Journal: Frontiers in Pharmacology

Article Title: Salidroside Improves Chronic Stress Induced Depressive Symptoms Through Microglial Activation Suppression

doi: 10.3389/fphar.2021.635762

Figure Lengend Snippet: SLDS suppressed microglia activation and pro-inflammatory cytokines release in hippocampus of chronic stress exposed mice. (A) Representative images illustrating microglia stained with Iba-1 and CD68 antibodies in hippocampal sections. Scale bar, 100 µm; (B) Number of Iba-1 positive microglia in hippocampal sections [F(2,27) = 1.515, P = 0.2568]; (C) Number of CD68 positive microglia in hippocampal sections [F(2,27) = 151.9, P < 0.0001]; (D) mRNA level of IL-1β [F(2,9) = 35.23, P < 0.0001] , IL-6 [F(2,9) = 19.28, P = 0.0006] , IL-18 [F(2,9) = 9.481, P = 0.0061] and TNF-α [F(2,9) = 12.61, P = 0.0025] in hippocampus determined by qRT-PCR ( n = 4); (E) The secretion level of IL-1β [F(2,15) = 8.063, P = 0.0042], IL-6 [F(2,15) = 29.19, P < 0.0001], IL-18 [F(2,15) = 5.890, P = 0.0129] and TNF-α [F(2,15) = 24.66, P < 0.0001] in hippocampus determined using ELISA ( n = 4); (F) P-ERK1/2 [F(2,9) = 10.15, P = 0.0049] in hippocampal area tissue; (G) P-p38 MAPK [F(2,12) = 13.05, P = 0.0010] in hippocampal area tissue; (H) P-p65 NF-κB [F(2,15) = 28.59, P < 0.0001] in hippocampal area tissue; (I) iNOS expression [F(2,15) = 14.42, P = 0.0003] in hippocampal area tissue. All data are presented as mean ± SD. * P < 0.05, ** P <0.01, *** P <0.005, compare to control group; # P < 0.05, ## P <0.01, ### P < 0.005, compare to stress group.

Article Snippet: Rabbit polyclonal antibodies against phospho-p42/44 MAPK, p42/44 MAPK, phospho-p38 MAPK, p38 MAPK, phospho-NF-κB p65, NF-κB p65, iNOS (1:5000, CST), phospho-paxillin Ser83 and mouse polyclonal antibodies against paxillin (1:2500, ECM biosciences), α-Tubulin (1:10000, CST) were used.

Techniques: Activation Assay, Staining, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Salidroside Improves Chronic Stress Induced Depressive Symptoms Through Microglial Activation Suppression

doi: 10.3389/fphar.2021.635762

Figure Lengend Snippet: SLDS reduced the levels of proinflammatory cytokines and suppressed microglial activation in LPS induced primary microglia. (A) The cell viability with SLDS (0, 50, 100 and 200 μM) alone [F(3,12) = 1.524, P = 0.2588] in primary microglia; (B) The cell viability with SLDS (0, 50, 100 and 200 μM) and LPS 1 μg/ml [F(4,31) = 0.6963, P = 0.6003] in primary microglia; (C) mRNA levels of IL-1β [F(2,9) = 17.54, P = 0.0008] , I L-6 [F(2,9) = 25.80, P = 0.0002] , IL-18 [F(2,9) = 8.516, P = 0.0084] and TNF-α [F(2,9) = 81.80, P < 0.0001] in primary microglia ( n = 4); (D) The secretion levels of IL-1β [F(2,21) = 57.51, P = < 0.0001], IL-6 [F(2,21) = 19.17, P < 0.0001], IL-18 [F(2,21) = 21.64, P < 0.0001] and TNF-α [F(2,21) = 23.71, P < 0.0001] in primary microglia ( n = 4); (E) P-ERK1/2 [F(2,15) = 72.91, P < 0.0001] in primary microglia; (F) P-p38 MAPK [F(2,12) = 17.38, P = 0.0003] in primary microglia; (G) P-p65 NF-κB [F(2,9) = 86.64, P < 0.0001] in primary microglia; (H) iNOS expression [F(2,11) = 12.46, P = 0.0015] in primary microglia. All data are presented as mean ± SD. ** P <0.01, *** P <0.005, compare to control group; # P < 0.05 ## P <0.01 ### P < 0.005, compare to LPS treated group.

Article Snippet: Rabbit polyclonal antibodies against phospho-p42/44 MAPK, p42/44 MAPK, phospho-p38 MAPK, p38 MAPK, phospho-NF-κB p65, NF-κB p65, iNOS (1:5000, CST), phospho-paxillin Ser83 and mouse polyclonal antibodies against paxillin (1:2500, ECM biosciences), α-Tubulin (1:10000, CST) were used.

Techniques: Activation Assay, Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Salidroside Improves Chronic Stress Induced Depressive Symptoms Through Microglial Activation Suppression

doi: 10.3389/fphar.2021.635762

Figure Lengend Snippet: Anti-inflammation mechanism of SLDS in LPS induced primary microglia. LPS signal from infections activates microglia through TLRs and further activates ERK1/2, p38 MAPK, NF-κB signaling. Signals translocate to nucleus and induce the transcription of inflammatory cytokines IL-1β, IL-6, IL-18 and TNF-α. On the other hand, cytoskeleton changes generate the mechanical forces that drive cell motility and phagocytosis which are closely associated with microglial function. LPS signals induce the cytoskeleton protein vimentin filaments assembly which leads to the morphological changes from ramified state to ameboid state. Meanwhile, they regulate focal adhesion assembly which results in the cell attachment state alteration. SLDS can inhibit the LPS signals transduction thus blocking the activation of microglia and prevent cytoskeleton changes.

Article Snippet: Rabbit polyclonal antibodies against phospho-p42/44 MAPK, p42/44 MAPK, phospho-p38 MAPK, p38 MAPK, phospho-NF-κB p65, NF-κB p65, iNOS (1:5000, CST), phospho-paxillin Ser83 and mouse polyclonal antibodies against paxillin (1:2500, ECM biosciences), α-Tubulin (1:10000, CST) were used.

Techniques: Cell Attachment Assay, Transduction, Blocking Assay, Activation Assay