Mechanism of Qiyu Sanlong Prescription in Inhibiting "Addiction" of Lung Cancer A549 Cells to miRNA21 / 中国实验方剂学杂志

Objective:To investigate the molecular mechanism of Qiyu Sanlong prescription （QYSL） in inhibiting the "addiction" of lung cancer A549 cells to miRNA21. Method:The human lung cancer A549 cells were routinely passaged and divided into the blank group， blank serum group， QYSL-containing serum group， and siRNA group. The prepared QYSL-containing serum was used for intervention， with the optimal concentration and action time determined in previous studies. The protein and mRNA expression levels of miRNA21 and related molecules in its target phosphatase and tensin homolog deleted on chromosome 10 （PTEN）/phosphatidylinositol 3-kinase （PI3K） signaling pathway were detected by real-time polymerase chain reaction （Real-time PCR） and Western blot assay. Result:The comparison with the blank serum group revealed that the mRNA expression levels of miRNA21 in the QYSL-containing serum group and the siRNA group were decreased， while the PTEN mRNA expression in the QYSL-containing serum group was increased， showing significant differences （<italic>P</italic><0.01）. Compared with the blank serum， the QYSL-containing serum and siRNA significantly down-regulated PI3K and mammalian target of rapamycin （mTOR） mRNA expression （<italic>P</italic><0.01）， whereas the QYSL-containing serum did not change the mRNA expression of protein kinase B （Akt）. The protein expression levels of PTEN in the QYSL-containing serum group and the siRNA group were obviously elevated in contrast to that in the blank serum group （<italic>P</italic><0.05）. Meanwhile， the protein expression levels of phosphorylated Akt （p-Akt） and phosphorylated mTOR （p-mTOR） evidently declined in the QYSL-containing serum group （<italic>P</italic><0.05）， but there was no significant reduction in total Akt and mTOR protein expression. The PI3K protein expression was slightly down-regulated， with no statistical significance. Conclusion:QYSL inhibits the transcription of miRNA21， increases the expression of PTEN， and reduces the expression of key molecules in PI3K/Akt/mTOR signaling pathway， thus mildly inhibiting the "addiction" of lung cancer cells to oncogenes and blocking their proliferation.

Effect of Erxian Decoction on proteomics and PI3K signaling pathway of osteoblasts stimulated by H2O2 / 中国中药杂志

The present study aimed to explore the effect of Erxian Decoction on proteomics of osteoblasts stimulated by hydrogen peroxide(H_2O_2) and its protective mechanism with the H_2O_2-induced cell model of oxidative stress. The primary osteoblasts were cultured from the skulls of newborn rats(within 24 hours) and divided into a control group, a model group, a Fosamax group, and an Erxian Decoction group. Blank serum was added in the control group and model group, and the drug-containing serum was added correspondingly to the remaining two groups. After 45 hours, H_2O_(2 )stimulation was conducted for three hours except for the control group, followed by protein extraction. Nano-LC-LTQ-Orbitrap system was used for protein detection, Protein Discovery for protein identification, and SIEVE for quantitative and qualitative analysis. Furthermore, following the blocking of PI3 K signaling pathway by LY294002(10 μmol·L~(-1)), a control group, a model group, an LY294002 group, an Erxian Decoction group, and an Erxian Decoction + LY294002 group were set up to observe the effect of Erxian Decoction on cell proliferation, alkaline phosphatase(ALP) activity, and the relative expression of BMP-2, OPG, p-Akt, p-FoxO1 of osteoblasts stimulated by H_2O_2 under LY294002 intervention. The results revealed that 78 differential proteins were discovered between the Erxian Decoction group and model group, which were involved in the regulation of PI3 K/Akt, glucagon, estrogen, insulin, and other signaling pathways. LY294002 blunted the promoting effect of Erxian Decoction on osteoblast proliferation and significantly down-regulated the expression of OPG and p-FoxO1, whereas its down-regulation on the expression of BMP-2 and p-Akt was not significant. Both LY294002 and Erxian Decoction increased the ALP activity of osteoblasts, which may be related to the cell state and the cell differentiation. The above results suggest that Erxian Decoction can protect osteoblasts stimulated by H_2O_2, with the PI3 K/Akt signaling pathway as one of the internal mechanisms.

Hydrogen sulfide attenuates urosepsis-induced acute kidney injury by blocking TLR4/MyD88/PI3K signaling pathway / 中国病理生理杂志

AIM:To explore the effect of hydrogen sulfide (H2S) on urosepsis-induced acute kidney injury.METHODS:New Zealand white rabbits were randomly divided into control group, sham group, model (sepsis) group, Na HS treatment (Na HS) group, and Na HS combined with TAK-242 (a TLR4 inhibitor) treatment (Na HS+TAK-242) group.After treatment for 72 h, HE staining was used to measure the histopathological changes of rabbit kidney.The levels of blood urea nitrogen (BUN) and serum creatinine (SCr) were detected by automatic biochemical analyzer.The serum levels of neutrophil gelatinase-associated lipocalin (NGAL) , kidney injury molecule 1 (KIM-1) , procalcitonin (PCT) , interleukin-1β (IL-1β) , interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by ELISA.The TLR4/MyD88/PI3K signaling pathway-related proteins in the kidney were determined by Western blot.RESULTS:Compared with control group, obvious damage was observed in the kidneys of septic rabbits, but the kidneys were markedly improved by treatment with Na HS.The levels of BUN, SCr, NGAL, KIM-1, PCT, IL-1β, IL-6 and TNF-αin the septic rabbits were higher than those in control group, and decreased significantly in Na HS group and Na HS+TAK-242 group.The protein levels of TLR4, MyD88, p-PI3K and p-Akt in septic rabbit kidneys were higher than those in control group.However, Na HS or Na HS+TAK-242 inhibited the activation of TLR4/MyD88/PI3K signaling pathway in the kidneys of septic rabbits.CONCLUSION:H2S play a protective effect on the rabbits with urosepsis-induced acute kidney injury by blocking TLR4/MyD88/PI3K signaling pathway to inhibit inflammatory response.

Porhyromonas endodontalis liopolysaccharide regulates interleukin-23 expression in mouse osteoblasts / 实用口腔医学杂志

Objective:To investigate the effects of Porhyromonas endodontalis(P.e)liopolysaccharide(LPS)on the expression of IL-23mRNA and protein in mouse osteoblasts MC3T3-E1 and the role of phosphatidylinositol 3-Kinases (PI3K)signaling pathway in this process.Methods:MC3T3-E1 cells were treated with different concentrations of P.e LPS for different hours,or pretreated with LY294002,a special PI3K inhibitor.The IL-23 mRNA and protein expression levels were detected by real-time PCR and Western blot respectively.Results:The level of IL-23 mRNA increased in MC3T3-E1 cells with the increase of concentration and the treatment time of P.e LPS(P <0.05).The IL-23 protein expression was increased by P.e LPS in a time dependent manner(P <0.05).The mRNA and protein of IL-23 decreased(P <0.05)after pretreatment with LY294002.Conclusion:P.e LPS can induce the expression of IL-23 mRNA and protein in MC3T3-E1 cells,and the PI3K signaling pathway may play a part in this process.

The effect and mechanism of epithelial growth factor receptor (EGFR)in inducing vasculogenic mimicry formation / 西安交通大学学报(医学版)

Objective To investigate the effect and mechanism of EGFR in inducing vasculogenic mimicry (VM)formation.Methods Immunohistochemistry was used to detect the expression of EGFR protein and CD34-PAS double staining was used to detect the VM in gliomas paraffin blocks.The migration and VM formation of U87 cell in hypoxic and normoxic groups were detected by transwell and three-dimensional culture.Then we used Western blot to detect the expressions of EGFR,AKT and PI3K protein.Results The positive rate of EGFR expression in VM positive patients was 94.4%,which was significantly higher than that in VM negative patients (71.8%).In hypoxic group,the abilities of migration and VM formation were significantly higher those in normoxic group.The expressions of EGFR,AKT and PI3K were higher in hypoxic group.Conclusion VM formation can be induced by the activation of EGFR/AKT/PI3K signaling pathway.EGFR is very important for VM formation.

HIV-1 Nef promotes KSHV vIL-6-induced angiogenesis by regulating PTEN/PI3K signaling pathway / 中华微生物学和免疫学杂志

Objective To investigate whether HIV-1 Nef could promote the angiogenesis and tu-morigenesis induced by KSHV vIL-6 through regulating PTEN/PI3K signaling pathway .Methods Lipo-some transfection was used to transfect cDNA of pPTEN , dominant-negative ( DN) construct of PI3K and control vector into endothelial cells , which stably express KSHV vIL-6 and HIV-1 Nef.Microtubule forma-tion assay and chicken chorioallantoic membrane ( CAM) assay were used to evaluate microtubule formation and angiogenesis , respectively .Expressions of PTEN and PI 3 K were measured by Western blot .Results Both overexpression of PTEN and inhibited expression of PI 3K suppressed the vIL-6-induced microtubule for-mation and angiogenesis in CAM mediated by Nef .Conclusion HIV-1 Nef enhances vIL-6-induced angio-genesis and tumorigenesis through regulating PTEN/PI3K signaling pathway .