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1.
Article in English | WPRIM | ID: wpr-929054

ABSTRACT

Marine fungi are important members of the marine microbiome, which have been paid growing attention by scientists in recent years. The secondary metabolites of marine fungi have been reported to contain rich and diverse compounds with novel structures (Chen et al., 2019). Aspergillus terreus, the higher level marine fungus of the Aspergillus genus (family of Trichocomaceae, order of Eurotiales, class of Eurotiomycetes, phylum of Ascomycota), is widely distributed in both sea and land. In our previous study, the coral-derived A. terreus strain C23-3 exhibited potential in producing other biologically active (with antioxidant, acetylcholinesterase inhibition, and anti-inflammatory activity) compounds like arylbutyrolactones, territrems, and isoflavones, and high sensitivity to the chemical regulation of secondary metabolism (Yang et al., 2019, 2020; Nie et al., 2020; Ma et al., 2021). Moreover, we have isolated two different benzaldehydes, including a benzaldehyde with a novel structure, from A. terreus C23-3 which was derived from Pectinia paeonia of Xuwen, Zhanjiang City, Guangdong Province, China.


Subject(s)
Acetylcholinesterase/metabolism , Animals , Anthozoa/microbiology , Anti-Inflammatory Agents/pharmacology , Aspergillus/chemistry , Benzaldehydes/pharmacology , Mice , Signal Transduction
2.
Article in English | WPRIM | ID: wpr-928947

ABSTRACT

OBJECTIVE@#To investigate whether the antihypertensive mechanism of electroacupuncture (EA) is associated with attenuating phenotype transformation of vascular smooth muscle cells (VSMCs) via phosphoinositide3-kinase (PI3K)/protein kinase B (Akt) and mitogen-activated protein kinase (MAPK) signaling pathways.@*METHODS@#Eight Wistar-ktoyo (WKY) rats were set as normal blood pressure group (normal group). A total of 32 spontaneous hypertensive rats (SHRs) were randomly divided into 4 groups using random number tables: a model group, an EA group, an EA+PI3K antagonist group (EA+P group), and an EA+p38 MAPK agonist+extracellular signal-regulated kinase (ERK) agonist group (EA+M group) (n=8/group). SHRs in EA group, EA+P group and EA+M group received EA treatment 5 sessions per week for continuous 4 weeks, while rats in the normal and model groups were bundled in same condition. The systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP) of each rat was measured at 0 week and the 4th week. After 4-week intervention, thoracic aorta was collected for hematoxylin-eosin (HE) staining, immunohistochemistry [the contractile markers α-smooth muscle actin (α-SMA) and calponin and the synthetic marker osteopontin (OPN)] and Western blot [α-SMA, calponin, OPN, PI3K, phosphorylated-Akt (p-Akt), Akt, p-p42/44 ERK, total p42/44 ERK, p-p38 MAPK and total p38 MAPK].@*RESULTS@#EA significantly reduced SBP, DBP and MAP (P<0.01). HE staining showed that the wall thickness of thoracic aorta in EA group was significantly decreased (P<0.01). From results of immunohistochemistry and Western blot, EA increased the expression of α-SMA and calponin, and decreased the expression of OPN (P<0.01). In addition, the expression of PI3K and p-Akt increased (P<0.01), while the expression of p-p42/44 ERK and p-p38 MAPK decreased in EA group (P<0.01). However, these effects were reversed by PI3K antagonist, p38 MAPK agonist and ERK agonist.@*CONCLUSIONS@#EA was an effective treatment for BP management. The antihypertensive effect of EA may be related with inhibition of phenotypic transformation of VSMCs, in which the activation of PI3K/Akt and the repression of MAPK pathway were involved.


Subject(s)
Animals , Electroacupuncture , Extracellular Signal-Regulated MAP Kinases/metabolism , MAP Kinase Signaling System , Muscle, Smooth, Vascular , Phenotype , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Rats , Rats, Inbred SHR
3.
Article in Chinese | WPRIM | ID: wpr-930333

ABSTRACT

Objective:To investigate the effects of baicalein combined with 5-FU on the proliferation, apoptosis and migration of small cell lung cancer H466 cells, and further to explore its sensitization mechanism.Methods:H466 cells were cultured in vitro and divided into blank control group, baicalein single treatment group, 5-FU single treatment group and combined treatment group. CCK-8 experiment was used to detect cell survival rate in each group; Flow cytometry was used to detect cell apoptosis and changes in ROS levels; Western blot was used to detect changes in the protein expression of the MAPK pathway.Results:CCK-8 results showed that the survival rates of the four groups of cells after 24 h treatment were 114.3% ± 2.8%, 79.8% ± 3.4%, 57.6% ± 1.8%, and 40.3% ± 2.7%. Compared with the other three groups, the combined treatment group had stronger inhibitory effect on the proliferation of H446 cells, and the difference was statistically significant ( P<0.001) . The proportion of apoptotic H446 cells in the combined treatment group was 49.2%±3.8%, which was significantly different from 33.9%±4.3% in the 5-FU single treatment group, and the difference was statistically significant ( P<0.001) . The inhibition rate of H446 cell migration in the combined treatment group was higher than that in the 5-FU single treatment group, and the difference was statistically significant ( P <0.001) . Combined treatment of baicalein and 5-FU can up-regulate ROS levels in H446 cells, thereby regulating the MAPK signaling pathway. Conclusion:The combined use of baicalein and 5-FU can increase the level of ROS in lung cancer H446 cells, activate the JNK and p38 signaling pathways, inhibit the ERK signaling pathway, and enhance 5-FU's proliferation inhibition, apoptosis induction and migration inhibition effects on H446 cells, which provides a certain experimental basis for the application of baicalein in small cell lung cancer.

4.
Article in Chinese | WPRIM | ID: wpr-940479

ABSTRACT

ObjectiveTo explore the mechanism of Fuzi Lizhongwan alleviating the damage of chemotherapy-induced peripheral neuropathy (CIPN) mice caused by cisplatin based on mitogen-activated protein kinase (MAPK) signaling pathway. MethodA total of 40 female KM mice were randomized into blank group (distilled water, ig), model group (distilled water, ig), Fuzi Lizhongwan group (3.5 g·kg-1, ig), and aspirin group (0.026 g·kg-1, ig). Cisplatin (3 mg·kg-1, ip, 5 days) was used to induce CIPN in mice. Administration began while modeling and lasted 12 days. The general conditions and behaviors of mice were observed. After the last administration, samples were collected. Pathological changes of the soles were observed based on hematoxylin-eosin (HE) staining. Biochemical assay was employed to determine the levels of serum superoxide dismutase (SOD), hydrogen peroxide (H2O2), malondialdehyde (MDA), and nitric oxide (NO), enzyme-linked immunosorbent assay (ELISA) the content of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and glutathione peroxidase-3 (GPX-3) in kidney tissue, and Western blotting the expression of extracellular signal-regulated kinase1/2 (ERK1/2), phosphorylated-ERK1/2 (p-ERK1/2), p38 MAPK, and phosphorylated-p38 MAPK (p-p38 MAPK) in kidney tissue. ResultCompared with the blank group, model group demonstrated obvious pathological damage on the soles, hyperkeratosis of the epidermis with a basketweave pattern, atrophy of stratum spinosum, reduction of cells, and intracellular edema. Compared with the model group, Fuzi Lizhongwan significantly alleviated the pathological damage of the skin tissue of the soles. The model group showed lower body weight, mechanical pain threshold, thermal pain threshold (P<0.01), and SOD activity (P<0.05), higher content of H2O2, MDA, and NO (P<0.01), and higher expression of IL-6, IL-1β, and TNF-α (P<0.01) than the blank group. Fuzi Lizhongwan group demonstrated higher body weight, mechanical pain threshold, thermal pain threshold (P<0.01), and SOD activity (P<0.05), lower content of H2O2, MDA, and NO (P<0.05), and lower expression of IL-6, IL-1β, and TNF-α (P<0.01) than the model group. The expression of ERK1/2, p-ERK1/2, p38 MAPK, and p-p38 MAPK increased significantly (P<0.01) in the model group compared with that in the blank group, while the expression decreased significantly (P<0.01) in the Fuzi Lizhongwan group compared with that in the model group. ConclusionFuzi Lizhongwan can relieve the neurological injury of cisplatin-induced CIPN mice and increase the pain threshold of mice, possibly by regulating the MAPK signaling pathway and inhibiting inflammatory response and oxidative stress.

5.
Article in Chinese | WPRIM | ID: wpr-940457

ABSTRACT

ObjectiveTo observe the repair effect of Dahuanglingxian prescription (DHLX) on bile duct epithelial cells of rats. To explore whether its mechanism of action is to adjust the mutual binding of transforming growth factor -β (TGF-β) activated kinase 1(TAK1) and tumor necrosis factor receptor-associated factor 6 (TRAF6), and regulate the activation of the nuclear transcription factor -κB (NF-κB)/mitogen-activated protein kinase (MAPK) signaling pathway. MethodThe 20 SD rats were randomly divided into normal group and DHLX group, 10 rats in each group, were given saline and DHLX (320 mg·kg-1·d-1) for 8 days, to prepare normal serum and DHLX serum. Biliary epithelial cells were extracted from normal SD rats and divided into 9 groups: Normal group, model group (20 mg·L-1), LPS+DHLX group (20 mg·L-1+10% DHLX), LPS+PDTC group (20 mg·L-1+200 μmol·L-1), LPS+SB203580 group (20 mg·L-1+0.5 μmol·L-1), LPS+PDTC+SB203580 group (20 mg·L-1+200 μmol·L-1+0.5 μmol·L-1), LPS+PDTC+DHLX group (20 mg·L-1+200 μmol·L-1+10% DHLX serum), LPS+SB203580+DHLX group (20 mg·L-1+0.5 μmol·L-1+10% DHLX serum), LPS+PDTC+SB203580 +DHLX group (20 mg·L-1+200 μmol·L-1+0.5 μmol·L-1+10% DHLX serum). The microscopic observation of morphological changes in each group of cells after drug intervention. Enzyme-linked immunosorbent assay(ELISA) was used to detect the expression of (IL)-1β and IL-6 in each group of cells. Western blot detected the expression levels of TAK1 and TRAF6 proteins in each group of cells, Co-IP detected the interaction between TAK1 and TRAF6, and further observed the distribution and co-localization of TAK1 and TRAF6 using Laser confocal microscope. ResultAfter the action of LPS, the cell synapses are reduced, the cell body becomes significantly rounded and smaller, but the cell morphology of each group tends to be normal after medication. Compared with normal group, the expression levels of IL-1β and IL-6 in model group were significantly increased (P<0.05), while the expression level of TAK1 was decreased while the expression level of TRAF6 was increased (P<0.05). The content of TAK1-TRAF6 protein complex showed a decreasing trend, and the two proteins co-located in the cytoplasm. Compared with model group, the expression levels of IL-1β and IL-6 in LPS+DHLX group were significantly decreased (P<0.05), the expression level of TAK1 was increased and the expression level of TRAF6 was decreased (P<0.05), the content of TAK1-TRAF6 protein complex was significantly increased (P<0.01), and the two proteins were significantly co-located in cytoplasm. Compared with LPS+DHLX group, the expression levels of IL-1β and IL-6 in other groups were significantly decreased (P<0.05,P<0.01). TAK1-TRAF6 protein complex content in each group was significantly decreased after pathway blocker intervention (P<0.05), while TAK1-TRAF6 protein complex content in each group was significantly increased after pathway blocker combined with DHLX intervention (P<0.05). Co-localization of the TAK1-TRAF6 in cytoplasm was not obvious. ConclusionIn the LPS-induced inflammatory response of bile duct cells, the binding of TAK1 and TRAF6 showed a weakening trend, but DHLX could reverse the phenomenon, we think the mechanism of action may be related to promoting the mutual binding of TAK1 and TARF6 to inhibit the activation of the NF-κB/MAPK signaling pathway.

6.
Article in Chinese | WPRIM | ID: wpr-940393

ABSTRACT

ObjectiveTo observe the therapeutic effects of the combined therapy of lung and intestine, a common treatment for pulmonary diseases in traditional Chinese medicine (TCM), on bronchial asthma mice, and further detect the changes of vasoactive intestinal peptide (VIP) and p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway-related proteins which are closely related to the pathogenesis of asthma, in order to elucidate the mechanism of the combined therapy of lung and intestine in the treatment of bronchial asthma. MethodA total of 60 Kunming mice were randomly divided into normal group, model group, dexamethasone group (0.5 mg·kg-1·d-1), TCM group (2.73 g·kg-1·d-1), and lung-intestine treatment group (6.825 g·kg-1·d-1), 12 mice in each group. All mice except the normal group were sensitized by ovalbumin to induce bronchial asthma. After 30 days of intragastric administration, serum and lung tissue samples were obtained. The content of VIP, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the serum of mice in each group was detected by enzyme-linked immunosorbent assay (ELISA). The mRNA levels of TNF-α, IL-6, and p38 MAPK in lung tissues of mice were detected by real-time quantitative polymerase chain reaction (Real-time PCR), and the protein levels of TNF-α, IL-6, p38 MAPK, and phosphorylated p38 MAPK (p-p38 MAPK) in lung tissues of mice were assayed by Western blot (WB). ResultCompared with the normal group, the model group showed decreased content of serum VIP (P<0.05), increased content of TNF-α and IL-6 (P<0.05), up-regulated mRNA levels of TNF-α, IL-6, and p38 MAPK, and elevated protein levels of TNF-α, IL-6, and p-p38 MAPK/p38 MAPK in lung tissues (P<0.05). Compared with the model group, the treatment groups exhibited increased content of serum VIP, TNF-α, and IL-6 (P<0.05), down-regulated mRNA levels of TNF-α, IL-6, and p38 MAPK, and lower protein levels of TNF-α, IL-6, and p-p38 MAPK/p38 MAPK in lung tissues (P<0.05). As compared with the lung-intestine treatment group, the serum TNF-α and IL-6 levels in the dexamethasone group were increased (P<0.05), and the mRNA and protein levels of TNF-α and IL-6 in lung tissues were down-regulated (P<0.05), while the levels of p38 MAPK, VIP mRNA, and p-p38 MAPK/p38 MAPK protein in lung tissues were up-regulated (P<0.05). The serum VIP, TNF-α, and IL-6 levels in the TCM group were decreased (P<0.05), and the mRNA levels of TNF-α, IL-6, p38 MAPK and protein levels of TNF-α, IL-6, p-p38 MAPK/p38 MAPK in lung tissues were up-regulated (P<0.05), while the level of VIP mRNA in lung tissues was down-regulated (P<0.05). ConclusionThrough increasing endogenous VIP and inhibiting the excessive activation of p38 MAPK signaling pathway, the combined therapy of lung and intestine can reduce the release of inflammatory factors, inhibit pulmonary inflammation response, and treat bronchial asthma.

7.
Article in Chinese | WPRIM | ID: wpr-940290

ABSTRACT

ObjectiveProteoglycan TPG-1 isolated from Trametes robiniophila(Huaier) has proved to have anti-hepatoma activity, and this paper aims to explore the molecular mechanism. MethodHuman hepatoma SK-HEP-1 cells were treated with TPG-1 (0, 0.05, 0.1, 0.25, 0.5, 1 g·L-1). Then cell survival was detected by methyl thiazolyl tetrazolium (MTT) and apoptosis by flow cytometry. In addition, expression of genes in SK-HEP-1 cells treated with or without TPG-1 was examined by DNA microarray to preliminarily explore the anti-hepatoma molecular mechanism of TPG-1. ResultTPG-1 inhibited the proliferation of SK-HEP-1 cells as compared with the blank group (P<0.01). After treatment with 1 g·L-1 TPG-1 for 48 h, the apoptosis rate of SK-HEP-1 cells increased (P<0.01), and TPG-1 promoted the cleavage of cysteinyl aspartate specific proteinase (Caspase)-3 and Caspase-7, the key mediators of apoptosis (P<0.01). Additionally, TPG-1 (1 g·L-1) suppressed the migration of SK-HEP-1 cells (P<0.05). A total of 971 differentially expressed genes (DEGs) were identified in SK-HEP-1 cells after treatment with TPG-1, with 486 up-regulated and 485 down-regulated. These DEGs were mainly involved in the Gene Ontology (GO) terms of interleukin-6 (IL-6) biosynthesis, antigen processing and presentation, superoxide dismutase activity, positive regulation of mitogen-activated protein kinase kinase kinase (MAPKKK) cascade, nature killer (NK) cell chemotaxis, and chemokine biosynthesis, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of nucleotide-binding oligomerization domain (NOD)-like receptor signaling pathway, apoptosis, Toll-like receptor signaling pathway, retinoic acid-inducible gene-Ⅰ (RIG-Ⅰ)-like receptor signaling pathway, T-cell receptor signaling pathway, and chemokine signaling pathway. Western blot results showed that TPG-1 (1 g·L-1) activated mitogen-activated protein kinase (MAPK) signaling pathway in SK-HEP-1 cells (P<0.01). ConclusionProteoglycan TPG-1 inhibited the proliferation and migration, and induced apoptosis of human hepatoma SK-HEP-1 cells. Up-regulation of MAPK signaling pathway may be responsible for the growth inhibition of human hepatoma SK-HEP-1 cells by TPG-1.

8.
Article in Chinese | WPRIM | ID: wpr-940180

ABSTRACT

ObjectiveTo study the effect of apigenin on the proliferation and apoptosis of human colon cancer CL187 cells and the underlying mechanisms. MethodHuman colorectal cancer CL187 cells were treated with different concentrations of apigenin (0, 30, 45, 60 mg·L-1) according to the results of the preliminary experiment. The proliferation of CL187 cells was detected by methyl thiazolyl tetrazolium (MTT) and colony formation assays, and the apoptosis was observed via Hoechst 33258 staining. Real-time fluorescence quantitative PCR was conducted to determine the mRNA levels of cysteine protease-3 (Caspase-3), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in the CL187 cells treated with apigenin. Western blot was employed to measure the protein levels of Caspase-3, Bcl-2, and Bax associated with apoptosis, protein kinase B (Akt) and phosphorylated Akt (p-Akt) in phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathway, and extracellular signal-regulated kinases 1/2 (ERK1/2), p-ERK1/2, c-Jun N-terminal kinase (JNK), p-JNK, p38 mitogen-activated protein kinase (MAPK), and p-p38 MAPK protein in MAPK pathway. ResultCompared with the blank group, the apigenin groups had low cell survival rates and high inhibition rates on cell proliferation (P<0.01). Apigenin decreased the cell clone number and clone formation rate, and increased the inhibition rate on clone formation (P<0.01). After CL187 cells were treated with different concentrations of apigenin for 48 h, typical apoptosis characteristics such as nuclear pyknosis, chromatin condensation, and enhanced fluorescence reaction were observed. Compared with blank group, 45, 60 mg·L-1 apigenin treatments down-regulated the mRNA level of anti-apoptotic gene Bcl-2 (P<0.01) and all the apigenin treatments up-regulated those of the pro-apoptotic genes Bax and Caspase-3 (P<0.05, P<0.01). Similarly, apigenin treatments down-regulated the protein level of Bcl-2 (P<0.05, P<0.01) and up-regulated those of Caspase-3 (P<0.05, P<0.01) and Bax (P<0.01, 45, 60 mg·L-1). The blank group had higher protein level of Akt than the 60 mg·L-1 apigenin group (P<0.01), higher protein levels of p-Akt, ERK1/2, and p-ERK1/2 than the 45, 60 mg·L-1 apigenin groups (P<0.01), and higher protein levels of JNK and p-JNK than the apigenin groups (P<0.05, P<0.01). Compared with blank group, 60 mg·L-1 apigenin up-regulated the protein level of p38 MAPK (P<0.05), and all the apigenin groups up-regulated that of p-p38 MAPK (P<0.01). Furthermore, apigenin lowered the p-Akt/Akt ratio (P<0.05, P<0.01) and p-ERK1/2/ERK1/2 ratio (P<0.01), while it increased the p-JNK/JNK ratio (45, 60 mg·L-1; P<0.05, P<0.01) and p-p38 MAPK/p38 MAPK ratio (P<0.05, P<0.01). ConclusionApigenin can inhibit the proliferation and promote the apoptosis of CL187 cells by inhibiting the PI3K/Akt signaling pathway and regulating the expression of proteins in the MAPK signaling pathway.

9.
Article in Chinese | WPRIM | ID: wpr-940148

ABSTRACT

ObjectiveTo study the effect of apigenin on the proliferation and apoptosis of human colon cancer CL187 cells and the underlying mechanisms. MethodHuman colorectal cancer CL187 cells were treated with different concentrations of apigenin (0, 30, 45, 60 mg·L-1) according to the results of the preliminary experiment. The proliferation of CL187 cells was detected by methyl thiazolyl tetrazolium (MTT) and colony formation assays, and the apoptosis was observed via Hoechst 33258 staining. Real-time fluorescence quantitative PCR was conducted to determine the mRNA levels of cysteine protease-3 (Caspase-3), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in the CL187 cells treated with apigenin. Western blot was employed to measure the protein levels of Caspase-3, Bcl-2, and Bax associated with apoptosis, protein kinase B (Akt) and phosphorylated Akt (p-Akt) in phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathway, and extracellular signal-regulated kinases 1/2 (ERK1/2), p-ERK1/2, c-Jun N-terminal kinase (JNK), p-JNK, p38 mitogen-activated protein kinase (MAPK), and p-p38 MAPK protein in MAPK pathway. ResultCompared with the blank group, the apigenin groups had low cell survival rates and high inhibition rates on cell proliferation (P<0.01). Apigenin decreased the cell clone number and clone formation rate, and increased the inhibition rate on clone formation (P<0.01). After CL187 cells were treated with different concentrations of apigenin for 48 h, typical apoptosis characteristics such as nuclear pyknosis, chromatin condensation, and enhanced fluorescence reaction were observed. Compared with blank group, 45, 60 mg·L-1 apigenin treatments down-regulated the mRNA level of anti-apoptotic gene Bcl-2 (P<0.01) and all the apigenin treatments up-regulated those of the pro-apoptotic genes Bax and Caspase-3 (P<0.05, P<0.01). Similarly, apigenin treatments down-regulated the protein level of Bcl-2 (P<0.05, P<0.01) and up-regulated those of Caspase-3 (P<0.05, P<0.01) and Bax (P<0.01, 45, 60 mg·L-1). The blank group had higher protein level of Akt than the 60 mg·L-1 apigenin group (P<0.01), higher protein levels of p-Akt, ERK1/2, and p-ERK1/2 than the 45, 60 mg·L-1 apigenin groups (P<0.01), and higher protein levels of JNK and p-JNK than the apigenin groups (P<0.05, P<0.01). Compared with blank group, 60 mg·L-1 apigenin up-regulated the protein level of p38 MAPK (P<0.05), and all the apigenin groups up-regulated that of p-p38 MAPK (P<0.01). Furthermore, apigenin lowered the p-Akt/Akt ratio (P<0.05, P<0.01) and p-ERK1/2/ERK1/2 ratio (P<0.01), while it increased the p-JNK/JNK ratio (45, 60 mg·L-1; P<0.05, P<0.01) and p-p38 MAPK/p38 MAPK ratio (P<0.05, P<0.01). ConclusionApigenin can inhibit the proliferation and promote the apoptosis of CL187 cells by inhibiting the PI3K/Akt signaling pathway and regulating the expression of proteins in the MAPK signaling pathway.

10.
Article in Chinese | WPRIM | ID: wpr-876051

ABSTRACT

@#[Abstract] Objective: To explore the effect and mechanism of splicing factor 3b subunit 6 (SF3b6) on the proliferation, apoptosis, invasion and migration of gastric cancer cells. Methods: Tissue microarrays were used to detect the expression of SF3b6 in gastric cancer tissues and adjacent tissues. WB and qPCR were used to detect the expression of SF3b6 in normal immortalized gastric epithelial cells (GES-1) and gastric cancer cell lines (HGC27, AGS, BGC823, MGC803, SGC7901, MKN45). AGS and MGC803 cells were transfected with SF3b6 siRNA, and BGC823 and SGC7901 cells were transfected with SF3b6 over-expression plasmid for functional experiments. CCK-8 assay was used to detect the regulation of SF3b6 on the proliferation of gastric cancer cells; Transwell migration and invasion experiments were used to detect the effect of SF3b6 on the migration and invasion of gastric cancer cells; Flow cytometry was used to detect cell apoptosis; and WB was adopted to detect expressions of apoptosis and migration-related molecules and MAPK signaling pathway associated proteins. Results: The expression level of SF3b6 in gastric cancer MGC803 and AGS cells was significantly higher while in BGC823 and SGC7901 cells was significantly lower than that in normal gastric epithelial GES-1 cells (P<0.05 or P<0.01). SF3b6 knockdown inhibited the proliferation, migration and invasion, but promoted cell apoptosis of gastric cancer cell lines AGS and MGC803 (P<0.05 or P<0.01); However, over-expression of SF3b6 promoted the proliferation, migration and invasion of gastric cancer cell lines BGC823 and SGC7901 (P<0.05 or P<0.01). Mechanism study showed that SF3b6 knockdown promoted the activation of JNK and p38 and expression of apoptosis-related protein cleaved caspase-9, cleaved PARP and Bax (P<0.05 or P<0.01), and meanwhile inhibited the process of epithelial to mesenchymal transition (EMT) in gastric cancer cells. Conclusion: The splicing factor SF3b6 enhances cell proliferation and migration via MAPK signaling pathway, thereby promoting tumor progression.

11.
Article in Chinese | WPRIM | ID: wpr-906126

ABSTRACT

Objective:To investigate the protective effect of quercetin (Qu) on articular cartilage of knee osteoarthritis and its mechanism by inhibiting p38 mitogen activated protein kinase (MAPK) signaling pathway. Method:Through the network pharmacology technology,we scientifically predicted and analyzed the target factors and signal pathways of Qu in the protection of articular cartilage in patients with osteoarthritis. We selected a prediction pathway closely related to osteoarthritis and validated it by cell experiment <italic>in vitro</italic>. The best intervention concentration of the drug was selected by cell counting kit-8 (CCK-8) method. The osteoarthritis and its closely related inflammatory factors interleukin(IL)-1<italic>β</italic> and tumor necrosis factor(TNF)-<italic>α</italic> were detected by enzyme linked immunosorbent assay(ELISA). The expression of related mRNA and protein in p38 signal pathway after Qu intervention were detected by quantitative real time polymerase chain reaction(Real-time PCR) and Western blot. Result:It was predicted that MAPK signal pathway was closely related to osteoarthritis by network pharmacology,and p38 MAPK pathway,which was most closely related to osteoarthritis,was selected for study. The results showed that 100 μmol·L<sup>-1</sup> Qu had the most obvious effect in decreasing the expression of related inflammatory factors,inhibited the expression of p38,phosphorylated(p)-p38,matrix metalloproteinase-13(MMP-13),A disintegrin-like and metalloproteinase with thrombospondin type-1 motifs-4(ADAMTS-4) in the pathway,and promoted the expression of CollagenⅡ. Conclusion:Qu could decrease the expression of cartilage inflammatory factors in the prevention and treatment of osteoarthritis,and the effect can be well developed by intervening and inhibiting p38 MAPK pathway related factor expression level. All the results show that Qu can decrease osteoarthritis inflammatory factors and protect articular cartilage in patients with osteoarthritis.

12.
Article in Chinese | WPRIM | ID: wpr-921649

ABSTRACT

The present study observed the effect of Guanxin Zhitong Capsules(GXZT) on the lipotoxicity of vascular endothelial cells and investigated the mechanism of GXZT in atherosclerosis treatment. The lipotoxicity model in human umbilical vein endothelial cells(HUVECs) was induced by palmitic acid(PA) stimulation. These cells were divided into a normal control group(NC, 15% normal serum), a model group(PA, 0.6 mmol·L~(-1) PA+15% normal serum), a high-dose GXZT group(GXZT-H, 0.6 mmol·L~(-1) PA+15% GXZT-medicated serum), a medium-dose GXZT group(GXZT-M, 0.6 mmol·L~(-1) PA+10% GXZT-medicated serum+5% normal serum) and a low-dose GXZT group(GXZT-L, 0.6 mmol·L~(-1) PA+5% GXZT-medicated serum+10% normal serum). HUVECs were detected for cell viability by cell counting kit-8(CCK-8) assay, apoptosis by flow cytometry, mitochondrial membrane potential(MMP) by JC-1 labeled laser scanning confocal microscopy, and total and phosphorylated proteins of p38, ERK1/2, and JNK1/2 in the mitogen-activated protein kinases(MAPK) signaling pathway by Western blot. The phosphorylated level was calcula-ted. Compared with the NC group, the PA group showed decreased cell viability and MMP(P<0.01, P<0.01), elevated apoptosis(P<0.01), and up-regulated phosphorylated levels of p38, ERK1/2, and JNK1/2(P<0.01, P<0.01, P<0.01). Compared with the PA group, the GXZT-H, GXZT-M, and GXZT-L groups showed increased cell viability and MMP(P<0.01, P<0.01, P<0.01), reduced apoptosis(P<0.01), and down-regulated protein expression and phosphorylated levels of p38, ERK1/2 and JNK1/2 in the MAPK signaling pathway(P<0.01, P<0.01, P<0.01). In conclusion, the results suggest that GXZT functions via blocking MAPK signaling pathway to relieve the damage of HUVECs induced by PA.


Subject(s)
Apoptosis , Capsules , Human Umbilical Vein Endothelial Cells/metabolism , Humans , MAP Kinase Signaling System , Palmitic Acid/toxicity , Signal Transduction , p38 Mitogen-Activated Protein Kinases/metabolism
13.
Acupuncture Research ; (6): 299-304, 2020.
Article in Chinese | WPRIM | ID: wpr-844171

ABSTRACT

OBJECTIVE: To observe the effect of electroacupuncture (EA) on degranulation of intraperitoneal mast cells (MCs) and expression of mitogen-activated protein kinase (MAPK) signaling related proteins, tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in urticaria rats, so as to reveal its mechanisms underlying improvement of urticaria. METHODS: Thirty-two SD rats were randomly divided into control,model,EA and medication groups (n=8 in each group). The urticaria model was established by using passive cutaneous anaphylaxis (PCA) reaction method. EA (2 Hz /15 Hz, 1 mA) was applied to bilateral "Zusanli"(ST36), "Quchi "(LI11) and "Xuehai"(SP10) for 20 min,once daily for 7 consecutive days before antigen attack. Rats of the medication group were treated by gavage of Loratadine(1 mg•kg-1•d-1)for 7 days. The diameter of cutaneous Evan's blue spots was measured to evaluate the severity of PCA. Intraperitoneal fluid smears were prepared to observe the degranulation state of MCs. The contents of TNF-α and IL-6 in the intraperitoneal fluid were detected by ELISA, and the expression of extracellular signal-regulated kinase (ERK), phosphorylated (p)-ERK, c-Jun N-terminal kinase (JNK), p-JNK, P38MAPK and p-P38MAPK of the acquired intraperitoneal MCs was detected by Western blot. RESULTS: The diameter of cutaneous Evan's blue spot was significantly increased in the model group than that in the control group (P<0.01), and considerably decreased in both EA and medication groups compared with the model group(P<0.01). After modeling,the percentage of degranulated MCs, contents of TNF-α and IL-6, and expression levels of ERK, p-ERK, JNK, p-JNK, P38MAPK and p-P38MAPK were remarkably increased in the mo-del group than those in the control group (P<0.01, P<0.05). After the treatment, the percentage of degranulated MCs, contents of TNF-α and IL-6, and expression levels of p-ERK, JNK, p-JNK and p-P38MAPK were obviously decreased in both EA and medication groups relevant to the model group (P<0.01, P<0.05), while no significant changes were found in the expression of ERK in both EA and medication groups, and P38MAPK in the EA group. Compared with the model and EA groups, expression levels of P38MAPK were down-regulated in the medication group (P<0.05). CONCLUSION: EA can reduce skin allergic reaction in rats with urticaria, which may be related to its effects in inhibiting the degranulation of intraperitoneal MCs, down-regulating the expression of MAPK signaling-related proteins and the level of pro-inflammatory factors TNF-α and IL-6 in intraperitoneal MCs.

14.
Article in Chinese | WPRIM | ID: wpr-873077

ABSTRACT

Objective::To explore the effect of Tongxie Yaofang on p38 mitogen activated protease(p38 MAPK), mitogen and stress protein kinase 1(MSK1), cyclic adenosine effector response element binding protein(CREB)mRNA and protein expression in colon tissue of diarrhea type irritable bowel syndrome (D-IBS) rat model with liver depression and spleen deficiency(GYPX), and interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), and the content of total superoxide dismutase (T-SOD) and malondialdehyde (MDA) in serum. Method::The 60 SPF Wistar rats were randomly divided into 6 groups, with 10 rats in each group. Except the normal group, rats in the other groups were given by gavage with folium sennae and chronic bondage to establish D-IBS with GYPX for 14 days. The low, medium, and high doses Tongxie Yaofang were administered to Tongxie Yaofang(2.25, 4.5, 9 g·kg-1)gavage respectively. The piveronium bromide group was given piveronium bromide tablets suspension(0.02 g·kg-1)gavage.The normal group group and model group were given the same volume normal saline for 21 days. After the last gavage for 18 hours, the heart blood was collected and the colon tissue was dissected. Real-time PCR was used to observe the expression of p38 MAPK, MSK1 and CREB mRNA in rat colon. Western blot was used to observe the expression of p38 MAPK, MSK1 and CREB protein. ELISA was used to observe the contents of IL-1β, IL-6 and TNF-α in colon. Hydroxylamine was used to observe the T-SOD level in serum, thiobarbituric acid(TBA)was used to observe the MDA content in serum. hematoxyl in-eosin(HE) staining was used to observe the morphological changes of colon tissues. Result::Compared with normal group, the expression of p38 MAPK, MSK1, CREB mRNA and the protein content of p38 MAPK, MSK1 and CREB in the colon tissue of model group rats increased significantly, while the content of IL-1β, IL-6 and TNF-α increased significantly(P<0.05). The level of serum T-SOD decreased significantly, and the content of MDA increased significantly(P<0.05). Compared with the model group, the medium and high dose group of Tongxie Yaofang significantly decreased the expression of p38 MAPK mRNA, content of p38 MAPK, CREB protein and IL-1β, IL-6, TNF-α in colon tissue(P<0.05). The level of serum T-SOD increased significantly, and the content of MDA decreased significantly(P<0.05). High dose group of Tongxie Yaofang can significantly decreased the expression of MSK1, CREB mRNA, content of MSK1 protein(P<0.05). Histopathological observation showed that no significant organic lesions were observed in the colonic morphology of each group of rats, which was consistent with the morphological characteristics of IBS. Conclusion::Tongxie Yaofang has a significant dose-effect relationship in the treatment of D-IBS rats with GYPX in a certain range, which may be related to its increases antioxidant stress and inhibit activation of p38 MAPK signaling pathway and reducing the level of downstream inflammatory factors.

15.
Article in Chinese | WPRIM | ID: wpr-872651

ABSTRACT

Objective:To study the effects of licochalcone A (LCA) on the proliferation and apoptosis of rheumatoid arthritis fibroblast-like synoviocytes (MH7A) as well as the related inflammatory factors, also to reveal the relevance between mitogen activated protein kinase (MAPK) signaling pathway and LCA regulation of MH7A cell proliferation and apoptosis. Method:MH7A cells were cultured and divided into blank group, LCA groups (10,20,40 μmol·L-1). The proliferation of MH7A cells was detected by methylthiazolyldiphenyl-tetrazolium bromide(MTT)and immunofluorescence staining. The cell cycle of MH7A cells was determined by flow cytometry after PI staining and apoptosis was detected by flow cytometry after Annexin V/PI staining. The effect of LCA on interleukin-1β(IL-1β) mRNA was detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). Western blot was used to detect the effect of LCA on the key proteins of MAPK signaling pathway, meanwhile, PD98059, a specific ERK inhibitor, was used to observe the expression levels of p-ERK and IL-1β. Result:Compared with blank group, LCA could inhibit the proliferation of MH7A cells in a dose-dependent manner, and the number of living cells decreased significantly(P<0.01), while the number of early apoptotic cells increased significantly(P<0.01). Compared with the tumor necrosis factor-α (TNF-α,10 μg·L-1)group, LCA could reverse the expression of IL-1β mRNA induced by TNF-α(P<0.01). and compared with the blank group, LCA also promoted the phosphorylation of ERK, JNK and p38 in a dose-dependent manner(P<0.01). After ERK inhibitor PD98059 inhibited ERK phosphorylation, the inhibitory effect of LCA 10, 20 μmol·L-1 on IL-1β disappeared. Conclusion:LCA can inhibit the proliferation and induce apoptosis of MH7A cells, which may be related to the phosphorylation of MAPK pathway related proteins, and then inhibit the expression of inflammatory factors.

16.
Article in Chinese | WPRIM | ID: wpr-841557

ABSTRACT

Objective: To observe the improvement effect of panax notoginseng saponins on the cardiac function of the rats with chronic heart failure (CHF), and to investigate the possible mechanism. Methods: Abdominal aortic constriction was used to establish the CHF rat models. Sixty model rats were randomly divided into model group, positive control group, low, medium and high doses of panax notoginseng saponins groups ( n= 12). Another 12 rats were taken as sham operation group. Color floppier ultrasound was used to detect the left ventricular end-diastolic diameter (LVEDD)» left ventricular end-systolic diameter (LVESD), left ventricular posterior wall diastolic thickness (LVPWD), left ventricular ejection fraction (LVEF), cardiac output (CO) , left ventricular end-diastolic pressure (LVEDP), left ventricular systolic pressure (LVSP), maximal rate of increase of left ventricular pressure (+dp/dt max) and maximal rate of decrease of left ventricular pressure ( dp/dt max) of the rats in various groups∗ HE staining was used to observe the pathomorphology of myocardium tissue of the rats in various groups∗ TUNEL method was used to detect the apoptotic rates of cadiomyocytes of the rats in various groups∗ and Western blotting method was used to detect the expression levels of extracellular signal-regulated kinase (ERK), phosphorylation ERK (p-ERK), c-Jun NH2-terminal protein kinase (JNK), phosphorylation JNK (p-JNK), p38, and p-p38 proteins in myocardium tissue of the rats in various groups. ELISA was used to determine the serum levels of tumor necrosis factor-a (TNF-a) and interleukin-6 (IL-6) of the rats in various groups. Results: Compared with sham operation group, the LVEDI), LVESD, LVPWD, LVEDP and dp/dt max of the rats in model group were significantly increased (P

17.
Article in Chinese | WPRIM | ID: wpr-828932

ABSTRACT

OBJECTIVE@#To investigate the effect of periostin on hypoxia-induced oxidative stress and apoptosis in human periodontal ligament fibroblasts and the molecular mechanism involved.@*METHODS@# cultured human periodontal ligament fibroblasts were placed in an anaerobic gas-producing bag for hypoxia treatment for 48 h followed by treatment with periostin at low (25 ng/mL), moderate (50 ng/mL) or high (100 ng/mL) doses. MTT assay was used to measure the cell viability, and the cell apoptosis rate was determined using flow cytometry. The contents of IL-1β, IL-6 and TNF-α in the cells were determined with ELISA, and ROS levels were measured using a fluorescent plate reader. The intracellular SOD activity was detected using ELISA. The expressions of HIF-1α, P21, cyclin D1, Bax, cleaved caspase-3, Bcl-2, P38MAPK and p-p38 MAPK proteins in the cells were detected with Western blotting.@*RESULTS@#Hypoxia treatment significantly reduced the cell viability ( < 0.05), increased P21, Bax, and cleaved caspase-3 protein levels ( < 0.05), promoted cell apoptosis ( < 0.05), and decreased cyclin D1 and Bcl-2 protein levels ( < 0.05) in the cells. Compared with the hypoxic group, the cells treated with periostin at different concentrations showed significantly increased cell viability ( < 0.05) with significantly lowered apoptotic rates ( < 0.05) and decreased expression levels of Bax and cleaved caspase-3 ( < 0.05) but significantly increased expression levels of cyclin D1 and Bcl-2 ( < 0.05). Hypoxic exposure of the cells resulted in significantly increased expression levels of HIF-1α and p-p38 MAPK ( < 0.05) and increased levels of IL-1β, IL-6, TNF-α and ROS ( < 0.05) but decreased SOD activity ( < 0.05). Periostin treatment at different concentrations significantly lowered the expression levels of HIF-1α and p-p38 MAPK ( < 0.05) and the levels of IL-1β, IL-6, TNF-α and ROS ( < 0.05) and significantly increased SOD activity in the hypoxic cells ( < 0.05).@*CONCLUSIONS@#Periostin promotes the proliferation, inhibits apoptosis, enhances cellular antioxidant capacity, and reduces inflammatory damage in human periodontal ligament fibroblasts exposed to hypoxia possibly by inhibiting the activation of the p38 MAPK signaling pathway.


Subject(s)
Apoptosis , Fibroblasts , Humans , Hypoxia , Oxidative Stress , Periodontal Ligament , Signal Transduction , p38 Mitogen-Activated Protein Kinases
18.
Article in Chinese | WPRIM | ID: wpr-818378

ABSTRACT

Objective Melanoma is a malignant tumor with high clinical malignancy, rapid progression, poor prognosis and high mortality. It is of great clinical significance to explore the molecular mechanism of high metastasis of melanoma and find new tumor markers and therapeutic targets. The expression of mitochondrial fusion protein-2 (MFN2) in cutaneous melanoma tissues and peritumoral tissues was studied, and its correlation with clinical parameters was analyzed. The correlation between MFN2 and the biological behavior of melanoma was also discussed. Methods Immunohistochemistry was used to detect the expression of Med19 protein in cutaneous melanoma tissues and peritumoral tissues, and to analyze the correlation between MFN2 expression and clinical factors in patients. The lentiviral vector containing MFN2 coding sequence (Lenti-MFN2) infected with melanoma cell B16 was set up as the study group, and the lentivirus with green fluorescent protein gene (Lenti-GFP) as the control group. The mRNA and protein expressions of MFN2 and Ras-Raf1-ERK1/2 signaling pathways in transfected cells were detected by real-time quantitative PCR (qRT-PCR) and Western blot. Flow cytometry was used to detect changes in cell cycle and apoptosis. MTT assay and colony formation assay were used to detect changes in cell viability and proliferation capacity. Results The positive expression rate of MFN2 in peritumoral tissues was significantly higher than that in the skin melanoma tissues (83.9% vs 30.6%, P<0.05). MFN2 expression was associated with both lymph node metastasis and TNM staging (P<0.05). Compared with the control group, the proliferation activity and colony forming ability of B16 cells in the study group were significantly inhibited (P<0.05); the proportion of G0/G1 phase cells in B16 cells in the study group significantly increased [(46.3±10.3)% vs (67.9±12.6)%], and the apoptosis rate significantly increased [(12.5±1.6)% vs (57.4±12.6)%], with statistically significant differences (P<0.05). In comparison with the control group, the expression of Ras, Raf, ERK1/2 mRNA and protein in the study group significantly decreased (P<0.05). Conclusion MFN2 is down-regulated in melanoma tissues. Up-regulation of MFN2 expression can inhibit melanoma proliferation and promote cell apoptosis, which may be related to the inhibition of Ras-MAPK signaling pathway activity.

19.
Article in Chinese | WPRIM | ID: wpr-802532

ABSTRACT

Objective: To investigate the expression changes of mitogen activated protein kinase(MAPK) signaling pathway-related genes in ApoE-/- mice and the intervention effect of Huayu Qutan recipe on them, in order to explore the anti-atherosclerotic(AS) effect and possible mechanism of Huayu Qutan recipe. Method: Fifty ApoE-/- mice were randomly divided into model group, tanshinone ⅡA group (30 mg·kg-1·d-1), and high-dose phlegm group (20 g·kg-1·d-1), the middle-dose group (10 g·kg-1·d-1), and the low-dose group (5 g·kg-1·d-1), and 10 C57BL/6/J mice were included in the blank controls group. Automated biochemical analyzer was used to detect serum triglyceride(TG), cholesterol(TC), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C) content; hematoxylin-eosin(HE) was used to detect aortic plaque in each group; oil red O was used to detect liver lipid deposition in each group; enzyme-linked immuno sorbent assay(ELISA) was used to determine vascular cell adhesion protein-1(VCAM-1), intercellular adhesion molecule-1(ICAM-1), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α) in serum of each group; Western blot was performed to detect c-Jun N-terminal kinase (JNK), phosphorylated c-Jun N-terminal kinase(p-JNK), c-Jun N-terminal kinase(JNK), p38 MAPK, p-p38 MAPK, extracellular regulated protein kinases (ERK), and expression of p-ERK. Result: Compared with the blank control group, serum TC, TG, LDL-C levels in the model group were significantly increased, while HDL-C levels were significantly decreased; aortic plaques were observed in the aortic lumen, and lots of lipid deposition were observed in the liver cells. Serum inflammatory factors TNF-α, IL-6, VCAM-1, ICAM-1 increased(PPα, IL-6, VCAM-1 and ICAM-1 were decreased. The expressions of p-p38 MAPK and p-JNK genes in intravascular associated MAPK signaling pathway were significantly decreased. p-ERK expression trend was not obvious(PConclusion: Huayu Recipe can inhibit the formation of aortic plaque in ApoE-/- mice, and its mechanism may be related to the regulation of vascular MAPK signaling pathway gene expression.

20.
Article in Chinese | WPRIM | ID: wpr-802232

ABSTRACT

Objective: To investigate the protective effect of Perillae Folium with aqueous extract (PFAE) on some key factors of Adriamycin (ADR)-induced oxidative injury in human renal tubular epithelial cells(HK-2), including the survival rate, oxidative injury indexes and cell apoptosis,in order to define the underlying mechanism. Method: A model of ADR-induced HK-2 cells oxidative injury was established in vitro, then cell viability was detected by cell counting kit-8 (CCK-8) after intervention with positive reference N-acetylcysteine (NAC) or PFAE (5,15,45 g·L-1) at different concentrations. According to the morphological changes under microscopy, the optimum concentration of PFAE was screened out for the follow-up experiments. Then, the experiments were divided into six groups:blank group, ADR (0.05 g·L-1) group, PFAE (15 g·L-1) group, ADR+PFAE (0.05+15) g·L-1 group, NAC (0.81 g·L-1) group, and ADR+NAC (0.05+81) g·L-1 group. After that, malondialdehyde (MDA), superoxide dismutase (SOD), total antioxidant capacity(TAC) were measured in the cell homogenate after 24 h administration. The level of reactive oxygen species (ROS) was detected by 2',7'-dichloroflurescin diacetate (DCFH-DA) fluorescence probe. Flow cytometry and TdT-mediated dUTP Nick-End Labeling (TUNEL) were used to monitor the cell apoptosis. Western blot was used to observed the expressions of mitochondrial apoptosis-associated proteins, like B lymphocyte tumor-2 gene (Bcl-2), Bcl-2 related X protein (Bax), cysteine aspartate protease-9 (Caspase-9), cysteine aspartate protease-3 (Caspase-3) and poly ADP-ribose polymerase (PARP), as well as their shear bodies. In addition, the phosphorylation protein expressions of p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK), c-Jun amino-terminal kinase (JNK) in mitogen-activated protein kinase (MAPK) signaling transduction pathway were detected by Western blot. Result: Compared with blank group, ADR group showed a decreased cell viability (PPPPPPPP-1. The ATC and SOD levels were increased in ADR+PFAE group and ADR+NAC group (PPConclusion: PFAE could alleviate the oxidative injury of HK-2 cells induced by ADR, and have an antioxidant effect, which inhibited cell apoptosis through mitochondrial apoptotic pathway and ERK/p38 MAPK signaling pathway.

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