Multi-omics and experimental validation reveal the mechanism of DanxiaTiaoban decoction in treating atherosclerosis.

Exploring the therapeutic potential of Xiangsha Liujunzi Wan in Crohn's disease: from network pharmacology approach to experimental validation

BackgroundTriple-negative breast cancer is a biological subtype of breast cancer, which is unresponsive to conventional chemotherapies and has a poor prognosis. C-Phycocyanin (C-PC), a marine natural purified from Spirulina platensis, has been investigated that has anti-cancer function. The mitogen activated protein kinase (MAPK) pathway plays a crucial role in the development and progression of cancer. Therefore, we would like to study the anti-cancer effects of C-phycocyanin in the treatment of triple-negative breast cancer, and explore the role of MAPK pathway in the anti-tumor effects of C-phycocyanin.MethodsCell proliferation, cell cycle, cell apoptosis and cell migration were explored in breast cancer MDA-MB-231 cell lines. AKT, MAPK and membrane death receptor signaling were evaluated in MDA-MB-231 cell lines.ResultsOur study indicated that C-phycocyanin inhibited cell proliferation and reduced colony formation ability of MDA-MB-231 cells. Furthermore, C-phycocyanin induced cell cycle G0/G1 arrest by decreasing protein expression levels of Cyclin D1 and CDK-2 and increasing protein expression levels of p21 and p27. In addition, C-phycocyanin induced cell apoptotic by activating cell membrane surface death receptor pathway. Besides, C-phycocyanin down-regulated the protein expression levels of cyclooxygenase-2, and further inhibited MDA-MB-231 cells migration. We also found cell death induced by C-phycocyanin was carried through the MAPK signaling pathways. C-Phycocyanin was able to induce MDA-MB-231 cell apoptosis by activating p38 MAPK and JNK signaling pathways while inhibiting ERK pathway.ConclusionsC-Phycocyanin exerted anti-cancer activity via the MAPK signaling pathway in MDA-MB-231 cells.

To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) signaling pathway in the expression of aquaporin 4 (AQP4) in brain tissue of rats with cardiopulmonary resuscitation (CPR) during mild hypothermia. Forty-eight healthy male Sprague-Dawley (SD) rats were divided into sham operation group, normal temperature group and mild hypothermia group according to random number table method, with 16 in each group. The rat model of cardiac arrest-cardiopulmonary resuscitation (CPR) was established by asphyxia method. The sham operation group only experienced venous catheterization and tracheal intubation. The mild hypothermia group was treated with hypothermia 0.5 hours after restore of spontaneous circulation (ROSC, maintaining esophageal temperature at 32-34 centigrade); the normal temperature group was treated at room temperature after ROSC (maintaining esophageal temperature at 36-38 centigrade). Brain tissue was harvested at 6 hours after ROSC, and histopathological changes were observed by hematoxylin-eosin (HE) staining. The water content of brain tissue was determined by dry-wet specific gravity method. The protein expressions of phosphorylation of p38 mitogen-activated protein kinase (p-p38MAPK), p38MAPK and AQP4 in brain tissue were determined by Western Blot. Compared with the sham operation group, the nerve cells in the normal temperature group were reduced in size, cytoplasmic loosening, nuclear pyknosis, and in apoptotic body formation, water content of brain tissue was significantly increased [(83.64±2.53)% vs. (77.95±0.94)%, P < 0.05], the protein expressions of p-p38MAPK, p38MAPK, AQP4 were significantly increased (p38MAPK/β-actin: 1.010±0.217 vs. 0.427±0.090, p-p38MAPK/p38MAPK: 0.451±0.172 vs. 0.191±0.141, AQP4/β-actin: 3.129±0.754 vs. 1.598±0.464, all P < 0.05). Compared with the normal temperature group, the degree of necrosis of nerve cells in the mild hypothermia group was reduced, the water content of brain tissue was significantly decreased [(80.49±2.05)% vs. (83.64±2.53)%, P < 0.05], the protein expression of p38MAPK, p-p38MAPK and AQP4 in brain tissue were significantly decreased (p38MAPK/β-actin: 0.590±0.162 vs. 1.010±0.217, p-p38MAPK/p38MAPK: 0.298±0.076 vs. 0.451±0.172, AQP4/β-actin: 2.061±0.340 vs. 3.129±0.754, all P < 0.05). Mild hypothermia may regulate the expression of AQP4 in brain tissue of CPR rats through p38MAPK signaling pathway, and reduce brain edema, thereby exerting brain protection.

Nisin Z is a possible alternative for treating bovine mastitis by inhibiting mastitis-causing pathogens and having anti-inflammatory activity. However, the anti-inflammatory mechanism of nisin Z on mastitis is unknown. Our study aimed to investigate the mechanisms of nisin Z on mastitis. Our results showed that nisin Z inhibited the activation of the ERK1/2 and p38 mitogen-activated protein kinase (MAPK) signaling pathway, decreased the release of pro-inflammatory cytokines (i.e., tumor necrosis factor-α, IL-1β, and IL-6), and increased the anti-inflammatory cytokine (IL-10) in lipopolysaccharide (LPS)-induced MCF10A cells. After intraperitoneal injection, nisin Z significantly decreased inflammatory cell infiltration in the mammary gland, as well as decreased myeloperoxidase and pro-inflammatory cytokines in serum and mammary gland. Western blot analysis revealed that nisin Z also dramatically suppressed the activation of the ERK1/2 and p38 MAPK signaling pathways in LPS-induced mastitis mice. We also found that nisin Z treatment could enhance the blood-milk barrier. In summary, our study demonstrated that nisin Z exerted an anti-inflammatory effect by inhibiting the ERK1/2 and p38 MAPK signaling pathway and promoting the blood-milk barrier on LPS-induced mastitis.

Lung cancer is the main health threat in the world. Recently, oleuropein has been reported to have potent antioxidant and anticancer activities. However, the antitumor effects of oleuropein on H1299 cells are not well understood. Therefore, the purpose of this paper is tantamount to explore the effects of oleuropein on H1299 cells and its underlying mechanism that may be involved. Oleuropein treatment in H1299 cells resulted in cell cycle distribution at G2 /M arrest and apoptosis in a dose-dependent manner. Mitochondria-mediated apoptosis was verified by the increase in Bax/Bcl-2 ratio, release of cytochrome c, and activation of caspase-3 on oleuropein-induced H1299 cells. In addition, our data also demonstrated that the p38 mitogen-activated protein kinase (MAPK) signaling pathway has a critical role in oleuropein-induced apoptosis. Moreover, we used transcriptome analysis to identify differentially expressed genes (DEGs) in H1299 cells by oleuropein and SB203580 treatment. Many DEGs were annotated to metabolic pathways, cell cycle, pathways in cancer, MAPK signaling pathway by Kyoto Encyclopedia of Genes and Genomes and Gene ontology enrichment methods. Network and expression analysis found that DEGs, including RPS6A5, GADD45A, and MKP, play a key role in the p38 MAPK signaling pathway. In H1299 cells, oleuropein resulted in the expression of numerous genes related to cell signaling, metabolism pathway and directly associated with apoptosis. These results illustrated that oleuropein-induced apoptosis via mitochondrial apoptotic cascade was activated by the p38 MAPK signaling pathway in H1299 cells. Thus, oleuropein as a natural compound and therapeutic drug has potential application value in the treatment of lung cancer.

BackgroundHuangqi Guizhi Wuwu decoction (HQGZWWD) has been used to treat and prevent deep vein thrombosis (DVT) in China. However, its potential mechanisms of action remain unclear. This study aimed to utilize network pharmacology and molecular docking technology to elucidate the molecular mechanisms of action of HQGZWWD in DVT.MethodsWe identified the main chemical components of HQGZWWD by reviewing the literature and using a Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. We used GeneCards and Online Mendelian Inheritance in Man databases to identify the targets of DVT. Herb-disease-gene-target networks using Cytascape 3.8.2 software; a protein–protein interaction (PPI) network was constructed by combining drug and disease targets on the STRING platform. Additionally, we conducted Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Finally, molecular docking verification of active components and core protein targets was conducted.ResultsA total of 64 potential targets related to DVT were identified in HQGZWWD, with 41 active components; quercetin, kaempferol, and beta-sitosterol were the most effective compounds. The PPI network analysis revealed that AKT1, IL1B, and IL6 were the most abundant proteins with the highest degree. GO analysis indicated that DVT treatment with HQGZWWD could involve the response to inorganic substances, positive regulation of phosphorylation, plasma membrane protein complexes, and signaling receptor regulator activity. KEGG analysis revealed that the signaling pathways included pathways in cancer, lipid and atherosclerosis, fluid shear stress and atherosclerosis, and the phosphatidylinositol 3-kinases/protein kinase B(PI3K-Akt) and mitogen-activated protein kinase (MAPK) signaling pathways. The molecular docking results indicated that quercetin, kaempferol, and beta-sitosterol exhibited strong binding affinities for AKT1, IL1B, and IL6.ConclusionOur study suggests that AKT1, IL1B, and IL6 are promising targets for treating DVT with HQGZWWD. The active components of HQGZWWD likely responsible for its effectiveness against DVT are quercetin, kaempferol, and beta-sitosterol, they may inhibit platelet activation and endothelial cell apoptosis by regulating the PI3K/Akt and MAPK signaling pathways, slowing the progression of DVT.

Electroacupuncture attenuates cerebral hypoxia and neuronal apoptosis induced by cerebral ischemia/reperfusion injury. To further identify the involved mechanisms, we assumed that electroacupuncture used to treat cerebral ischemia/reperfusion injury was associated with the p38 mitogen-activated protein kinase (MAPK) signaling pathway. We established rat models of cerebral ischemia/reperfusion injury using the modified Zea-Longa's method. At 30 minutes before model establishment, p38 MAPK blocker SB20358 was injected into the left lateral ventricles. At 1.5 hours after model establishment, electroacupuncture was administered at acupoints of Chize (LU5), Hegu (LI4), Zusanli (ST36), and Sanyinjiao (SP6) for 20 minutes in the affected side. Results showed that the combination of EA and SB20358 injection significantly decreased neurologic impairment scores, but no significant differences were determined among different interventional groups. Hematoxylin-eosin staining also showed reduced brain tissue injuries. Compared with the SB20358 group, the cells were regularly arranged, the structures were complete, and the number of viable neurons was higher in the SB20358 + electroacupuncture group. Terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling assay showed a decreased apoptotic index in each group, with a significant decrease in the SB20358 + electroacupuncture group. Immunohistochemistry revealed reduced phosphorylated p38 expression at 3 days in the electroacupuncture group and SB20358 + electroacupuncture group compared with the ischemia/reperfusion group. There was no significant difference in phosphorylated p38 expression between the ischemia/reperfusion group and SB20358 group. These findings confirmed that the electroacupuncture effects on mitigating cerebral ischemia/reperfusion injury are possibly associated with the p38 MAPK signaling pathway. A time period of 3 days could promote the repair of ischemic cerebral nerves.

The Effects of Mitogen-activated Protein Kinase Signaling Pathways on Lipopolysaccharide-mediated Osteo/Odontogenic Differentiation of Stem Cells from the Apical Papilla

Objective To explore the regulatory effects of ultraviolet B (UVB) radiation on the mitogenactivated protein kinase (MAPK) signaling pathway in cultured HaCaT cells in vitro at different post-radiation time points. Methods Some cultured HaCaT cells were classified into several groups to be exposed to UVB of 1.5, 4.5, 7.5, 10, 20, 30 and 50 mJ/cm2 for 1, 3, 5, 7, 15, 20 and 34 seconds respectively, or UVB of 50 mJ/cm2 for 34 seconds, or remain untreated (control group). Western blot was performed to determine the total and phosphorylation levels of ERK1/2, JNK and p38 in HaCaT cells at 8 hours after exposure to 1.5, 4.5, 7.5,10, 20, 30 and 50 mJ/cm2 UVB, as well as at 2, 4, 8 and 12 hours after exposure to 50 mJ/cm2 UVB. Quantity One software was used to measure the absorbance value of protein bands, and protein levels were expressed as the absorbance ratio of target bands to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Statistical analysis was carried out by using one- way analysis of variance, least significant difference (LSD) - t test and multivariate analysis of variance. Results Compared with the control group, there was a significant increase at 8 hours in the phosphorylation levels of ERK 1/2 and JNK in HaCaT cells after exposure to 7.5, 10, 20, 30 and 50 mJ/cm2 UVB (all P< 0.05), and in the phosphorylation level of p38 after exposure to 20, 30 and 50 mJ/cm2 UVB (all P < 0.05), with the strongest increase in phosphorylation levels of ERK 1/2, JNK and p38 all observed in HaCaT cells exposed to 50 mJ/cm2 UVB (all P< 0.05). After exposure to 50 mJ/cm2 UVB, the phosphorylation level of ERK1/2 began to increase in HaCaT cells at 8 hours, and significantly increased at 12 hours(44.844±2.023 vs. 10.277±0.320, P< 0.05) compared with the control group; however, the phosphorylation levels of p38 and JNK increased in HaCaT cells as early as 2 hours after exposure (both P< 0.05), and remained significantly higher compared with the control group at 4, 8 and 12 hours (all P< 0.05), while the increasing trend in phosphorylated JNK level weakened over time (P< 0.05). Conclusions The ERK1/2, JNK and p38 MAPK signaling pathways were activated in HaCaT cells after exposure to 50 mJ/cm2 UVB, and the degree of their activation varied over time. Key words: Ultraviolet rays; Cells, cultured; Keratinocytes; Signal transduction; MAP kinase kinase kinases

Antiglycative activity of sulforaphane: a new avenue to counteract neurodegeneration?

Airway remodeling is the main characteristic of asthma; however, the mechanisms underlying this pathophysiological change have not been fully elucidated. Previous studies have indicated that the Wnt/β-catenin and mitogen-activated protein kinase (MAPK) signaling pathway are involved in the development of airway remodeling during asthma. Therefore, the present study established an airway remodeling rat model, after which β-catenin, cyclin D1 and c-Myc protein expressions were analyzed via western blotting in the lung tissue and airway smooth muscle cells (ASMCs) of rats. The mRNA expression of the aforementioned proteins were evaluated via reverse transcription-quantitative PCR. β-catenin, cyclin D1 and c-Myc are core transcription factors and target genes of the Wnt/β-catenin and MAPK signaling pathways. Furthermore, β-catenin, c-Myc and cyclin D1 protein expression were determined following blocking of the p38 MAPK signaling pathway in vitro. The results demonstrated that higher expressions of β-catenin, cyclin D1 and c-Myc were detected in lung tissues and ASMCs in the asthma group compared with the control. Blocking the p38 MAPK signaling pathway with a specific inhibitor SB203580 also downregulated the expressions of β-catenin, cyclin D1 and c-Myc in vitro. Taken together, these results indicated that the Wnt/β-catenin signaling pathway may regulate the process of airway remodeling via the p38 MAPK-dependent pathway.

This paper investigated the effect of total saponins from Rhizoma Panacis Majoris on the proliferation, apoptosis, and autophagy of human cervical carcinoma HeLa cells. The saponin content was detected by ultraviolet-visible spectrophotometry. Cell coun-ting kit-8(CCK-8) assay, 4,6-diamidino-2-phenylindole(DAPI) staining, and flow cytometry were used to detect the effects of total saponins of Panacis Majoris Rhizoma on cell viability, morphology, cell cycle and apoptosis of HeLa cells. Western blot was used to detect the expression of apoptosis-related proteins B cell lymphoma-2(Bcl-2), Bcl-2-associated X protein(Bax), cleaved caspase-9, and cleaved caspase-3, autophagy-related proteins Beclin-1 and SQSTM1(p62), and the proteins related to the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR) and mitogen-activated protein kinase(MAPK) signaling pathways. It was found that the yield and saponin content of total saponins from Rhizoma Panacis Majoris were 6.3% and 78.3%, respectively. Total saponins from Rhizoma Panacis Majoris could significantly inhibit the proliferation(P&lt;0.001), effect the nuclear morphology, block the G_0/G_1 cycle, and induce cell apoptosis in HeLa cells with a concentration-dependent manner. In addition, total saponins from Rhizoma Panacis Majoris up-regulated the expression of pro-apoptotic proteins Bax, cleaved caspase-9, and cleaved caspase-3, and autophagy-related protein p62(P&lt;0.05), while down-regulated the expression of anti-apoptotic protein Bcl-2 and autophagy-related protein Beclin-1(P&lt;0.01). Total saponins from Rhizoma Panacis Majoris could promote the expression of p-p38/p38, p-Jun N-terminal kinase(JNK)/JNK, p-PI3K/PI3K, p-Akt/Akt, p-mTOR/mTOR proteins in PI3K/Akt/mTOR and MAPK signaling pathways(P&lt;0.05). In contrast, the effect on p-ERK/ERK expression was not obvious. Therefore, total saponins from Rhizoma Panacis Majoris may inhibit autophagy and promote apoptosis of HeLa cells through the activation of the PI3K/Akt/mTOR, c-JNK, and p38 MAPK signaling pathways, which indicates that total saponins from Rhizoma Panacis Majoris may have a potential role in cervical cancer treatment.

Hypertensive disorders complicating pregnancy (HDCP) are one of the most serious medical disorders during pregnancy. To investigate the effects of hydrogen on the mitogen-activated protein kinase (MAPK) signaling pathway in preeclampsia (PE). The N(omega)-nitro-L-arginine methyl ester (L-NAME)-induced PE model with Sprague Dawley (SD) rats was employed. An inhibitor of MAPK signaling pathways (SB203580) was used as a p38 MAPK inhibitor. The SD rats were randomized into 5 groups: non-pregnant (NP); normal pregnancy (P); pregnancy + L-NAME (L); pregnancy + L-NAME + hydrogen-rich saline (LH); and pregnancy + L-NAME + hydrogen-rich saline + SB203580 (LHS). The pregnancies were terminated on day 22 of gestation, and the placentas and kidneys were microscopically inspected. Tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β) and malondialdehyde (MDA) levels were assessed. The mean systolic blood pressure (SBP) and level of proteinuria were recorded. The p38 MAPK mRNA expression and p-p38 MAPK protein levels were measured using real-time polymerase chain reaction (RT-PCR) and western blot, respectively. It was found that hydrogen-rich saline (LH group) decreased placental MDA, proteinuria, TNF-α, and IL-1β levels in the placental tissues compared with the L group (all p < 0.05). Additionally, hydrogen-rich saline (LH group) treatment significantly decreased the p38 MAPK mRNA expression and p-p38 MAPK protein levels compared with the L group (p < 0.05). The p38 MAPK inhibitor SB203580 (LHS group) further decreased the p38 MAPK mRNA expression and p-p38 MAPK protein levels compared with the LH group (p < 0.05). Hydrogen can decrease the reactive oxygen species (ROS) content and inhibit the MAPK pathway. The protective effect of hydrogen may be associated with the inhibition of the p38 MAPK signaling pathway.

Causal relationship between circulating inflammatory proteins and atherosclerosis: a bidirectional Mendelian randomization study and meta-analysis.

BackgroundPrior studies have suggested a significant connection between fasting insulin (FI) and androgenetic alopecia (AGA), but the exact cause of this connection and underlying molecular mechanism has not been clarified. In this study, a Mendelian randomization (MR) analysis was utilized to discover the causal associations between FI and AGA.MethodsGenome-wide association study (GWAS) data for FI and AGA were retrieved, and bidirectional MR analysis was conducted. FI-associated genes were identified through expression quantitative trait loci (eQTL) analysis, with enrichment analysis and a protein-protein interaction (PPI) network used to explore potential pathways and core genes.ResultsForward MR analysis revealed a significant causal relationship between elevated FI levels and AGA (P=0.027, OR=43.944). Reverse MR analysis found no causal effect of AGA on FI (P=0.808, OR=1.0001). A total of 92 FI-associated genes were analyzed, with enrichment results indicating involvement in glycine, serine, and threonine metabolic pathways. EIF2B4 and NRBP1 were identified as potential core genes linking FI and AGA.ConclusionBy using MR analysis, this study verified the possible causative connection between FIns and AGA by MR analysis. The core genes EIF2B4 and NRBP1, along with biological processes such as glycosylation and amino acid metabolism, may serve as crucial links.