Journal:

Article Title: Cannabinoids Ameliorate Pain and Reduce Disease Pathology in Cerulein-Induced Acute Pancreatitis

doi: 10.1053/j.gastro.2007.02.035

Figure Lengend Snippet: Induction of the endocannabinoid system in acute pancreatitis. Immunohistochemical detection of CB1 and CB2 is shown in human pancreas (A–C and G–I) or mouse pancreas (D–F and J–L) derived either from control subjects (A, D, G, and J) or subjects with acute pancreatitis (B and C, E and F, H and I, K and L). (C, F, I and L) Areas of necrosis within human and mouse acute pancreatitis are indicated by dotted lines. In normal human pancreas (A) CB1-immunoreactivity in acinar cells (▶; magnified in inset C), ducts (→), and nerves (magnified in insets), was weak but strong in acute pancreatitis (B, C). (D–F) Similarly, pancreas demonstrated increased immunoreactivity for CB1 after induction of acute pancreatitis with cerulein (E, F) over normal mouse pancreas (D). Intense CB1 immunostaining is observed on acinar cell necrosis (dotted arrow; F). (G–I) Moderate staining for CB2 is seen in normal human pancreas (G) with a slight increase in acute pancreatitis (H, I) in acinar cells (▶; magnified in inset I), ducts (→), and nerves (magnified in insets). Upon induction of acute pancreatitis in mice, there is a pronounced increase in CB2 immunoreactivity over expression levels in normal mouse pancreas (J–L). This is particularly present within acinar cell necrosis (dotted arrow; L). Original magnification: 40x (A and B, D and E, G and H, J and K) or 80x (F and L; insets in A–C and G–I) objective. (M, N) Immunoblot analysis of pancreas samples derived from acute pancreatitis (lanes 1–3) and control human donors (lanes 4–6) with antibodies recognizing CB1 (M) or CB2 (N). (▶) indicates anti-CB1 (at approximately 128 kDa) and anti-CB2-immunoreactive bands (at 38 and 26 kDa), which were abolished by preadsorption with the respective blocking peptide (lower lanes). Anti-ERK2 was used as an equal loading control. (O) In humans, pancreatic concentrations of the endocannabinoid anandamide (AEA) are higher in acute pancreatitis (black bars) than in normal pancreas (white bars; P < .05), whereas levels of combined 1- plus 2-arachidonoylglycerol (1 - AG + 2 - AG) are unchanged.

Article Snippet: Immunohistochemistry on paraffin sections of human pancreas was performed using rabbit anti-CB1 and anti-CB2 antibodies (Cayman Chemical, Ann Arbor, MI) as described previously.

Techniques: Immunohistochemical staining, Derivative Assay, Control, Immunostaining, Staining, Over Expression, Western Blot, Blocking Assay

Journal:

Article Title: Cannabinoids Ameliorate Pain and Reduce Disease Pathology in Cerulein-Induced Acute Pancreatitis

doi: 10.1053/j.gastro.2007.02.035

Figure Lengend Snippet: Levels of serological, molecular, and inflammatory markers in acute pancreatitis and effects of CB1/CB2 antagonists. (A) Levels of serum lipase, pancreatic MPO, and the pancreatic wet weight/dry weight ratio are increased in mice following cerulein-induced pancreatitis (black bars) over control animals (white bars; * P < .05). (B) Pancreatic levels of IL-6 protein and KC1 mRNA (normalized to GAPDH expression levels) rise significantly following induction of acute pancreatitis. Treatment with cannabinoid receptor antagonists AM251 or AM630 either alone or in combination did not change levels of these parameters, except pancreatic KC1 mRNA expression (P < .05). * and # represent P < .05 as compared with the saline group or the cerulein group, respectively. (*) Mann–Whitney U test. (#) Analysis of variance followed by post hoc Bonferroni’s multiple comparison test. n.s. = statistically not significant.

Article Snippet: Immunohistochemistry on paraffin sections of human pancreas was performed using rabbit anti-CB1 and anti-CB2 antibodies (Cayman Chemical, Ann Arbor, MI) as described previously.

Techniques: Control, Expressing, Saline, MANN-WHITNEY, Comparison

Journal: International Journal of Molecular Sciences

Article Title: CB2 Receptor Activation Inhibits Melanoma Cell Transmigration through the Blood-Brain Barrier

doi: 10.3390/ijms15058063

Figure Lengend Snippet: Expression of cannabinoid and cannabinoid-like receptors in brain endothelial and melanoma cells. RT-PCR was performed to determine the expression of CB2A and CB2B (positive control: HL-60) transcriptional variants of human CB2 receptor in hCMEC/D3 brain endothelial and A2058 melanoma cells ( a , b ); the expression of transcriptional variant 1 and 2 of rat CB2 receptor ( c , d ) in rat brain endothelial cells (RBECs) (positive control: rat spleen), the expression of CB1 receptor ( e ); transcriptional variant 1 and 2 of GPR18 ( f , g ); GPR119 ( h ) and GPR55 ( i ) in hCMEC/D3 human brain endothelial cells and A2058 melanoma cells. Dotted arrows indicate the absence of specific bands.

Article Snippet: The selective CB2 agonist JWH-133 solution (diluted in Tocrisolve) was purchased from Tocris.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Positive Control, Variant Assay

Journal: International Journal of Molecular Sciences

Article Title: CB2 Receptor Activation Inhibits Melanoma Cell Transmigration through the Blood-Brain Barrier

doi: 10.3390/ijms15058063

Figure Lengend Snippet: Effect of CB2 activation on the attachment of melanoma cells on the brain endothelium. Results are represented as % control ( i.e. , D3 + A2058) and given as mean ± SD. N = 3. * p < 0.05 as assessed by ANOVA and Bonferroni’s post-hoc test. ( a ) D3(jwh-133) and A2058(jwh-133) represent cells pre-treated with 10 μM JWH-133 for 4 h. D3 + A2058 + JWH-133 denotes cells treated with 10 μM JWH-133 during the 90 min adhesion assay; ( b ) JWH-133 (10 μM), U0126 (10 μM) and PTX (100 ng/mL) were applied in pre-treatment of both cell types and treatment during the adhesion assay.

Article Snippet: The selective CB2 agonist JWH-133 solution (diluted in Tocrisolve) was purchased from Tocris.

Techniques: Activation Assay, Control, Cell Adhesion Assay

Journal: International Journal of Molecular Sciences

Article Title: CB2 Receptor Activation Inhibits Melanoma Cell Transmigration through the Blood-Brain Barrier

doi: 10.3390/ijms15058063

Figure Lengend Snippet: Effect of CB2 activation on the transendothelial migration of melanoma cells. Results are represented as % control ( i.e. , RBEC + A2058) and given as mean ± SD. N = 3. * p < 0.01 (compared to control) as assessed by ANOVA and Bonferroni’s post-hoc test. ( a ) RBEC(jwh-133) represents endothelial cells pre-treated with 10 μM JWH-133 for 4 h. RBEC(jwh-133) + A2058(jwh-133) + JWH-133 denotes that both endothelial and melanoma cells were pre-treated with 10 μM JWH-133 for 4 h and treated with 10 μM JWH-133 during the 5 h transmigration assay; ( b ) JWH-133 (10 μM) and SR-144528 (10 μM) were applied in pre-treatment of both cell types and treatment during the transmigration assay.

Article Snippet: The selective CB2 agonist JWH-133 solution (diluted in Tocrisolve) was purchased from Tocris.

Techniques: Activation Assay, Migration, Control, Transmigration Assay

Journal: Neural Regeneration Research

Article Title: Electroacupuncture exerts neuroprotective effects on ischemia/reperfusion injury in JNK knockout mice: the underlying mechanism

doi: 10.4103/1673-5374.235294

Figure Lengend Snippet: Effects of Electroacupuncture on the neurological deficit of the JNK knockout mice with I/R injury. Data are presented as the mean ± SD ( n = 6); one-way analysis of variance followed by least significant difference post hoc test was used. * P < 0.05, vs . I/R group; # P < 0.05, vs . sham group. EA: Electroacupuncture; I/R: ischemia/reperfusion; JNK: c-Jun N-terminal kinase.

Article Snippet: A total of 54 JNK knockout mice (half males and half females), weighing 20–25 g, were purchased from Cyagen Biosciences Co. Ltd. (Guangzhou, China; license number: SCXK (Yue) 2013-0002).

Techniques: Knock-Out

Journal: Neural Regeneration Research

Article Title: Electroacupuncture exerts neuroprotective effects on ischemia/reperfusion injury in JNK knockout mice: the underlying mechanism

doi: 10.4103/1673-5374.235294

Figure Lengend Snippet: The p-ERK and p-p38 immunoreactivity in the hippocampus and cortex of JNK knockout mice with cerebral I/R injury. (A, D) The p-ERK immunoreactivity in the hippocampus and cortex of JNK knockout mice. (B, E) The p-p38 immunoreactivity in the hippocampus and cortex of JNK knockout mice. Data are representative of six individual mice in each group. Data are expressed as the mean ± SD ( n = 6); one-way analysis of variance followed by least significant difference post hoc test. * P < 0.05, vs . I/R group; # P < 0.05, vs . sham group. (C) Expression of p-ERK and p-p38 in the hippocampus of JNK knockout mice at 3 days under a laser scanning confocal microscope (scale bars: 9 μm). (F) Expression of p-ERK and p-p38 in the cortex of JNK knockout mice at 3 days under a laser scanning confocal microscope (original magnification, 400×). Arrows indicate p-ERK and p-p38 immunoreactivity. EA: Electroacupuncture; I/R: ischemia/reperfusion; JNK: c-Jun N-terminal kinase; p-ERK: phospho-extracellular signal-regulated kinase; DAPI: 4′,6-diamidino-2-phenylindole; IOD: integrated optical density.

Article Snippet: A total of 54 JNK knockout mice (half males and half females), weighing 20–25 g, were purchased from Cyagen Biosciences Co. Ltd. (Guangzhou, China; license number: SCXK (Yue) 2013-0002).

Techniques: Knock-Out, Expressing, Microscopy

Journal: Neural Regeneration Research

Article Title: Electroacupuncture exerts neuroprotective effects on ischemia/reperfusion injury in JNK knockout mice: the underlying mechanism

doi: 10.4103/1673-5374.235294

Figure Lengend Snippet: The mRNA expression of ERK and p38 in the injured brain tissues of JNK knockout mice with cerebral I/R injury. (A, B) The mRNA expression of ERK (A) and p38 (B) in each group at different time points was assessed by quantitative real-time polymerase chain reaction. Data are presented as the mean ± SD ( n = 3); one-way analysis of variance followed by least significant difference post hoc test was used. * P < 0.05, vs . I/R group; # P < 0.05, vs . sham group. EA: Electroacupuncture; I/R: ischemia/reperfusion; ERK: extracellular signal-regulated kinase.

Article Snippet: A total of 54 JNK knockout mice (half males and half females), weighing 20–25 g, were purchased from Cyagen Biosciences Co. Ltd. (Guangzhou, China; license number: SCXK (Yue) 2013-0002).

Techniques: Expressing, Knock-Out, Real-time Polymerase Chain Reaction

Journal: Neural Regeneration Research

Article Title: Electroacupuncture exerts neuroprotective effects on ischemia/reperfusion injury in JNK knockout mice: the underlying mechanism

doi: 10.4103/1673-5374.235294

Figure Lengend Snippet: Protein expression of p-ERK and p-P38 in the JNK knockout mice with I/R injury. (A) Relative expression of p-ERK in each group at 3 days. (B) Relative expression p-p38 in each group at 3 days. Data are presented as the mean ± SD ( n = 3); one-way analysis of variance followed by least significant difference post hoc test was used. * P < 0.05, vs . I/R group; # P < 0.05, vs . sham group. (C) Bands of p-ERK and p-p38 assessed by western blot assay. EA: Electroacupuncture; I/R: ischemia/reperfusion; JNK: c-Jun N-terminal kinase; ERK: extracellular signal-regulated kinase; p-ERK: phospho-extracellular signal-regulated kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: A total of 54 JNK knockout mice (half males and half females), weighing 20–25 g, were purchased from Cyagen Biosciences Co. Ltd. (Guangzhou, China; license number: SCXK (Yue) 2013-0002).

Techniques: Expressing, Knock-Out, Western Blot