Journal: International Journal of Molecular Sciences

Article Title: Implication of lncRNA MSTRG.81401 in Hippocampal Pyroptosis Induced by P2X7 Receptor in Type 2 Diabetic Rats with Neuropathic Pain Combined with Depression

doi: 10.3390/ijms25021186

Figure Lengend Snippet: Expression of inflammasome/pyroptosis pathway proteins ( A – G ) and inflammatory factors ( H – M ) in hippocampus was examined by Western blotting. The bar graphs show the statistical data of relative expression levels of targeted molecules after being normalized by β-actin. NLRP3, GSDMD and ASC ( A – D ); procaspase-1 and cleaved-caspase-1 ( E – G ); TNF-α ( H , I ), IL-1β ( J , K ) and IL-18 ( L , M ). Values are mean ± SEM, n = 6; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Ctrl group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. DNP + MDD group.

Article Snippet: The primary antibodies used and their sources are as follows: P2X7 (1:800, APR-008, Alomone, Jerusalem, Israel), NLRP3 (1:800, IMG-6668A, Novus Biologicals, Centennial, CO, USA), GSDMD (1:800, NBP2-33422, Novus Biologicals, CO, USA), ASC (1:500, DF6304, Affinity Biosciences, Cincinnati, OH, USA), pro-caspase-1 (1:1000, AB179515, Abcam, Cambridge, UK), cleaved-caspase-1 (1:1000, AF4022, Affinity Biosciences, OH, USA), IL-1β (1:500, BA14789, Boster, Pleasanton, CA, USA), TNF-α (1:500, BA0131, Boster, CA, USA), IL-18 (1:500, DF6252, Affinity Biosciences, OH, USA), and β-actin (1:1000, Zhong Shan-Gold Bridge, Beijing, China).

Techniques: Expressing, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Implication of lncRNA MSTRG.81401 in Hippocampal Pyroptosis Induced by P2X7 Receptor in Type 2 Diabetic Rats with Neuropathic Pain Combined with Depression

doi: 10.3390/ijms25021186

Figure Lengend Snippet: The serum concentrations of TNF-α ( A ), IL-18 ( B ) and IL-1β ( C ) were assessed using ELISA. Values are mean ± SEM, n = 6; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Ctrl group; ## p < 0.01, ### p < 0.001 vs. DNP + MDD group.

Article Snippet: The primary antibodies used and their sources are as follows: P2X7 (1:800, APR-008, Alomone, Jerusalem, Israel), NLRP3 (1:800, IMG-6668A, Novus Biologicals, Centennial, CO, USA), GSDMD (1:800, NBP2-33422, Novus Biologicals, CO, USA), ASC (1:500, DF6304, Affinity Biosciences, Cincinnati, OH, USA), pro-caspase-1 (1:1000, AB179515, Abcam, Cambridge, UK), cleaved-caspase-1 (1:1000, AF4022, Affinity Biosciences, OH, USA), IL-1β (1:500, BA14789, Boster, Pleasanton, CA, USA), TNF-α (1:500, BA0131, Boster, CA, USA), IL-18 (1:500, DF6252, Affinity Biosciences, OH, USA), and β-actin (1:1000, Zhong Shan-Gold Bridge, Beijing, China).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Therapeutic effects of 1,25-dihydroxyvitamin D3 on diabetes-induced liver complications in a rat model

doi: 10.3892/etm.2016.3242

Figure Lengend Snippet: Primers sequences for reverse transcription-quantitative polymerase chain reaction analysis.

Article Snippet: Tissue slices were incubated with rabbit anti-rat antibodies against C-Jun (bs-0670R; 1:100; Beijing Biosynthesis Biotechnology, Co., Ltd., Beijing, China), JNK, TNF-α and IL-1β (BA1648, BA0131 and BA2782; 1:100; Wuhan Boster Biological Technology, Ltd., Wuhan, China) respectively at 4°C overnight.

Techniques: Polymerase Chain Reaction, Sequencing

Journal: Experimental and Therapeutic Medicine

Article Title: Therapeutic effects of 1,25-dihydroxyvitamin D3 on diabetes-induced liver complications in a rat model

doi: 10.3892/etm.2016.3242

Figure Lengend Snippet: Changes in liver histology (magnification, ×400; stain, hematoxylin and eosin). In the NC group, liver cell alignment is normal. In the DM group, liver cells appear swelled with fatty degeneration and inflammatory cell infiltration. In the VD group, fatty degeneration and inflammatory lesions are alleviated, in comparison with the DM group. NC, normal control group; DM, Type 2 diabetes mellitus group; VD, vitamin D-treated group.

Article Snippet: Tissue slices were incubated with rabbit anti-rat antibodies against C-Jun (bs-0670R; 1:100; Beijing Biosynthesis Biotechnology, Co., Ltd., Beijing, China), JNK, TNF-α and IL-1β (BA1648, BA0131 and BA2782; 1:100; Wuhan Boster Biological Technology, Ltd., Wuhan, China) respectively at 4°C overnight.

Techniques: Staining, Comparison, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Therapeutic effects of 1,25-dihydroxyvitamin D3 on diabetes-induced liver complications in a rat model

doi: 10.3892/etm.2016.3242

Figure Lengend Snippet: Immunohistochemical analysis of the protein expression of target genes in the liver tissues of type 2 diabetes mellitus model rats (magnification, ×400; stain, hematoxylin and eosin). JNK, TNF-α and IL-1β are predominantly expressed in the cytoplasm, while C-Jun is primarily expressed in the nucleus. For JNK, C-Jun, TNF-α and IL-1β there is obvious brown staining; protein expression levels are increased in the DM group compared with the NC group. Conversely, compared with the DM group, VD administration decreased the positive areas and staining intensity of each target protein. JNK, c-Jun N-terminal kinase; TNF-α, tumor necrosis factor-α; IL-1β, interleukin-1β; NC, normal control group; DM, type 2 diabetes mellitus group; VD, vitamin D-treated group.

Article Snippet: Tissue slices were incubated with rabbit anti-rat antibodies against C-Jun (bs-0670R; 1:100; Beijing Biosynthesis Biotechnology, Co., Ltd., Beijing, China), JNK, TNF-α and IL-1β (BA1648, BA0131 and BA2782; 1:100; Wuhan Boster Biological Technology, Ltd., Wuhan, China) respectively at 4°C overnight.

Techniques: Immunohistochemical staining, Expressing, Staining, Control

Journal: Experimental and Therapeutic Medicine

Article Title: Therapeutic effects of 1,25-dihydroxyvitamin D3 on diabetes-induced liver complications in a rat model

doi: 10.3892/etm.2016.3242

Figure Lengend Snippet: Immunohistochemical staining for TNF-α and IL-1β protein expression levels in the rat liver tissues.

Article Snippet: Tissue slices were incubated with rabbit anti-rat antibodies against C-Jun (bs-0670R; 1:100; Beijing Biosynthesis Biotechnology, Co., Ltd., Beijing, China), JNK, TNF-α and IL-1β (BA1648, BA0131 and BA2782; 1:100; Wuhan Boster Biological Technology, Ltd., Wuhan, China) respectively at 4°C overnight.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Therapeutic effects of 1,25-dihydroxyvitamin D3 on diabetes-induced liver complications in a rat model

doi: 10.3892/etm.2016.3242

Figure Lengend Snippet: Reverse transcription-quantitative polymerase chain reaction analysis of mRNA expression levels.

Article Snippet: Tissue slices were incubated with rabbit anti-rat antibodies against C-Jun (bs-0670R; 1:100; Beijing Biosynthesis Biotechnology, Co., Ltd., Beijing, China), JNK, TNF-α and IL-1β (BA1648, BA0131 and BA2782; 1:100; Wuhan Boster Biological Technology, Ltd., Wuhan, China) respectively at 4°C overnight.

Techniques: Polymerase Chain Reaction, Expressing

Journal: Frontiers in Pharmacology

Article Title: Maternal 25-Hydroxyvitamin D Deficiency Promoted Metabolic Syndrome and Downregulated Nrf2/CBR1 Pathway in Offspring

doi: 10.3389/fphar.2020.00097

Figure Lengend Snippet: Maternal 25-hydroxyvitamin D deficiency altered expression levels of Nrf2, CBR1, and inflammatory cytokine in offspring at week 0 and 16. (A) Nrf2 and CBR1 immunohistochemical staining in offspring liver and pancreas tissues obtained in normal group (Con) and 25-hydroxyvitamin D deficiency group (VDD) at week 0 and 16 (magnification, ×200). (B , C) Western blotting analysis was used to detect theNrf2 and CBR1 expression level at week 0 and 16 in offspring rat liver (B) and pancreas (C) . Densitometry data for Nrf2 and CBR1 from the blots shown were normalized for analysis to GAPDH, the loading control. (D, E) The Nrf2 and CBR1 levels in offspring liver (D) and pancreas (E) at week 0 and 16 were determined by qRT-PCR. (F) IL-1β, IL-6and TNF-α IH staining in offspring liver and pancreas tissues obtained in normal group (Con) and 25-hydroxyvitamin D deficiency group (VDD) at week 0 and 16 (magnification, ×200). Mean ± SD, n = 5 rats per group. *** P < 0.01, vs. control group. Normal group, Con, Con-1, and Con-2 represent different rats; maternal 25-hydroxyvitamin D deficiency group, VDD, VDD-1, and VDD-2 represent different rats; week, w.

Article Snippet: Nrf2 (ab186825, Abcam), CBR1 (ab89433, Abcam), IL-1β (ab9722, Abcam), IL-6 (21865-1-AP, Proteintech), and TNFα (BA0131, Boster) were used for the IHC detection of protein expression in the trimethylamine (TMA).

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot, Control, Quantitative RT-PCR