by cardiovascular and autonomic nervous system disorders with arrhythmias and sudden death.

Ganglioside-exposed dendritic cells inhibit T-cell effector function by promoting regulatory cell activity

Tumour pathogenesis is characterized by an immunosuppressive microenvironment that limits the development of effective tumour-specific immune responses. This is in part the result of tumour-dependent recruitment and activation of regulatory cells, such as myeloid-derived suppressor cells and regulatory T cells in the tumour microenvironment and draining lymph nodes. Shedding of gangliosides by tumour cells has immunomodulatory properties, suggesting that gangliosides may be a critical factor in initiating an immunosuppressive microenvironment. To better define the immunomodulatory properties of gangliosides on antigen-specific T-cell activation and development we have developed an in vitro system using ganglioside-treated murine bone-marrow-derived dendritic cells to prime and activate antigen-specific CD4(+) T cells from AND T-cell receptor transgenic mice. Using this system, ganglioside treatment promotes the development of a dendritic cell population characterized by decreased CD86 (B7-2) expression, and decreased interleukin-12 and interleukin-6 production. When these cells are used as antigen-presenting cells, CD4 T cells are primed to proliferate normally, but have a defect in T helper (Th) effector cell development. This defect in Th effector cell responses is associated with the development of regulatory T-cell activity that can suppress the activation of previously primed Th effector cells in a contact-dependent manner. In total, these data suggest that ganglioside-exposed dendritic cells promote regulatory T-cell activity that may have long-lasting effects on the development of tumour-specific immune responses.

Tumor derived gangliosides promote regulatory cell activity (98.10)

Abstract Gangliosides, shed by tumor cells, have been shown to have immunomodulatory properties, suggesting that gangliosides may be a critical factor in initiating an immunosuppressive tumor microenvironment. To better define the immunomodulatory properties of gangliosides on antigen-specific T cell activation and development, independent of other tumor associated factors, we have developed an in vitro system utilizing ganglioside-treated murine bone marrow derived dendritic cells to prime and activate antigen-specific CD4+ T cells from AND T cell receptor transgenic mice. Ganglioside treatment promotes the development of a regulatory DC population characterized by decreased CD86 expression, and decreased IL-12 and IL-6 production. CD4 T cells primed with ganglioside-treated DCs proliferate normally, but have a defect in Th1 effector cell development and IL-2 production. This defect in Th1 effector cell responses is due in part to the development of regulatory T cell activity that can suppress the activation of previously primed Th effector cells in a contact-dependent manner. Depletion of CD25+FoxP3+cells, and addition of exogenous IL-2 partially rescues Th1 responses in effector T cells co-cultured with ganglioside primed T cells. Collectively, these data suggest that ganglioside-exposed dendritic cells promote regulatory T cell activity that may have long lasting effects on the development of tumor specific immune responses.

Phosphatidylinositol 4-phosphate 5-kinase α is induced in ganglioside-stimulated brain astrocytes and contributes to inflammatory responses

In brain tissue, astrocytes play defensive roles in central nervous system integrity by mediating immune responses against pathological conditions. Type I phosphatidylinositol 4-phosphate 5-kinase alpha (PIP5K alpha) that is responsible for production of phosphatidylinositol 4,5-bisphosphate (PI[4,5]P2) regulates many important cell functions at the cell surface. Here, we have examined whether PIP5K alpha is associated with astrocyte inflammatory responses. Gangliosides are releasable from damaged cell membranes of neurons and capable of inducing inflammatory responses. We found that treatment of primary cultured astrocytes with gangliosides significantly enhanced PIP5K alpha mRNA and protein expression levels. PI(4,5)P2 imaging using a fluorescent tubby (R332H) expression as a PI(4,5)P2-specific probe showed that ganglioside treatment increased PI(4,5)P2 level. Interestingly, microRNA-based PIP5K alpha knockdown strongly reduced ganglioside-induced transcription of proinflammatory cytokines IL-1 beta and TNFalpha. PIP5K alpha knockdown also suppressed ganglioside-induced phosphorylation and nuclear translocation of NF-kappaB and the degradation of I kappaB-alpha, indicating that PIP5K alpha knockdown interfered with the ganglioside-activated NF-kappaB signaling. Together, these results suggest that PIP5K alpha is a novel inflammatory mediator that undergoes upregulation and contributes to immune responses by facilitating NF-kappaB activation in ganglioside-stimulated astrocytes.

The Behavioral and Immunological Effect of GM-1 Ganglioside on Nerve Root Regeneration Following C5 Nerve Root Avulsion in a Rat Model

Preganglionic nerve root avulsion precludes sensory return, but motor regeneration is possible with sparing of motoneurons. The effect of GM-1 ganglioside treatment was studied with parallel evaluation of the autoimmune response. Rats (N=64) received injections of either GM-1 ganglioside or saline for 30 days following either C5 root avulsion or a hemilaminectomy control. The Bertelli grooming test assessed functional return. Before sacrifice at 5 months, serum was collected for enzyme-linked immunoabsorbent assay testing. Only 44% of the rats treated with ganglioside had a good functional outcome compared with 50% for controls. Although 17% of the rats developed anti GM-1 antibodies, there was no functional or histological evidence of neuropathy in any of the rats. We conclude that ganglioside treatment did not enhance recovery from peripheral nerve injury. Although an immune response was present in some rats, no overt signs of neuropathy were observed.

Inhibition of TLR Activation and Up-Regulation of IL-1R-Associated Kinase-M Expression by Exogenous Gangliosides

Gangliosides, sialic acid-containing glycosphingolipids present in the outer leaflet of plasma membranes, are produced at high levels by some tumors, are actively shed into the tumor microenvironment, and can be detected in high concentrations in the serum of cancer patients. These tumor-shed molecules are known to be immunosuppressive, although mechanisms remain to be fully elucidated. In this study, we show that membrane enrichment of human monocytes with purified exogenous gangliosides potently inhibits ligand-induced activation and proinflammatory cytokine production induced by a broad range of TLRs, including TLR2, TLR3, TLR6, and TLR7/8, in addition to a previously identified inhibitory effect on TLR4 and TLR5. Inhibition of TLR activation is reversible, with complete restoration of TLR signaling within 6-24 h of washout of exogenous gangliosides, and is selective for certain gangliosides (GM1, GD1a, and GD1b), whereas others (GM3) are inactive. To characterize the inhibition, we assessed the expression of the TLR signaling pathway inhibitor, IL-1 receptor associated kinase-M (IRAK-M). In response to ganglioside enrichment alone, we observed striking up-regulation of IRAK-M in monocytes, but without concomitant proinflammatory cytokine production. This contrasts with endotoxin tolerance, in which IRAK-M up-regulation follows proinflammatory cytokine expression caused by LPS exposure. We hypothesize that ganglioside treatment induces a state of tolerance to TLR signaling, leading to blunted activation of innate immune responses. In the tumor microenvironment, shed tumor ganglioside enrichment of APC membranes may likewise cause these cells to bypass the normal TLR signaling response and progress directly to the inhibitory state.

Time Dependent Changes in Diabetic Cystopathy in Rats Include Compensated and Decompensated Bladder Function

Diabetic bladder dysfunction is among the most common and bothersome complications of diabetes mellitus. While bladder filling and voiding problems have been reported, the precise functional changes in diabetic bladders remain unclear. We investigated time dependent changes in bladder function in streptozotocin induced diabetic rats. Cystometrograms and detrusor muscle contractility were examined in male age matched control and diabetic Sprague-Dawley rats (Harlan, Indianapolis, Indiana) 3, 6, 9, 12 and 20 weeks after diabetes induction with streptozotocin. Diabetes decreased average body weight and increased bladder weight, capacity and compliance. Peak detrusor leak pressure increased gradually from weeks 3 to 6 to 9 in diabetic rats (mean +/- SEM 47.3 +/- 2.5, 50.8 +/- 3.0 and 56.0 +/- 3.6 cm H(2)O) and in controls (36.9 +/- 1.4, 37.7 +/- 1.5 and 41.6 +/- 1.81 cm H(2)O, respectively). However, at 12 and 20 weeks diabetic rats deviated strongly from this trend with peak detrusor leak pressure decreasing vs controls (41.6 +/- 2.8 and 37.3 +/- 0.9 vs 45.2 +/- 1.7 and 49.6 +/- 1.4 cm H(2)O, respectively) and post-void resting pressures increasing from 9-week levels vs controls (interactions p <0.0001). In contractility studies increased contractile force responses of diabetic animals to carbamylcholine chloride, potassium chloride, adenosine 5'-triphosphate and electric field stimulation peaked at 6 or 9 weeks but at 12 to 20 weeks they generally reverted toward those of controls (carbamylcholine chloride and electrical field stimulation interactions p = 0.0022 and 0.01, respectively). Diabetic bladders may undergo a transition from a compensated to a decompensated state and transition in the streptozotocin rat model may begin 9 to 12 weeks after induction.

Gangliosides for acute spinal cord injury

Spinal cord injury (SCI) results in loss of feeling and movement. The consequences can be devastating for the patient and his or her carers. Global estimates of the number of new cases annually range from 15 to 40 per million. Leading causes of acute SCI are road traffic injury, violence, and injuries sustained in sports and other recreational activities. Care for people with SCI has improved, leading to an increase in survival rates. Attempts to improve patients' feeling and movement have involved the use of a wide range of treatments. Gangliosides are compounds that occur naturally in cell membranes. Laboratory studies have suggested they may have protective effects on nerves and even help them to re-grow. Clinical trials have taken place using gangliosides (usually GM1 ganglioside) for a number of neurological conditions. To quantify the evidence for the effectiveness and safety of gangliosides when used to treat acute SCI. We searched the following databases to identify trials for inclusion: CENTRAL, MEDLINE, EMBASE, and the National Research Register. We also searched web-based trials registers, such as Current Controlled Trials. We approached the manufacturers of the most widely used ganglioside and researchers in this field to try to locate any unpublished data. Randomised controlled trials of any ganglioside versus controls, in patients with SCI. Outcome measures specified were: mortality, recovery of motor function, improvement in sensory measures, measures of functional activity, infections and any other adverse events. Data were extracted from published studies and authors were contacted for further information. All data found was dichotomous and odds ratios (with 95% CIs) were calculated. A fixed-effects model was assumed. Two studies met the inclusion criteria. There were no deaths in one (n=37). In the other (n=760), there were slightly more deaths in the treatment group than in the control group; odds ratio 1.07 (0.57, 2.00 95%CI) - a result that can be explained by the play of chance. Methodological weaknesses regarding the collection and presentation of data from the two studies made it impossible to reach any conclusions regarding the effect of gangliosides on the other specified outcomes. The evidence available does not support the use of ganglioside treatment to reduce the death rate in SCI patients. No evidence has yet emerged that ganglioside treatment improves recovery or quality of life in survivors.

Gangliosides activate microglia via protein kinase C and NADPH oxidase.

Microglia, the major immune effector cells in the central nervous system, are activated when the brain suffers injury. A number of studies indicate that gangliosides activate microglia. However, the signaling mechanisms involved in microglial activation are not yet to be elucidated. Our results show that gangliosides induce the expression of interleukin (IL)-1beta, tumor necrosis factor-alpha (TNF-alpha), and inducible nitric oxide synthase (iNOS) in rat brain microglia and BV2 murine microglia via protein kinase C (PKC) and NADPH oxidase. Expression of IL-1beta, TNF-alpha, and iNOS in ganglioside-treated cells was significantly reduced in the presence of inhibitors of PKC (GF109203X, Go6976, Ro31-8220, and rottlerin) and NADPH oxidase (diphenyleneiodonium chloride [DPI]). In response to gangliosides, PKC-alpha, betaII, and delta and NADPH oxidase p67(phox) translocated from the cytosol to the membrane. ROS generation was also activated within 5 min of ganglioside treatment. Ganglioside-induced ROS generation was blocked by PKC inhibitors. Furthermore, ganglioside-induced activation of NF-kappaB, an essential transcription factor that mediates the expression of IL-1beta, TNF-alpha, and iNOS, was reduced in the presence of GF109203X and DPI. Our results collectively suggest that gangliosides activate microglia via PKC and NADPH oxidase, which regulate activation of NF-kappaB.