Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Reduces Fibromyalgia Pain by Attenuating the HMGB1, S100B, and TRPV1 Signalling Pathways in the Mouse Brain

doi: 10.1155/2022/2242074

Figure Lengend Snippet: Comparative graph showing the hind paw withdrawal threshold and latency. Black: control group, red: ICS group, blue: ICS + electroacupuncture (EA) group, green: transient receptor vanilloid member one deletion ( Trpv1 −/− ) group. ∗ indicates statistical significance with p < 0.05 when compared with the control group. # indicates statistical significance with p < 0.05 when compared with the ICS group. (a) Mechanical hyperalgesia measured by the von Frey test in the four groups. (b) Thermal hyperalgesia assessed by the Hargreaves test in the four groups. (c) Schematic illustration showing the ICS procedure. Control mice stayed in a 24°C environment day and night for five days. Mice subjected to ICS were kept in a 4°C environment for 17.5 hours (from 4:30 p.m. to 10:00 a.m.) from Day 0 to 3. Between 10:00 a.m. and 4:30 p.m. (total duration: 6.5 hours), they were placed in a room at 24°C. In this 6.5-hour period, the mice were subjected to intermittent environment temperature change (24°C and 4°C) for 30 min each time. The procedure was terminated on Day 3 at 10:00 a.m., and fibromyalgia-like pain was assessed.

Article Snippet: The membrane was blocked with 5% nonfat milk in TBS-T buffer (10 mM Tris-HCl pH 7.5, 100 mM NaCl, 0.1% Tween-20) and then incubated with a primary antibody against TRPV1 (∼95 kDa, 1 : 1000; Alomone, Israel), HMGB1 (∼28 kDa, 1 : 1000; Alomone, Israel), S100B (∼10 kDa, 1 : 1000; Millipore, USA), TLR4 (∼35 kDa, 1 : 1000; Millipore, USA), RAGE (∼42 kDa, 1 : 1000; Millipore, USA), pPI3K (∼125 kDa, 1 : 1000; Millipore, USA), pAkt (∼60 kDa, 1 : 1000; Millipore, USA), pmTOR (∼60 kDa, 1 : 500; Millipore, USA), pERK1/2 (∼42–44 kDa, 1 : 1000; Millipore, USA), pp38 (∼42 kDa, 1 : 1000; Millipore, USA), pJNK (∼42 kDa, 1 : 1000; Millipore, USA), and pNF- κ B (∼65 kDa, 1 : 1000; Millipore, USA) in TBS-T with 1% bovine serum albumin.

Techniques:

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Reduces Fibromyalgia Pain by Attenuating the HMGB1, S100B, and TRPV1 Signalling Pathways in the Mouse Brain

doi: 10.1155/2022/2242074

Figure Lengend Snippet: (a) Expression of extracellular neurotransmitters ((A) HMGB1, (B) S100B), receptors ((C) RAGE, (D) TLR2, (E) TLR4), and cytoplasmic inflammation signal molecules ((F) pPI3K, (G) pAkt, (H) pmTOR, (I) pERK, (J) pp38, (K) pJNK, (L) pNF- κ B) in the mouse PFC. The four lanes on the immunoblots correspond to the protein bands of, in order, control (Con), ICS-induced fibromyalgia (ICS), electroacupuncture (EA), and Trpv1 −/− groups. ∗ indicates statistical significance with P < 0.05 when compared with the Con group. # indicates statistical significance with P < 0.05 when compared with the ICS group. (b) Immunofluorescence staining of TLR2 and pNF- κ B in the mouse PFC. Immunopositive signals (light green, indicated by a red arrow) were detected for TLR2 and pNF- κ B. Scale bar: 100 μ m.

Article Snippet: The membrane was blocked with 5% nonfat milk in TBS-T buffer (10 mM Tris-HCl pH 7.5, 100 mM NaCl, 0.1% Tween-20) and then incubated with a primary antibody against TRPV1 (∼95 kDa, 1 : 1000; Alomone, Israel), HMGB1 (∼28 kDa, 1 : 1000; Alomone, Israel), S100B (∼10 kDa, 1 : 1000; Millipore, USA), TLR4 (∼35 kDa, 1 : 1000; Millipore, USA), RAGE (∼42 kDa, 1 : 1000; Millipore, USA), pPI3K (∼125 kDa, 1 : 1000; Millipore, USA), pAkt (∼60 kDa, 1 : 1000; Millipore, USA), pmTOR (∼60 kDa, 1 : 500; Millipore, USA), pERK1/2 (∼42–44 kDa, 1 : 1000; Millipore, USA), pp38 (∼42 kDa, 1 : 1000; Millipore, USA), pJNK (∼42 kDa, 1 : 1000; Millipore, USA), and pNF- κ B (∼65 kDa, 1 : 1000; Millipore, USA) in TBS-T with 1% bovine serum albumin.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Reduces Fibromyalgia Pain by Attenuating the HMGB1, S100B, and TRPV1 Signalling Pathways in the Mouse Brain

doi: 10.1155/2022/2242074

Figure Lengend Snippet: (a) Expression of extracellular neurotransmitters ((A) HMGB1, (B) S100B), receptors ((C) RAGE, (D) TLR2, (E) TLR4), and cytoplasmic inflammation signal molecules ((F) pPI3K, (G) pAkt, (H) pmTOR, (I) pERK, (J) pp38, (K) pJNK, (L) pNF- κ B) in the mouse SSC. The four lanes on the immunoblots correspond to the protein bands of, in order, control (Con), ICS-induced fibromyalgia (ICS), electroacupuncture (EA), and Trpv1 −/− groups. ∗ indicates statistical significance with P < 0.05 when compared with the Con group. # indicates P < 0.05 statistical significance when compared with the ICS group. (b) Immunofluorescence staining of TLR2 and pNF- κ B in the mouse SSC. Immunopositive signals (light green, indicated by a red arrow) were detected for TLR2 and pNF- κ B. Scale bar: 100 μ m.

Article Snippet: The membrane was blocked with 5% nonfat milk in TBS-T buffer (10 mM Tris-HCl pH 7.5, 100 mM NaCl, 0.1% Tween-20) and then incubated with a primary antibody against TRPV1 (∼95 kDa, 1 : 1000; Alomone, Israel), HMGB1 (∼28 kDa, 1 : 1000; Alomone, Israel), S100B (∼10 kDa, 1 : 1000; Millipore, USA), TLR4 (∼35 kDa, 1 : 1000; Millipore, USA), RAGE (∼42 kDa, 1 : 1000; Millipore, USA), pPI3K (∼125 kDa, 1 : 1000; Millipore, USA), pAkt (∼60 kDa, 1 : 1000; Millipore, USA), pmTOR (∼60 kDa, 1 : 500; Millipore, USA), pERK1/2 (∼42–44 kDa, 1 : 1000; Millipore, USA), pp38 (∼42 kDa, 1 : 1000; Millipore, USA), pJNK (∼42 kDa, 1 : 1000; Millipore, USA), and pNF- κ B (∼65 kDa, 1 : 1000; Millipore, USA) in TBS-T with 1% bovine serum albumin.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Reduces Fibromyalgia Pain by Attenuating the HMGB1, S100B, and TRPV1 Signalling Pathways in the Mouse Brain

doi: 10.1155/2022/2242074

Figure Lengend Snippet: (a) Expression of extracellular neurotransmitters (A) HMGB1, (B) S100B), receptors (C) RAGE, (D) TLR2, (E) TLR4) and cytoplasmic inflammation signal molecules (F) pPI3K, (G) pAkt, (H) pmTOR, (I) pERK, (J) pp38, (K) pJNK, (L) pNF- κ B) in the mouse thalamus. The four lanes on the immunoblots correspond to the protein bands of, in order, control (Con), ICS-induced fibromyalgia (ICS), electroacupuncture (EA) and Trpv1 −/− groups. ∗ indicates statistical significance with P < 0.05 when compared with the Con group. # indicates statistical significance with P < 0.05 when compared to the ICS group. (b) Immunofluorescence staining of TLR2 and pNF- κ B in the mouse thalamus. Immunopositive signals (light green, indicated by a red arrow) were detected for TLR2 and pNF- κ B. Scale bar: 100 μ m.

Article Snippet: The membrane was blocked with 5% nonfat milk in TBS-T buffer (10 mM Tris-HCl pH 7.5, 100 mM NaCl, 0.1% Tween-20) and then incubated with a primary antibody against TRPV1 (∼95 kDa, 1 : 1000; Alomone, Israel), HMGB1 (∼28 kDa, 1 : 1000; Alomone, Israel), S100B (∼10 kDa, 1 : 1000; Millipore, USA), TLR4 (∼35 kDa, 1 : 1000; Millipore, USA), RAGE (∼42 kDa, 1 : 1000; Millipore, USA), pPI3K (∼125 kDa, 1 : 1000; Millipore, USA), pAkt (∼60 kDa, 1 : 1000; Millipore, USA), pmTOR (∼60 kDa, 1 : 500; Millipore, USA), pERK1/2 (∼42–44 kDa, 1 : 1000; Millipore, USA), pp38 (∼42 kDa, 1 : 1000; Millipore, USA), pJNK (∼42 kDa, 1 : 1000; Millipore, USA), and pNF- κ B (∼65 kDa, 1 : 1000; Millipore, USA) in TBS-T with 1% bovine serum albumin.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Reduces Fibromyalgia Pain by Attenuating the HMGB1, S100B, and TRPV1 Signalling Pathways in the Mouse Brain

doi: 10.1155/2022/2242074

Figure Lengend Snippet: (a) The expression of extracellular neurotransmitters ((A) HMGB1, (B) S100B), receptors ((C) RAGE, (D) TLR2, (E) TLR4), and cytoplasmic inflammation signal molecules ((F) pPI3K, (G) pAkt, (H) pmTOR, (I) pERK, (J) pp38, (K) pJNK, (L) pNF- κ B) in the mouse amygdala. The four lanes on the immunoblots correspond to the protein bands of, in order, control (Con), ICS-induced fibromyalgia (ICS), electroacupuncture (EA), and Trpv1 −/− groups. ∗ indicates statistical significance with P < 0.05 when compared with the Con group. # indicates statistical significance with P < 0.05 when compared with the ICS group. (b) Immunofluorescence staining of TLR2 and pNF- κ B in the mouse amygdala. Immunopositive signals (light green, indicated by a red arrow) were detected for TLR2 and pNF- κ B. Scale bar: 100 μ m.

Article Snippet: The membrane was blocked with 5% nonfat milk in TBS-T buffer (10 mM Tris-HCl pH 7.5, 100 mM NaCl, 0.1% Tween-20) and then incubated with a primary antibody against TRPV1 (∼95 kDa, 1 : 1000; Alomone, Israel), HMGB1 (∼28 kDa, 1 : 1000; Alomone, Israel), S100B (∼10 kDa, 1 : 1000; Millipore, USA), TLR4 (∼35 kDa, 1 : 1000; Millipore, USA), RAGE (∼42 kDa, 1 : 1000; Millipore, USA), pPI3K (∼125 kDa, 1 : 1000; Millipore, USA), pAkt (∼60 kDa, 1 : 1000; Millipore, USA), pmTOR (∼60 kDa, 1 : 500; Millipore, USA), pERK1/2 (∼42–44 kDa, 1 : 1000; Millipore, USA), pp38 (∼42 kDa, 1 : 1000; Millipore, USA), pJNK (∼42 kDa, 1 : 1000; Millipore, USA), and pNF- κ B (∼65 kDa, 1 : 1000; Millipore, USA) in TBS-T with 1% bovine serum albumin.

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Electroacupuncture Reduces Fibromyalgia Pain by Attenuating the HMGB1, S100B, and TRPV1 Signalling Pathways in the Mouse Brain

doi: 10.1155/2022/2242074

Figure Lengend Snippet: Schematic illustration of neuronal and non-neuronal mechanisms underlying EA-mediated analgesic effect on ICS-induced fibromyalgia pain. The summary diagram shows the importance of and mechanisms involving glial cells (astrocytes and microglia) and TRPV1 in fibromyalgia pain. EA inhibits HMGB1 and S100B release from non-neuronal cells or directly inhibits TRPV1 on the plasma membrane. Mice with a TRPV1 gene deletion ( Trpv1 −/− ) have the same phenotype than mice treated with EA.

Article Snippet: The membrane was blocked with 5% nonfat milk in TBS-T buffer (10 mM Tris-HCl pH 7.5, 100 mM NaCl, 0.1% Tween-20) and then incubated with a primary antibody against TRPV1 (∼95 kDa, 1 : 1000; Alomone, Israel), HMGB1 (∼28 kDa, 1 : 1000; Alomone, Israel), S100B (∼10 kDa, 1 : 1000; Millipore, USA), TLR4 (∼35 kDa, 1 : 1000; Millipore, USA), RAGE (∼42 kDa, 1 : 1000; Millipore, USA), pPI3K (∼125 kDa, 1 : 1000; Millipore, USA), pAkt (∼60 kDa, 1 : 1000; Millipore, USA), pmTOR (∼60 kDa, 1 : 500; Millipore, USA), pERK1/2 (∼42–44 kDa, 1 : 1000; Millipore, USA), pp38 (∼42 kDa, 1 : 1000; Millipore, USA), pJNK (∼42 kDa, 1 : 1000; Millipore, USA), and pNF- κ B (∼65 kDa, 1 : 1000; Millipore, USA) in TBS-T with 1% bovine serum albumin.

Techniques: