Mannitol inhibits the proliferation of neural stem cell by a p38 mitogen-activated protein kinase-dependent signaling pathway / 中华创伤杂志(英文版)

PURPOSE@#Mannitol is one of the first-line drugs for reducing cerebral edema through increasing the extracellular osmotic pressure. However, long-term administration of mannitol in the treatment of cerebral edema triggers damage to neurons and astrocytes. Given that neural stem cell (NSC) is a subpopulation of main regenerative cells in the central nervous system after injury, the effect of mannitol on NSC is still elusive. The present study aims to elucidate the role of mannitol in NSC proliferation.@*METHODS@#C57 mice were derived from the animal house of Zunyi Medical University. A total of 15 pregnant mice were employed for the purpose of isolating NSCs in this investigation. Initially, mouse primary NSCs were isolated from the embryonic cortex of mice and subsequently identified through immunofluorescence staining. In order to investigate the impact of mannitol on NSC proliferation, both cell counting kit-8 assays and neurospheres formation assays were conducted. The in vitro effects of mannitol were examined at various doses and time points. In order to elucidate the role of Aquaporin 4 (AQP4) in the suppressive effect of mannitol on NSC proliferation, various assays including reverse transcription polymerase chain reaction, western blotting, and immunocytochemistry were conducted on control and mannitol-treated groups. Additionally, the phosphorylated p38 (p-p38) was examined to explore the potential mechanism underlying the inhibitory effect of mannitol on NSC proliferation. Finally, to further confirm the involvement of the p38 mitogen-activated protein kinase-dependent (MAPK) signaling pathway in the observed inhibition of NSC proliferation by mannitol, SB203580 was employed. All data were analyzed using SPSS 20.0 software (SPSS, Inc., Chicago, IL). The statistical analysis among multiple comparisons was performed using one-way analysis of variance (ANOVA), followed by Turkey's post hoc test in case of the data following a normal distribution using a Shapiro-Wilk normality test. Comparisons between 2 groups were determined using Student's t-test, if the data exhibited a normal distribution using a Shapiro-Wilk normality test. Meanwhile, data were shown as median and interquartile range and analyzed using the Mann-Whitney U test, if the data failed the normality test. A p < 0.05 was considered as significant difference.@*RESULTS@#Primary NSC were isolated from the mice, and the characteristics were identified using immunostaining analysis. Thereafter, the results indicated that mannitol held the capability of inhibiting NSC proliferation in a dose-dependent and time-dependent manner using cell counting kit-8, neurospheres formation, and immunostaining of Nestin and Ki67 assays. During the process of mannitol suppressing NSC proliferation, the expression of AQP4 mRNA and protein was downregulated, while the gene expression of p-p38 was elevated by reverse transcription polymerase chain reaction, immunostaining, and western blotting assays. Subsequently, the administration of SB203580, one of the p38 MAPK signaling pathway inhibitors, partially abrogated this inhibitory effect resulting from mannitol, supporting the fact that the p38 MAPK signaling pathway participated in curbing NSC proliferation induced by mannitol.@*CONCLUSIONS@#Mannitol inhibits NSC proliferation through downregulating AQP4, while upregulating the expression of p-p38 MAPK.

Mechanism of Stemona tuberosa Alkaloids in Inhibiting Proliferation and Inducing Apoptosis of Non-small Cell Lung Cancer NCI-H460 Cells / 中国实验方剂学杂志

ObjectiveTo study the effect and underlying mechanism of Stemona tuberosa alkaloids on the proliferation and apoptosis of human non-small cell lung cancer NCI-H460 cells. MethodNon-small cell lung cancer NCI-H460 cells were divided into a blank group and S. tuberosa alkaloids groups （50， 100， 150， 200， and 250 mg·L-1）. The effect of S. tuberosa alkaloids on the proliferation of human NCI-H460 cells was observed by thiazolyl blue tetrazolium bromide （MTT） assay and colony formation assay. Cell apoptosis was observed by Hoechst 33258 staining and flow cytometry. Real-time fluorescence-based polymerase chain reaction （Real-time PCR） was used to detect the effect of S. tuberosa alkaloids on the mRNA expression of cysteinyl aspartate-specific protease 3 （Caspase-3）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and epidermal growth factor receptor （EGFR）. The protein expression levels of Caspase-3， Bax， Bcl-2， protein kinase B （Akt）， phosphorylated （p-）Akt， EGFR， c-Jun N-terminal kinase （JNK）， p-JNK， p38 mitogen-activated protein kinase （p38 MAPK）， and p-p38 MAPK were measured by Western blot. ResultCompared with the blank group， the S. tuberosa alkaloids groups showed increased inhibition rate on cell proliferation （P<0.01）， reduced number of cell clones formed and the rate of cell clonal formation （P<0.05， P<0.01）， and increased karyopyknosis， cytoplasmic aggregation， and cell apoptosis rate （P<0.01）. The S. tuberosa alkaloids groups at 100， 150， 200， and 250 mg·L-1 showed increased Caspase-3 mRNA expression （P<0.05）， decreased EGFR mRNA expression （P<0.05， P<0.01）， up-regulated protein expression of Caspase-3 and p-JNK （P<0.01）， and down-regulated protein expression of EGFR and p-Akt （P<0.05， P<0.01）. Additionally， compared with the blank group， the S. tuberosa alkaloids groups showed increased expression of Bax mRNA （P<0.01）， decreased expression of Bcl-2 mRNA （P<0.01）， up-regulated protein expression of Bax and p-p38 MAPK （P<0.01）， and down-regulated protein expression of Bcl-2 （P<0.01）. ConclusionsS. tuberosa alkaloids can inhibit proliferation and induce apoptosis of human non-small cell lung cancer NCI-H460 cells， and the mechanism may be related to the inhibition of EGFR protein expression and phosphorylation of Akt protein， as well as the activation of the JNK/p38 MAPK signaling pathway.

Role of free radicals in cerebral ischemic tolerance of rats induced by limb ischemic preconditioning / 中国病理生理杂志

AIM: To explore whether free radicals participate in cerebral ischemic tolerance and the up-regula-tion of p38 MAPK and ERK signaling pathways in rats induced by limb ischemic preconditioning (LIP). METHODS: A total of 128 Wistar rats with permanent occlusion of bilateral vertebral arteries were randomly divided into sham group (n=16), cerebral ischemia (CI) group (n=16), LIP+CI group (n=16), DMTU (a free radical scavenger)+LIP+CI group (n=64) and DMTU+sham group (n=16). Six rats in each group were used to observe the delayed neuronal death (DND) in hippocampal CA1 region by thionin staining at 7 d after the end of operation. Other 10 rats in each group were used to de-tect the expression of p38 MAPK and ERK in hippocampal CA1 region by immunohistochemistry and Western blot. RE-SULTS: Lethal CI resulted in obvious DND in hippocampal CA1 region. However, LIP reversed the above injurious changes, represented by the decrease in histological grade and the increase in neuronal density compared with CI group (P<0. 01). Moreover, LIP significantly up-regulated the expression of p38 MAPK and ERK in hippocampal CA1 region com-pared with CI group (P<0. 01). Administration of free radical scavenger DMTU via femoral vein before LIP partially re-versed the neuroprotective effect of LIP, and blocked the up-regulation of p38 MAPK and ERK expression in hippocampal CA1 region in rats compared with LIP+CI group (P<0. 01). CONCLUSION: Free radicals are involved in the neuropro-tection and up-regulation of p38 MAPK and ERK expression induced by LIP in rats.

Effect of Stemona tuberosa Alkaloids on Apoptosis and PI3K/Akt and JNK/MAPK Signaling Pathways of Human Lung Cancer A549 Cells / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Stemona tuberosa alkaloids （STA） on apoptosis and phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） and c-Jun N-terminal kinase/p38 mitogen-activated protein kinase （JNK/p38 MAPK） signaling pathways in human lung cancer A549 cells. MethodA549 cells were classified into blank group and STA groups （100， 150， 200， 250， 300 mg⋅L-1）. Thiazole blue （MTT） assay and colony formation assay were used to evaluate the proliferation of A549 cells. Apoptosis was observed based on Hoechst 33258 staining， flow cytometry， and Annexin V-FITC/PI staining. Western blot was employed to detect the expression of apoptosis-related proteins cysteine-aspartic acid protease-3 （Caspase-3）， B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， and Bcl-2， and the expression of PI3K， phosphorylated （p）-PI3K， Akt， p-Akt， JNK， p-JNK， p38 MAPK， and p-p38 MAPK. ResultCompared with the blank group， STA groups （150， 200， 250， 300 mg⋅L-1） demonstrated the increase in inhibition rate of cell proliferation （P<0.01） and cell clone inhibition rate， and decrease in cell clone formation rate （P<0.01）. In comparison with the blank group， STA groups showed typical characteristics of apoptosis， such as chromatin condensation and enhanced fluorescence reaction. The apoptosis rate of STA groups was significantly higher than that of the blank group （P<0.01）. Compared with the blank group， STA （150， 200， 250， 300 mg⋅L-1） significantly up-regulated the protein expression of Caspase-3 and Bax （P<0.05， P<0.01） and down-regulated the expression of Bcl-2 protein （P<0.01）. Compared with the blank group， STA had no significant influence on the total protein expression of PI3K， Akt， JNK， and p38 MAPK. However， STA （150， 200， 250， 300 mg⋅L-1） significantly decreased the levels of p-PI3K and p-Akt （P<0.05， P<0.01） and increased the level of p-p38 MAPK （P<0.05， P<0.01）. Compared with the blank group， STA （200， 250， 300 mg⋅L-1） significantly raised the level of p-JNK （P<0.05， P<0.01）. ConclusionSTA can inhibit the proliferation and induce the apoptosis of A549 cells by inhibiting PI3K/Akt signaling pathway and activating JNK/p38 MAPK signaling pathway.

Exploring the mechanism of elemene synergistic bortezomib against multiple myeloma by ROS-NF-κB-p38MAPK pathway / 中国临床药理学与治疗学

AIM: To investigate the mechanism of elemene synergistic bortezomib against multiple myeloma based on ROS-NF-κB-p38MAPK signaling pathway. METHODS: CCK-8 assay was used to detect cell activity. Nude mice were randomly divided into control group, bortezomib (BTZ) group, elemene (ELE) group and combination group. Each group was treated with BTZ, ELE and BTZ combined with ELE, respectively. Tunel staining was performed to observe the apoptosis of tumor tissues. The expressions of Caspase-3, Bcl-2, NF-κB and p38 MAPK were detected by Western Blot. Cell cycle, apoptosis and reactive oxygen species (ROS) expression were detected by flow cytometry using human myeloma U266 cells. RESULTS: When 4.0 μmol/L ELE combined with 50 nmol/L BTZ treated U266, the cell activity was significantly reduced compared with that of NC, BTZ and ELE groups (P< 0.05). The tumor volume of nude mice in BTZ group, ELE group and combined group was significantly reduced compared with the control group (P <0.05), and the combined group was the smallest. Tunel staining results showed that the apoptosis level in the control group was lower than that in the BTZ group, ELE group and the combined group (P<0.05), and the combined group had the lowest apoptosis level. Compared with the control group, the expressions of Caspase-3 and p38 MAPK in BTZ group, ELE group and combination group were significantly increased, while the expression of Bcl-2 was significantly decreased. The apoptosis level and expression of ROS in BTZ group, ELE group and the combined group was significantly increased compared with the control group (P<0.05). CONCLUSION: ELE can enhance the role of BTZ in promoting apoptosis of myeloma cells, which may be achieved by regulating ROS/NF-κB/p38 MAPK signaling pathway to enhance the level of apoptosis of tumor cells to achieve anti-tumor effect.

Role of FoxO1/p38MAPK signaling pathway in acute lung injury induced by LPS / 中国药理学通报

Aim To investigate the effect of forkhead transcription factors of O classl (FoxO1) on lipopolysaccharide (LPS) -induced acute lung injury and its regulatory mechanism. Methods The model of acute lung injury (ALI) was simulated by LPS. HE staining was used to observe the pathological changes of lung tissues. The contents of tumor necrosis factor a (TNF-a) and interleukin-6 (IL-6) in lung tissues were determined by ELISA. The expression of FoxOl in mouse lung tissues was observed by immunohistochemical staining. The phosphorylation levels of FoxOl, DNA methyltransferase and p38 MAPK were detected by Western blot. The mRNA levels of FoxOl, IL-6, TNF-a and DNA methyltransferase were detected by qRT-PCR. DNA methylation in FoxOl promoter region in lung tissues was detected by nested methylation specific PCR (nMS-PCR). Pulmonary vascular endothelial cells (PVECs) were cultured and transfected with FoxOl siRNA, and the phosphorylation of p38 MAPK was detected by Western blot. The correlation between FoxOl methylation level and inflammatory factors was analyzed by Pearson method. Results Compared with control group, alveolar inflammatory cells increased significantly in LPS group, and pulmonary edema and hyperemia were obvious. TNF-α and IL-6 levels increased by 52. 2% and 150. 4% (P < 0. 05), respectively. The phosphorylation level of p38 MAPK and FoxOl expression increased by 134. 1% and 61. 8% (P < 0. 05), respectively, while the DNA methylation level of Fox0l promoter region decreased by 17. 2% (P < 0. 05). After transfection of FoxOl siRNA in vitro, the phosphorylation level of p38 decreased. Pearson analysis showed that FoxOl methylation level was negatively correlated with inflammatory factors. Conclusion The regulation of FoxOl/p3 8 MAPK signaling pathway by hypomethylation of FoxOl promoter is an important mechanism of LPS-induced acute lung injury.

Dapagliflozin regulates myocardial SK2 protein expression in diabetic rats / 中国药理学通报

Aim To investigate the effect of dapagliflozin on the small conductance calcium-activated potassium channel 2 (SK2 channel) protein in the myocardium of diabetic rats and its possible mechanism of action. Methods In vivo: type 2 diabetes model was established by high-glucose and high-fat diet combined with intraperitoneal injection of low-dose streptozotocin (35 mg

Analysis of the effect of midazolam on pain in a rat model of lumbar disc herniation based on the p38 MAPK signaling pathway / 中国骨伤

OBJECTIVE@#To investigate the effect of midazolam on pain in lumbar disc herniation model rats based on p38 MAPK signaling pathway.@*METHODS@#Fifty SPF-grade Sprague-Dawley healthy rats, half male and half female, were selected and randomly divided into normal group, model group, and low-dose, medium-dose, high-dose groups. Model group and low-dose, medium-dose, high-dose groups were initially modeled for lumbar disc herniation. Intraperitoneal injection of saline was performed in rats of normal and model groups; and in the low-dose, medium-dose, and high-dose groups, intraperitoneal injection of midazolam was performed with doses of 30, 60, and 90 mg/kg, respectively. Interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), 5-hydroxytryptamine (5-HT), β-endorphin (β-EP), substance P (SP), neuropeptide Y (NPY) were detected in the serum of rats by enzyme-linked immunoassay. The expression of p38 MAPK and matrix metalloproteinase-3(MMP-3) protein were detected by Western blot in the tissues of rats of each group.@*RESULTS@#The levels of TNF-α, IL-1β and β-EP were higher and the level of 5-HT was lower in the model group than in the normal group(P<0.05);the levels of TNF-α, IL-1β and β-EP were lower and the level of 5-HT was higher in the low-dose, medium-dose and high-dose groups than in the model group(P<0.05). The levels of SP and NPY increased in the model group compared with the normal group (P<0.05) and the levels of SP and NPY decreased in the low-dose, medium-dose and high-dose groups compared with the model group (P<0.05). The expression of p38 MAPK and MMP-3 increased in the model group compared with the normal group (P<0.05); the expression of p38 MAPK and MMP-3 decreased in the low-dose, medium-dose and high-dose compared with the model group(P<0.05).@*CONCLUSION@#Midazolam may ameliorate the immune inflammatory response in rats with a model of lumbar disc herniation, possibly regulated through the p38MAPK signaling pathway.

Inhibitory effect of hederagenin on the proliferation of bladder cancer cells in vitro and in nude mice through the JNK/p38 MAPK signaling pathway / 中华内分泌外科杂志

Objective:To explore the effect of hederagenin (HE) on the proliferation of bladder cancer cell T24 in vitro and in nude mice.Methods:Human bladder cancer cells T24 were divided into control group and experimental group. The experimental group was cultured with DMEM medium containing 25μg/mL hederogenin, and CCK8 was used to detect cell proliferation ability. Nude mice were divided into a control group and an experimental group and injected with T24 cells. The cells of the experimental group were injected with ivy sapogenin at 30 mg/kg every other day. The protein of T24 cells and tumor mass was extracted to detect the expression of p-JNK/JNK and p-p38/p38.Results:After the bladder cancer cells T24 were treated with hederagenin, the CCK8 results showed that the cell proliferation ability of the experimental group was significantly decreased ( P<0.05) . The expression levels of p-JNK in experimental group and control group were 0.21±0.06 and 0.89±0.15, respectively, and the expression levels of p-p38 were 0.38±0.09 and 1.44±0.26, respectively. The expressions of p-JNK and p-p38 were up-regulated (all P<0.05) . In vivo, it was found that after treatment with ivisaponin, the volume of tumor mass were 1192.07±250.92μm 3 in the subcutaneous tumor experimental group and 2280.50±600.1μm 3 in the control group, and the mass were 0.65±0.29g and 1.62±0.38g, respectively. The mass and volume of the experimental group were decreased (all P<0.05) . We extracted mass proteins, and western blotting results showed that the expression levels of p-JNK in the experimental group and control group were 0.38±0.08 and 1.03±0.19, respectively, and the expression levels of p-p38 were 0.71±0.12 and 1.36±0.25, respectively. The expressions of p-JNK and p-p38 were up-regulated (all P<0.05) . Conclusion:Hederagenin inhibits the proliferation of bladder cancer in vitro and in nude mice through the JNK/p38 MAPK signaling pathway.

Effect of formyl peptide receptor 2 in recurrent spontaneous abortion through p38 MAPK pathway / 解剖学报

Objective To explore the express of formyl peptide receptor 2(FPR2) in villi of recurrent spontaneous abortion ( RSA) , the effect on proliferation, migration and invasion of trophoblast, and the mechanism to clarify the effect of FPR2 on trophoblast function and explore its role and mechanism in recurrent spontaneous abortion. Methods Clinical villus specimens of 30 nonnal and 30 RSA patients were collected. Immunohistochemical staining, Real-time PCR and Western blotting were used to detect the location and expression of FPR2 in villi of patients with RSA and nonnal pregnant women. CRISPR/Cas-9 technique was used to knock down FPR2 in HTR-8/SVneo cells, CCK-8 assay, wound healing and Transwell assays were used to detennine the ability of cell viability, migration and invasion. Immunofluorescent staining and Western blotting were used to analyze the changes of phosphorylated p38 MAPK ( p-p38 MAPK)/p38 MAPK protein expression after applying with p38 MAPK inhibitor SB203580 alone or in combination. Results The expression of FPR2 in villi of patients with RSA increased. FPR2 knock-down improved the biological functions of HTR-8/Svneo cells such as proliferation, migration and invasion significantly. The expression of p-p38 MAPK was up-regulated significantly by FPR2 knock-down, and the ability enhancement of migration and invasion of trophoblasts was reversed partially by SB203580 which inhibits p38 MAPK pathway. FPR2 knock-down caused the change of p38 MAPK signaling pathway related to proteins. Conclusion FPR2 is highly expressed in trophoblasts of RSA patients, and inhibits the migration and invasion of trophoblasts through p38 MAPK signaling pathway, which ma)' play an important role in RSA.

Berberine affects activity of HSV-1 virus infected HEp-2 cells and its molecular mechanism / 中国药理学通报

Aim To discuss the effect of berberine ( BE) on the activity of HSV-1 virus infected HEp-2 cells and its related molecular mechanisms.Methods Hie infected cell model was constructed and divided into control group, infection group, low concentration group ( 5 (xmol • L 1 -BE) , medium concentration group ( 10 (xmol • L '-BE) and high concentration group ( 15 (xmol • L '-BE) ) , and then incubated for 24 hours.qRT-PCR was used to determine HSV-1 infection-related genes ( gD, ICP-4, ICP-8, ICP-27 ) and mRNA expression levels of LncBNA NRAV, miR- 299-3p, RAB5C.CCK-8 method and flow cytometry were applied to detect cell viability and apoptotic rate.The expression levels of PI3K/AKT signaling pathway and JNK/p38 MAPK signaling pathway related protein were analysed by WB.Results It was found that BE j j reduced the mRNA expression of gD, ICP-4, ICP-8, anrl ICP-27, improved cell viability, and inhibited eell apoptosis.BE promoted the expression of miR-299-3p by inhibiting LncRNA NRAV and RAB5C.BE inhibited the protein expression levels of PBK/AKT signaling pathway and JNK/p38 MAPK signaling pathway proteins PI3K, AKT, JNK, and P38.Conclusions The mechanism that BE enhances the activity of HEp-2 cells after HSV-1 infection and suppresses its apoptosis may be related to LncRNA NRAV and RAB5C targeting competitive binding to miH-299-3p, inhibiting the activation of PBK/AKT signaling pathway and JNK/ p38 MAPK signaling pathway.

Effect of Combined Therapy of Lung and Intestine on VIP and p38 MAPK Signaling Pathways in Bronchial Asthma Mice / 中国实验方剂学杂志

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.

Periostin inhibits hypoxia-induced oxidative stress and apoptosis in human periodontal ligament fibroblasts p38 MAPK signaling pathway / 南方医科大学学报

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.

Effects of p38 MAPK signal pathway inhibition on upper lip scar hyperplasia at different time in rabbits / 中华医学美学美容杂志

Objective To investigate the effect of p38MAPK gene silencing recombinant adeno-virus on the expression of target gene in different time and to detect the effect of p 38MAPK signal pathway on the upper lip scar hyperplasia at different time to determine the optimal scar treatment time .Methods The adenovirus vector was injected into the scar tissue in 0 week ,1 week and 2 week after cheiloplasty in rabbit .The specimens were harvested in 3 week postoperatively .Four methods in-cluding Sirius red staining ,immunohistochemical staining (IHC) ,Western blotting (WB) ,real-time PC (RT-PCR) were used to quantitatively and quantitatively detect the relative expression levels of p38MAPK and scar-related factors (col Ⅰ ,col Ⅲ ,MM P1 ,TIMP1) .Results Sirius red staining and immunohistochemical staining showed that in 1st week the expression of col Ⅲ and MMP1 in scar tis-sue was significantly higher than that in 0 week and 2 week after operation and the expression of col Ⅰand TIMP1 was significantly less than that in 0 week and 2 week after operation .The results of WB and RT-PCR were consistent with that of IHC .Conclusions After injection into the upper lip scar tis-sue with adenovirus in 1 week ,the degree of scar hyperplasia is the least .

Effects of brd4 gene on apoptosis and p38mapk signaling pathway of cardiomyocytes induced by tgf-pi / 吉林大学学报(医学版)

Objective: To investigate the effect of bromodomain-containing protein 4 (BRD4) gene on the apoptosis of cardiomy

Effect of inhibition of immunosuppressive factor COX-2 on apoptosis of retinal ganglion cells and regulation of p38MAPK signal pathway / 中国免疫学杂志

Objective: To investigate the effect of COX-2 inhibitor bevacizumab on the activity and apoptosis of retinal ganglion RGC-5 cells induced by H2O2 and the regulation of p38 MAPK signaling pathway. Methods: Retinal ganglion RGC-5 cells was stimulated using H2O2 ( 200, 300, 400, 600, 800 μmol/L) to establish a H2O2 damage model, H2O2 concentration was selected based on half inhibitory concentration, COX-2 inhibitor bevacizumab treated RGC-5 cell induced by H2O2 for 7 h, SB203580 as a p38 MAPK signaling pathway inhibitor, cell viability and apoptosis rate were detected by MTT method and flow cytometry, respectively; the expression of PCNA, p53, p38 and p-p38 protein were detected by Western blot. Results: Different concentrations of H2O2 could inhibit the viability of RGC-5 cells, and the cell viability decreased with the increase of H2O2 concentration, because 400 μmol/L H2O2 inhibited half of the cell viability, it was selected as an object of study. Compared with the control group, the cell viability and the expression of PCNA were decreased significantly in H2O2 group, the apoptosis rate and the expression of p53 and p-p38 protein was increased significantly; compared with H2O2 group, the cell viability and PCNA expression were increased significantly in the H2O2+bevacizumab group, the apoptosis rate and the expression of p53 and p-p38 protein were decreased significantly ( P ＜ 0. 05); compared with H2O2+ bevacizumab group, the cell viability and PCNA expression were increased significantly in H2O2+bevacizumab+SB203580 group, the apoptosis rate and the expression of p53 and p-p38 protein were decreased significantly ( P＜0. 05). Conclusion: Inhibition of immunosuppressive factor COX-2 expression can improve the activity of retinal ganglion cells and inhibit apoptosis by regulating the p38 MAPK signaling pathway.

Effect of osthole on p38 MAPK signaling related pathway of rat model with inflammatory radicular pain induce by nucleus pulposus in the spinal horn / 中国药理学通报

Aim To observe the effect of epidurally application of osthole on the model of nucleus pulposusinduced inflammatory radicular pain and the expression of p38 MAPK signaling related pathway in the spinal dorsal horn of rats.Methods The model of radicular pain was generated by putting nucleus pulposus to the L5 dorsal root ganglion (DRG).50％ MWT was measured using Von Frey filaments to calculate mechanical pain threshold before and after operation.50 μL of 20 g · L-1 osthole was administered epidurally in group Ost and 50 μL of 100 mL · L-1 DMSO in group DMSO at postoperative day (POD).The expression of phosphorylated p38 (p-p38),IL-18 and IL-18R in the lumbar spinal dorsal horn was detected by Western blot.IL-18 mRNA was assessed by real-time PCR.Results The mechanical pain threshold significantly decreased after operation (P ＜ 0.05),while the expression of protein p-p38 MAPK,IL-18,IL-18R and IL-18 mRNA was significantly different.Compared with DMSO group,50％ MWT was significantly increased and accompanied with the decrease of protein p-p38,IL-18,IL-lgR and IL-18 mRNA in Ost group after drug administration (P ＜ 0.05).The correlation analysis between protein concentration of p38 MAPK and IL-18 mRNA showed that the Spearman correlation coefficient was 0.9 (P ＜ 0.05).Conclusion p-p38 and IL-18 of spinal dorsal horn participate in the rat model with inflammatory radicular pain induced by nucleus pulposus,and IL-18R plays a role in maintenance of the pain.Osthole administered epidurally in the early stage of pain could alleviate the pain for a long time,which may be related with inhibiting p38 MAPK signaling related pathways.

Advanced oxidation protein products modulate autophagy of renal tubule epithelial cells / 重庆医学

Objective To investigate the effect of advanced oxidation protein products (AOPPs) on human renal tubular epithelial cells(HK-2) autophagy.Methods HK-2 cells were stimulated with AOPPs.RT-qPCR and Western blot were used to determine the expression of autophagy related protein LC3-Ⅱ/LC3-Ⅰ,Beclin1 and p62;Western blot was utilized to examine the activation of p38 MAPK pathway.Then p38 MAPK inhibitor (SB203580) was added and co-processed with AOPPs.The change of autophagy was observed Also,autophagy inducer rapamycin was added and co-processed with AOPPs.RT-qPCR and Western blot were used to detect the expression of cell cycle inhibitory protein p27.The cell total protein level was detected by the bicinchoninic acid (BCA) method.The hypertrophy change was observed.Results AOPPs down-regulated the expression of LC3-Ⅱ/LC3-Ⅰ and Beclin1,up-regulated expression of p62 and activated p38 MAPK pathway;in comparison with the AOPPs alone treatment group,the expression of LC3-Ⅱ/LC3-Ⅰ and Beclin in the SB203580 co-processing group was increased,while p62 was decreased;the p27 expression and cells total protein in the sirolimus co-processing group were down-regulated.Conclusion AOPPs inhibits the autophagy of HK-2 cells by activating p38 MAPK pathway and the autophagy inhibition participates in HK-2 cell hypertrophy.

Effect of leech on VSMCs in early atherosclerosis rats via p38MAPK signaling pathway / 中国中药杂志

To explore the effect of leech on proliferation and apoptosis of vascular smooth muscle cells(VSMCs) in early atherosclerosis rats via p38MAPK signaling pathway and investigate its possible mechanism. Biochemical analyzer was used to examine the regulation of leech on levels of triglycerides(TG), total cholesterol(TC), low-density lipoprotein(LDL-C), and high-density lipoprotein(HDL-C) in blood lipid of rats. The expression of transforming growth factor-beta 1(TGF-β1) in serum was detected by ELISA. Immunological histological chemistry (IHC) was taken to measure the expression levels of proliferating cell nucleus antigen(PCNA) and cell apoptosis proteinase-3(Caspase-3), while the protein expression levels of MKK3, p38 and C-myc were detected by Western blot. In addition, hematoxylin and eosin(HE) staining was used to observe the morphological change of thoracic aortas. The results showed that leech decreased the levels of TC, LDL-C obviously and increased HDL-C, suppressed the expression levels of TGF-β1 and PCNA, up-regulated Caspase-3, down-regulated the expression levels of MKK3, p38, and C-myc protein. HE staining indicated that it could inhibit intimal thickening and reduce plaque formation. The above results indicated that leech may affect the protein expression of the p38MAPK signaling pathway to inhibit proliferation and promote the apoptosis of VSMCs via reducing blood lipid levels and suppressing TGF-β1, aiming at inhibiting intimal thickening and reducing plaque formation, tand then slowing down the process of early atherosclerosis.

Research progress of bone morphogenetic proteins-9 induced periodontal ligament stem cells osteogenesis differentiation and it′s signal pathway / 医学研究生学报

Bone morphogenetic proteins (BMPs) are members of the transforming growth factor-β(TGF-β) superfamily and play critical roles in skeletal development, bone formation and stem cell differentiation.BMP9 is one of the most osteogenic BMPs, promoting osteogenesis differentiation of periodontal ligament stem cells ( PDLSCs) both in vitro and in vivo.Recently, signal transduc-tion studies have revealed that BMP-Receptor-Smads and BMP-Receptor-Mitogen activated protein kinase ( MAPK) play vital roles in BMP9 which promote PDLSCs osteogenesis differentiation.Several studies revealed that transcription factors closely associated with os-teogenesis differentiation are found in the downstream of the Smads and MAPK pathways.This review aims to summarize our current knowledge of BMP9-mediated osteogenesis by presenting recently completed works which may help us to further elucidate these path-ways.