Bee Venom Acupuncture Alleviates Experimental Autoimmune Encephalomyelitis by Upregulating Regulatory T Cells and Suppressing Th1 and Th17 Responses

Abstract

The protective and therapeutic mechanism of bee venom acupuncture (BVA) in neurodegenerative disorders is not clear. We investigated whether treatment with BVA (0.25 and 0.8 mg/kg) at the Zusanli (ST36) acupoints, located lateral from the anterior border of the tibia, has a beneficial effect in a myelin basic protein (MBP)(68-82)-induced acute experimental autoimmune encephalomyelitis (EAE) rat model. Pretreatment (every 3 days from 1 h before immunization) with BVA was more effective than posttreatment (daily after immunization) with BVA with respect to clinical signs (neurological impairment and loss of body weight) of acute EAE rats. Treatment with BVA at the ST36 acupoint in normal rats did not induce the clinical signs. Pretreatment with BVA suppressed demyelination, glial activation, expression of cytokines [interferon (IFN)-γ, IL-17, IL-17A, tumor necrosis factor-alpha (TNF-α), and IL-1β], chemokines [RANTES, monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein (MIP)-1α], and inducible nitric oxide synthase (iNOS), and activation of p38 mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-κB (p65 and phospho-IκBα) signaling pathways in the spinal cord of acute EAE rats. Pretreatment with BVA decreased the number of CD4(+), CD4(+)/IFN-γ(+), and CD4(+)/IL-17(+) T cells, but increased the number of CD4(+)/Foxp3(+) T cells in the spinal cord and lymph nodes of acute EAE rats. Treatment with BVA at six placebo acupoints (SP9, GB39, and four non-acupoints) did not have a positive effect in acute EAE rats. Interestingly, onset and posttreatment with BVA at the ST36 acupoint markedly attenuated neurological impairment in myelin oligodendrocyte glycoprotein (MOG)(35-55)-induced chronic EAE mice compared to treatment with BVA at six placebo acupoints. Our findings strongly suggest that treatment with BVA with ST36 acupoint could delay or attenuate the development and progression of EAE by upregulating regulatory T cells and suppressing T-helper (Th) 17 and Th1 responses. These results warrant further investigation of BVA as a treatment for autoimmune disorders of the central nervous system.

Sulforaphane Ameliorates 3-Nitropropionic Acid-Induced Striatal Toxicity by Activating the Keap1-Nrf2-ARE Pathway and Inhibiting the MAPKs and NF-κB Pathways

“…Immunohistochemical analysis was performed as previously described [9,10,26,27]. Briefly, free floating brain sections (30-μm thickness) from each group (sham, n=3; 3-NP, n=5; 3-NP+SFN 5.0, n=5, and SFN 5.0, n=3) were incubated for 30 min with 3 % H 2 O 2 in PBS, and washed in PBS, and then blocked with a solution containing 5 % normal goat/or horse serum, 2 % fetal bovine serum, 2 % bovine serum albumin, and 0.1 % Triton X-100 for 2 h at RT.…”

“…Western blot analysis was performed as previously described [9,10,26,27] Fig. 1 Experimental protocols used for treatment with 3-NP, SFN, and activators of Nrf2.…”

“…To investigate the mRNA level of pro-inflammatory cytokines, chemokines, and neuromodulators, real-time PCR analysis was performed as previously described [9,10,[26][27][28]. Briefly, 12 h after the last injection of 3-NP, the mice (sham, n=3; 3-NP, n=5; 3-NP+SFN 5.0, n=5; SFN, n=3) were anesthetized, and each striatum was removed and deepfrozen.…”

Oriental Medicine Woohwangchungsimwon Attenuates Kainic Acid-Induced Seizures and Neuronal Cell Death in the Hippocampus

“…The immunohistochemistry was carried out as previously described [22][23][24] using rabbit anti-ionized calcium binding adaptor molecule-1 (Iba-1) (1:2000; Wako) or rabbit-active (cleaved) caspase-3 antibody (1:1000; Cell Signaling Technology) antibodies. A semiquantitative histological evaluation for Iba-1 antibody (40· magnified microscope field) was conducted and scored blindly by two observers using the following scale: resting microglia, mildly activated microglia, and moderately activated microglia.…”

“…The primer sequences of mouse interleukin (IL)-1b, IL-6, tumor necrosis factor (TNF)-a, cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were previously described. 22,23 Expression levels of each gene were normalized to that of GAPDH.…”