Journal: Pharmaceutical Biology

Article Title: Veratramine ameliorates pain symptoms in rats with diabetic peripheral neuropathy by inhibiting activation of the SIGMAR1-NMDAR pathway

doi: 10.1080/13880209.2022.2136207

Figure Lengend Snippet: Effect of veratramine on pain parameters of diabetic peripheral neuropathy. (A) Fasting blood glucose values in rats; (B) Mechanical withdrawal threshold (MWT) in rats before veratramine administration; (C) FBG detection results of each group at the end of the experiment. (D) MWT in rats after veratramine administration; (E) Plantar thermal paw withdrawal latency in rats after veratramine administration; (F) Hind paw cold allodynia. ** p < 0.01, * p < 0.05, vs. normal or vehicle group, ## p < 0.01, # p < 0.05, treated group vs. model group.

Article Snippet: The model group (model, n = 12) was given a high-fat diet (60% basic diet, 20% sucrose, 15% oil, and 5% cholesterol), and the treated group (treated, n = 12) was injected with 50 μg/kg veratramine (MedChemExpress Reagents Inc.

Techniques:

Journal: Pharmaceutical Biology

Article Title: Veratramine ameliorates pain symptoms in rats with diabetic peripheral neuropathy by inhibiting activation of the SIGMAR1-NMDAR pathway

doi: 10.1080/13880209.2022.2136207

Figure Lengend Snippet: Pathological changes of peripheral neuropathy in aged diabetic rats treated with veratramine. ( A) H&E photomicrographs of spinal cord tissue; (B) H&E photomicrographs of sciatic nerve. The arrows in the figure show that nerve cells show mitoses, and green circles represent nerve fibre enlargement or atrophy. The scale bar is 50 μm.

Article Snippet: The model group (model, n = 12) was given a high-fat diet (60% basic diet, 20% sucrose, 15% oil, and 5% cholesterol), and the treated group (treated, n = 12) was injected with 50 μg/kg veratramine (MedChemExpress Reagents Inc.

Techniques:

Journal: Pharmaceutical Biology

Article Title: Veratramine ameliorates pain symptoms in rats with diabetic peripheral neuropathy by inhibiting activation of the SIGMAR1-NMDAR pathway

doi: 10.1080/13880209.2022.2136207

Figure Lengend Snippet: Bioinformatics screening of veratramine target proteins. (A) Veratramine was visualised by docking with SHH protein, and its docking binding energy was –12.8 kcal/mol; (B) Veratramine was visualised by docking with SIGMAR1 protein, and its docking binding energy was –13.6 kcal/mol; (C) qPCR was used to detect SHH gene expression levels in the spinal cord tissues of rats in each group; (D) The spinal cord tissues of rats in each group were examined for SIGMAR1 gene expression levels by qPCR. ** p < 0.01, vs. vehicle group, ## p < 0.01, vs. model group.

Article Snippet: The model group (model, n = 12) was given a high-fat diet (60% basic diet, 20% sucrose, 15% oil, and 5% cholesterol), and the treated group (treated, n = 12) was injected with 50 μg/kg veratramine (MedChemExpress Reagents Inc.

Techniques: Binding Assay, Gene Expression

Journal: Pharmaceutical Biology

Article Title: Veratramine ameliorates pain symptoms in rats with diabetic peripheral neuropathy by inhibiting activation of the SIGMAR1-NMDAR pathway

doi: 10.1080/13880209.2022.2136207

Figure Lengend Snippet: Effect of veratramine on SIGMAR1 protein expression in spinal cord tissue of diabetic rats. (A) Immunohistochemistry (IHC) was used to stain and localise SIGMAR1 protein in rat spinal cord tissue, and the figure on the right is a statistical quantification figure of IHC results. (B) WB was used to detect the protein expression level of SIGMAR1. (C) Statistical quantification results of grey values of WB bands. ** p < 0.01, * p < 0.01, vs. vehicle group; ## p < 0.01, vs. model group. The scale bar is 50 μm.

Article Snippet: The model group (model, n = 12) was given a high-fat diet (60% basic diet, 20% sucrose, 15% oil, and 5% cholesterol), and the treated group (treated, n = 12) was injected with 50 μg/kg veratramine (MedChemExpress Reagents Inc.

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Pharmaceutical Biology

Article Title: Veratramine ameliorates pain symptoms in rats with diabetic peripheral neuropathy by inhibiting activation of the SIGMAR1-NMDAR pathway

doi: 10.1080/13880209.2022.2136207

Figure Lengend Snippet: Effect of veratramine on SIGMAR1 and its downstream regulatory proteins. (A) Coimmunoprecipitation (Co-IP) results of SIGMAR1 with NMDAR protein. (B) Statistical quantification map of band grey values in result A; (C) Co-IP results of CaMKII with NMDAR protein. (D) Statistical quantification map of band grey values in results C; (E) WB was used to detect the expression levels of NMDAR and its phosphorylated proteins. (F) Statistical quantification plot of grey values of WB bands. ** p < 0.01, * p < 0.01, vs. vehicle group; ## p < 0.01, vs. model group.

Article Snippet: The model group (model, n = 12) was given a high-fat diet (60% basic diet, 20% sucrose, 15% oil, and 5% cholesterol), and the treated group (treated, n = 12) was injected with 50 μg/kg veratramine (MedChemExpress Reagents Inc.

Techniques: Co-Immunoprecipitation Assay, Expressing

Journal: Pharmaceutical Biology

Article Title: Veratramine ameliorates pain symptoms in rats with diabetic peripheral neuropathy by inhibiting activation of the SIGMAR1-NMDAR pathway

doi: 10.1080/13880209.2022.2136207

Figure Lengend Snippet: Mechanism of action of veratramine on the SIGMAR1-NMDAR pathway. A mechanistic diagram of the SIGMAR1-NMDAR pathway in the pathologically induced DPN state. (A) SIGMAR1 can stabilise the intracellular Ca 2+ -CAMKII structure. (B) The stable Ca 2+ -CaMKII complex structure leads to decreased CaMKII in the free state. The inhibitory effect of CaMKII protein on NMDAR is weakened, and the activity of NMDAR increases. (C) After NMDAR activity is increased, extracellular Ca 2+ flows into the cell. (D) Increased intracellular Ca 2+ concentration consolidates the SIGMAR1-NMDAR complex structure and further enhances NMDAR activity. (B) Effects of veratramine treatment on the SIGMAR1-NMDAR pathway: After SIGMAR1 activity is inhibited by veratramine, the stability of the Ca 2+ -CaMKII complex structure is decreased, and the free CaMKII protein is increased, thus inhibiting NMDAR activity. When NMDAR activity is decreased, Ca 2+ inflow into cells decreases, and the stability of the SIGMAR1-NMDAR complex structure decreases, further weakening NMDAR protein activity.

Article Snippet: The model group (model, n = 12) was given a high-fat diet (60% basic diet, 20% sucrose, 15% oil, and 5% cholesterol), and the treated group (treated, n = 12) was injected with 50 μg/kg veratramine (MedChemExpress Reagents Inc.

Techniques: Activity Assay, Concentration Assay

Journal: Archives of biochemistry and biophysics

Article Title: A novel NADP(+)-dependent dehydrogenase activity for 7alpha/beta- and 11beta-hydroxysteroids in human liver nuclei: A third 11beta-hydroxysteroid dehydrogenase.

doi: 10.1016/j.abb.2009.04.010

Figure Lengend Snippet: Fig. 1. Concentration dependence of oxidative and reductive metabolism of 11b- HSD substrates by human liver microsomal fractions. Human liver protein (0.5 mg/ mL) was incubated for 120 min with either 1 mM b-NADPH min regenerating system or 1 mM b-NADP+ as described in the method section. (A) Oxidation of CS by b-NADP+ [4] or reduction of DHC by b-NADPH [s]. (B) Oxidation of 7a-OH-DHEA by b-NADP+ [4] or reduction of 7-oxo-DHEA by b-NADPH [s]. The rate of metabolism was expressed nmol product formed per mg protein per minute (mean value ± SEM; n = 5).

Article Snippet: DHEA, 7a-OH-DHEA, 7b-OY-DHEA and 7-oxo-DHEA, 11a-hydroxy-progesterone (11b-OH-PRO), 11b-hydroxy-progesterone (11b-OH-PRO), CS and DHC were obtained from Steraloids Inc. (Newport, RI).

Techniques: Concentration Assay, Incubation

Journal: Archives of biochemistry and biophysics

Article Title: A novel NADP(+)-dependent dehydrogenase activity for 7alpha/beta- and 11beta-hydroxysteroids in human liver nuclei: A third 11beta-hydroxysteroid dehydrogenase.

doi: 10.1016/j.abb.2009.04.010

Figure Lengend Snippet: Fig. 4. The effects of inhibitors of 11b-HSD on the metabolism of corticosterone and dehydrocorticosterone by human liver microsomal fractions. Liver microsomal fractions from human liver were incubated with either 1 mM b-NADPH and 10 lM DHC or 1 mM b-NADP+ and 10 lM CS for 120 min in the absence or presence of CBX (2 lM) or 50 lM of either 11a-hydroxy-progesterone, 11b-hydroxy-progesterone, dehydrocorticosterone, 7-oxo-DHEA or 7a-OH-DHEA as described in the method section. (A) Inhibition of the reduction of DHC by NADPH. (B) Inhibition of the oxidation of CS by NADP+. The rates were expressed as nmol CS or DHC formed per mg protein per min (mean value ± SEM; n = 5). Statistical significance, *p < 0.01.

Article Snippet: DHEA, 7a-OH-DHEA, 7b-OY-DHEA and 7-oxo-DHEA, 11a-hydroxy-progesterone (11b-OH-PRO), 11b-hydroxy-progesterone (11b-OH-PRO), CS and DHC were obtained from Steraloids Inc. (Newport, RI).

Techniques: Incubation, Inhibition

Journal: Archives of biochemistry and biophysics

Article Title: A novel NADP(+)-dependent dehydrogenase activity for 7alpha/beta- and 11beta-hydroxysteroids in human liver nuclei: A third 11beta-hydroxysteroid dehydrogenase.

doi: 10.1016/j.abb.2009.04.010

Figure Lengend Snippet: Fig. 5. Effects of inhibitors on the reductive metabolism of 7-oxo-DHEA to 7a-OH- DHEA and 7b-OH-DHEA by human liver microsomal protein in the presence of 1 mM b-NADPH. Human liver microsomal protein was incubated for 120 min with 100 lM 7-oxo-DHEA in the absence or presence of either CBX (2 lM) or 50 lM of either 11a-OH-PRO, 11b-OH-PRO, DHC, CS or 7a-OH-DHEA as described in the method section. The reaction rates was expressed as % control the rates of reduction of 7oxo-DHEA (mean value ± SEM; n = 5). Statistical significance, *p < 0.01.

Article Snippet: DHEA, 7a-OH-DHEA, 7b-OY-DHEA and 7-oxo-DHEA, 11a-hydroxy-progesterone (11b-OH-PRO), 11b-hydroxy-progesterone (11b-OH-PRO), CS and DHC were obtained from Steraloids Inc. (Newport, RI).

Techniques: Incubation, Control

Journal: Archives of biochemistry and biophysics

Article Title: A novel NADP(+)-dependent dehydrogenase activity for 7alpha/beta- and 11beta-hydroxysteroids in human liver nuclei: A third 11beta-hydroxysteroid dehydrogenase.

doi: 10.1016/j.abb.2009.04.010

Figure Lengend Snippet: Fig. 6. The effects of inhibitors on the conversion of CS to DHC or 7a-OH-DHEA to 7- oxo-DHEA by human liver microsomal and nuclear protein with 1 mM b-NADP+. Human liver microsomal or nuclear fraction protein (0.5 mg/mL) was incubated with 1 mM b-NADP and either 80 nM CS or 100 lM 7a-OH-DHEA for 120 min in the absence or presence of CBX (2 lM) or 50 lM of either 11a-OH-PRO, 11b-OH-PRO, DHC, CS or 7-oxo-DHEA as described in the method section. (A, B) Human liver nuclear protein (ND, not determined), (C) Human liver microsomal protein. The rates of sterol oxidation were expressed as nmol DHC formed per mg per min or pmol 7-oxo-DHEA formed per mg protein per minute (mean value ± SEM; n = 5). Statistical significance, *p < 0.01.

Article Snippet: DHEA, 7a-OH-DHEA, 7b-OY-DHEA and 7-oxo-DHEA, 11a-hydroxy-progesterone (11b-OH-PRO), 11b-hydroxy-progesterone (11b-OH-PRO), CS and DHC were obtained from Steraloids Inc. (Newport, RI).

Techniques: Incubation