1.Effects of propofol on GRP78-ATF4-CHOP signaling pathway during apoptosis in gastric cancer cells
Miao ZHANG ; Wei YUE ; Xiaodong GU
Chinese Journal of Anesthesiology 2021;41(2):239-241
Objective:To evaluate the effects of propofol on glucose-regulated protein 78 (GRP78)-activating transcription factor-4 (ATF4)-CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP) signaling pathway during apoptosis in gastric cancer cells.Methods:MGC-803 cells were cultured to the logarithmic phase and divided into 2 groups ( n=15 each) using the random number table method: control group (group C) and propofol group (group P). The cells in group C were cultured under normal conditions of 37 ℃ and 5% CO 2. In group P, propofol (final concentration 5 μg/ml) was added when the cells were attached to the wall and grown to 70%-90% confluence.At 24 h after propofol was added, the apoptosis rate of cells was determined by flow cytometry, the expression of GRP78, ATF4 and CHOP mRNA was detected by quantitative polymerase chain reaction, and the expression of GRP78, ATF4 and CHOP was determined by Wester blot, respectively. Results:Compared with group C, the apoptosis rate was significantly increased, and expression of GRP78, ATF4, CHOP and its mRNA was up-regulated in group P ( P<0.05). Conclusion:The mechanism by which propofol accelerates apoptosis is related to activating GRP78-ATF4-CHOP signaling pathway in gastric cancer cells.
2.Multi-level identification and analysis about infrared spectroscopy of lophatheri herba.
Ying SHAO ; Qi-Nan WU ; Wei GU ; Wei YUE ; Da-Wei WU ; Xiu-He FAN
China Journal of Chinese Materia Medica 2014;39(9):1644-1649
Based on the infrared spectra of Lophatheri Herba and Commelinae Herba, one-dimensional infrared spectra, second derivative spectra and two-dimensional correlated spectra were used to find out the differences between Lophatheri Herba and its imitations, respectively. The common peak ratio and variant peak ratio dual-indexes sequential were calculated and established according to infrared spectra of eleven batches of herbs. Infrared spectral data of Lophatheri Herba cluster analysis was applied to explore the similarity between each sample. The grouping results trend of sequential analysis of dual-indexes and cluster analysis was accordant. The results showed that the differences could be found by multi-level identification, and the source and the quality of the herbs could be effectively distinguished by the two analysis methods. Infrared spectroscopy, used in the present work exhibited some advantages on quick procedures, less sample required, and reliable results, which could provide a new method for the identification of traditional Chinese medicine with the imitations and adulterants, and the control of quality and origin.
Drugs, Chinese Herbal
;
analysis
;
chemistry
;
Plant Leaves
;
chemistry
;
Plants, Medicinal
;
chemistry
;
Reproducibility of Results
;
Spectrophotometry, Infrared
;
methods
;
Spectroscopy, Fourier Transform Infrared
;
methods
3.Effects of procyanidin from vaccinium on the myocardial fibrosis in rats
Hongyan FAN ; Yanchun WANG ; Kuang REN ; Raosheng GU ; Nan SHEN ; Wei LIU ; Yue ZHONG
Chinese Pharmacological Bulletin 1986;0(05):-
Aim To explore the effect of Procyanidin from Vaccinium(PC) on the myocardial fibrosis and its mechanism.Methods The model of myocardial fibrosis in rats was built by subcutaneous injection of isoprenaline(Iso) for 5 mg?kg-1?d-1 in vivo.Meanwhile,rats were treated with PC 100,200 and 400 mg?kg-1?d-1 by gastrointestinal administration.The contents of hydroxyproline(Hyp),malondialdehyde(MDA) and the activity of superoxide dismutase(SOD) in left ventricle were assayed with spectrophotometry respectively.Changes of myocardial ultrastructure were observed by electronic microscope.The cultured neonatal rat CFb was isolated by trypsin digestion method.The proliferation of CFb was induced by Angiotensin Ⅱ and assessed by thiazolyl blue(MTT) assay.PC was added in different dosage(25,50 and 100 mmol?L-1).Ⅰ,Ⅲ collagen quantification and TGF-?1 levels were determined by the ELISA.Results In vivo,compared with model group,the contents of hydroxyproline and malondialdehyde of left ventricle were significantly reduced and the activity of superoxide dismutasein left ventricle was markedly enhanced in PC three groups(P
4.An overview of mechanism research on Helicobacter pylori pathogenic factors
Yue BI ; Wei LUO ; Hai-Ying GU
Basic & Clinical Medicine 2018;38(5):713-716
Helicobacter pylori(Hp)is considered to be closely associated with gastritis,peptic ulcer,gastric cancer and other gastrointestinal diseases,but the detail of the pathogenesis is still unclear.The past researches often fo-cused on virulence factors such as CagA(cytotoxin-associated gene/protein A),VacA(vacuolating cytotoxin A), Urease and DupA(duodenal ulcer-promoting gene/protein A),but these are not enough to demonstrate the patho-genic mechanism.Recent studies have not only revealed some new progresses of these past virulence factors but also found that Hp flagllum,adhesion protein and hemolysin play important roles in the pathogenic mechanism.
5.Congenital salivary gland anlage tumor: report of a case.
Long LIN ; Hong-Feng TANG ; Yue-Feng SUN ; Wei-Zhong GU ; Hua-Ying YE
Chinese Journal of Pathology 2009;38(10):711-712
Actins
;
metabolism
;
Adenoma, Pleomorphic
;
congenital
;
metabolism
;
pathology
;
surgery
;
Diagnosis, Differential
;
Fibrosarcoma
;
metabolism
;
pathology
;
Humans
;
Infant
;
Male
;
Nasopharyngeal Neoplasms
;
congenital
;
metabolism
;
pathology
;
surgery
;
Rhabdomyosarcoma
;
metabolism
;
pathology
;
Salivary Gland Neoplasms
;
congenital
;
metabolism
;
pathology
;
surgery
;
Vimentin
;
metabolism
6.Molecular mechanism of emodin on inhibiting autophagy induced by HBSS in renal tubular cells.
Hao HU ; Wei SUN ; Liu-bao GU ; Yue TU ; Hong LIU
China Journal of Chinese Materia Medica 2015;40(10):1965-1970
OBJECTIVETo explore the regulative effects and possible mechanisms of emodin on autophagy induced by starvation in rat's renal tubular epithelial cells (NRK-52E).
METHODFirstly, Hank's balanced salt solution (HBSS) was used to induce starvation and the protein expression of microtubule-associated protein 1 light chain 3 (LC3) I/II, an autophagic marker of mammalian congener, was detected by Western blot with or without the treatment of emodin. Secondly, the changes of red fluorescent protein-microtubule associated protein light chain3 (RFP-LC3) fluorescent particles, treated by HBSS (1 mL) and bafilomycin A1 (10 nmol x L(-1)) with or without emodin, were observed through fluorescence microscopy in NRK-52E cells transient transfected by RFP-LC3 plasmid. With the intervention of mammalian target of rapamycin mTOR inhibitor rapamycin (100 nmol x L(-1)) , the effect of blocking mTOR signaling pathway on autophagic inhibition of emodin was observed. Finally, the effect of mTOR signaling pathway on autophagic inhibition of emodin was further evaluated through the over-expression of endogenous mTOR inhibitory protein DEP domain-containing mTOR-interacting protein-(DEPTOR).
RESULTHBSS hunger could induce high protein expression of LC3 II in NRK-52E cells, and the intervention of emodin could reverse the unregulated protein expression of LC3 II induced by HBSS. The number of RFP-LC3 fluorescent particles was increased after the co-treatment of HBSS and bafilomycin A1, and this increase was inhibited by emodin. After the co-treatment of rapamycin, emodin and HBSS, the LC3 II protein expression restored in NRK-52E cells, compared with the treatment of HBSS. Over-expression of DEPTOR could also block the inhibitive effect of emodin on LC3 II protein expression.
CONCLUSIONEmodin could inhibit HBSS-induced LC3 II protein expression and the activation of autophagy in NRK-52E cells, and the effect of blocking autophagy may be mediated through mTOR signaling pathway.
Animals ; Autophagy ; drug effects ; Cell Line ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Emodin ; pharmacology ; Isotonic Solutions ; adverse effects ; Kidney Tubules ; cytology ; drug effects ; metabolism ; Microtubule-Associated Proteins ; genetics ; metabolism ; Rats ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; genetics ; metabolism
7.Effects of movement on hippocampus β-amyloid protein and amyloid precursor protein in senescence-accelerated/prone mice
Bingjie WU ; Jianyong JIANG ; Yonghong SUN ; Wei YUE ; Yumiao ZHANG ; Min LIU ; Ping GU ; Mingwei WANG
Chinese Journal of Physical Medicine and Rehabilitation 2011;33(1):2-5
Objective To explore the effects of movement on hippocampal β-amyloid protein ( Aβ ) and amyloid precursor protein (APP) in senescence-accelerated and senescence-prone (SAMP8) mice, and the mechanism by which movement improves learning and memory in mice with a model of Alzheimer's disease. Methods Forty 3-month-old SAMP8 mice were divided randomly into a movement group and a control group. The movement group was trained with a running wheel 10 min daily, 5 days a week in the first month, and 20 min daily in the second month. Morphological changes in the hippocampus were observed under the microscope after HE staining. The expression of Aβ in the hippocampus was detected by immumohistochemical methods and APP mRNA expression was detected by RT-PCR two months later. Results HE staining showed neuron degeneration and death, chromatin condensation and vacuolar degeneration in the hippocampus of the 5-mouth-old SAMP8 mice of the control group. The movement group showed less neuron degeneration and death, and the morphology of most cells was normal The expression of Aβ in the hippocampus of the 5-month-old SAMP8 mice in the movement group was significantly lower than that in the control group. APP mRNA expression levels in the movement group were also significantly lower.Conclusions Movement can delay neuron degeneration and down-regulate Aβ and APP mRNA expression levels in the hippocampus of SAMP8 mice. It may be an important mechanism by which movement improves learning and memory in mice with a model of Alzheimer's disease.
8.Mechanisms of cordycepin on improving renal interstitial fibrosis via regulating eIF2α/TGF-β/Smad signaling pathway.
Liu-bao GU ; Rong-wen BIAN ; Yue TU ; Hao HU ; Yi-gang WAN ; Wei SUN
China Journal of Chinese Materia Medica 2014;39(21):4096-4101
OBJECTIVETo investigate the effects and mechanisms of cordycepin,an effective component of cordyceps militaris, on renal interstitial fibrosis (RIF) and its related eIF2α/TGF-β/Smad signaling pathway.
METHODFirstly, 15 C57BL/6 mice were randomly divided into 3 groups,the control group (Group A), the model group (Group B) and the cordycepin-treated group (Group C). After renal interstitial fibrotic model was successfully established by unilateral ureteral obstruction (UUO), the mice in Group C were intraperitoneally administrated with cordycepin(5 mg x kg(-1) d(-1)) and the ones in Group A and B were administrated with physiological saline for 5 days. At the end of the study, the obstructed kidneys were collected and detected for the pathological changes of RIF, and the mRNA expressions of collagen type I (Col I) and α-smooth muscle actin (α-SMA) in the kidney by Northern blot. Secondly, after renal tubular epithelial (NRK-52E) cells cultured in vitro were exposed to transforming growth factor (TGF) -β with or without cordycepin, the mRNA expressions of Col I and collagen type IV( Col IV) by Northern blot, and the protein expressions of eukaryotic initiation factor 2α (eIF2α), phosphorylated eIF2α ( p-eIF2α), Smad2/3 and phosphorylated Smad2/3 (p-Smad2/3) were tested by Western blot.
RESULTIn vivo, cordycepin alleviated RIF in model mice, including improving fibrotic pathological characteristics and mRNA expressions of Col I and α-SMA. In vitro, cordycepin induced the high expression of p-elF2α, and inhibited the expressions of p-Smad2/3, Col I and Col IV induced by TGF-β in NRK-52E cells.
CONCLUSIONCordycepin attenuates RIF in vivo and in vitro, probably by inducing the phosphorylation of eIF2α, suppressing the expression of p-Smad2/3, a key signaling molecule in TGF-β/Smad signaling pathway, and reducing the expressions of collagens and α-SMA in the kidney.
Actins ; analysis ; Animals ; Deoxyadenosines ; pharmacology ; Fibrosis ; Kidney ; drug effects ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Phosphorylation ; Protein-Serine-Threonine Kinases ; physiology ; Signal Transduction ; drug effects ; Smad Proteins ; physiology ; Transforming Growth Factor beta ; antagonists & inhibitors ; physiology
9.Molecular mechanism of rhein on inhibiting autophagic protein expression in renal tubular epithelial cells via regulating mTOR signaling pathway activation.
Yue TU ; Wei SUN ; Liu-bao GU ; Yi-Gang WAN ; Hao HU ; Hong LIU
China Journal of Chinese Materia Medica 2014;39(21):4090-4095
OBJECTIVETo explore the effects and molecular mechanisms of rhein on reducing starvation-induced autophagic protein expression in renal tubular epithelial ( NRK-52E) cells.
METHODHank's balanced salt solution (HBSS) was used to induce NRK-52E cells to be in the state of starvation. After the intervention of HBSS for 0, 0.5,1, 2 and 6 hours, firstly, the protein expression of microtubule-associated protein 1 light chain 3(LC3 I/II), which is a key protein in autophagy, was detected. Secondly, the protein expressions of mammalian target of rapamycin (mTOR) and phosphorylated-mTOR Ser2448 (p-mTOR S2448) were examined. And then, after the co-treatment of rhein (5 mg x L(-1)) and HBSS (1 mL) without or with mTOR inhibitor, rapamycin (100 nmol x L(-1)), the protein expressions of LC3 I/II, mTOR and p-mTOR S2448 were tested, respectively.
RESULTHBSS could induce the up-regulation of LC3 II and the down-regulation of p-mTOR S2448 at protein expression level in NRK-52E cells. The co-treatment of rhein and HBSS could reversely regulate the protein expressions of LC3 II and p-mTOR S2448 in NRK-52E cells significantly. The co-treatment of rapamycin, rhein and HBSS could recover the level of LC3 II protein expression in HBSS-intervened NRK-52E cells.
CONCLUSIONHBSS induces autophagy in renal tubular epithelial cells by inhibiting mTOR signaling pathway activation. Rhein reduces the autophagic protein expression in renal tubular epithelial cells through regulating mTOR signaling pathway activation, which is the possible effects and molecular mechanisms.
Animals ; Anthraquinones ; pharmacology ; Autophagy ; drug effects ; Cells, Cultured ; Epithelial Cells ; drug effects ; metabolism ; Isotonic Solutions ; pharmacology ; Kidney Tubules ; drug effects ; metabolism ; Microtubule-Associated Proteins ; genetics ; Rats ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; antagonists & inhibitors ; genetics ; physiology
10.An efficient method which can be used to transfect Tibetan minipig embryonic fibroblasts
Wei LIU ; Yan CHEN ; Min YUE ; Jin YUAN ; Tianwu QIU ; Dong XIAO ; Weiwang GU
Chinese Journal of Comparative Medicine 2015;(3):64-66
Objective To transfect EGFP gene to porcine embryonic fibroblasts ( PEFs) of Tibetan miniature pigs by Lonza Nucleofector II machine and compare the tansfection efficiency between this method and the lipofection method. Method A plasmid carrying green fluorescent protein ( GFP) was transfected into PEFs of Tibetan miniature pigs via the Lonza Nucleofector II machine ( program U020) and by Lipofectamine 2000.Results 5 hours after nucleofection, green fluorescence was observed, indicating 80%transfecting efficiency in the nucleofection group, which is significantly higher than the lipofection group. Conclusion Nucleofector II machine can efficiently transfect PEFs, provides a reliable method for efficiently generate transgenic Tibetan minipigs.