Animals ; Coronary Restenosis ; prevention & control ; Diterpenes ; administration & dosage ; Epoxy Compounds ; Iliac Artery ; pathology ; Male ; Phenanthrenes ; administration & dosage ; Rabbits ; Stents ; Tunica Intima ; pathology

Objective To explore whether extracellular signal-reg-ulated kinase (ERK) signaling pathway is involved in the hypoxia-inducible factor-1α (HIF-1α) expression in the hippocampus of epileptic rats.Methods A total of 208 21-d old SD rats were equally randomized into status epilepticusgroup (SE,n=96),normal control group (NS,n=96) and PD98059 (the ERKsignaling pathway specific inhibitor) treatment group (n=16),respectively; SE rat models of the SE group were induced by intraperitoneal injection of 1％ PTZ (80 mg/kg),and rats of the NS group received injection of normal saline (NS); 0.5,1,1.5,6,12 and 24 h after the inducement,the mRNA and protein expressions of HIF-lα and ERK1/2 in the hippocampus of rats in these two groups were examined by RT-PCR and Westem blotting.For rats in the PD98059 treatment group,PD98059 was intraperitoneally injected 10 minutes before intraperitoneal injection of pentylenetetrazol; 1 h after the inducement,the mRNA expressions of HIF-1α and ERK1/2 in the hippocampus of rats were examined by RT-PCR,while 1.5 h after the inducement,the protein expressions of HIF-1α and ERK1/2 in the hippocampus of rats were examined by Western blotting.The results of the PD98059 treatment group would be compared with those of the SE group.Results Compared with the NS group,the mRNA and protein expressions of HIF-1α and ERK 1/2 in the hippocampus of rats in the SE group after SE increased significantly; the peakexpression time ofERK1/2 mRNA was 1 h after SE (1.112±0.126 h),and the ERK1/2 protein mostly expressed at 1.5 h after SE (1.127±0.155 h).As to HIF-1α mRNA and its protein,the peak expression time was 1.5 h after SE (0.589±0.090 h) and 6 h after SE (0.230±0.052 h),respectively (P＜0.05).Compared with the SE group,the mRNA and protein expressions of HIF-1α and ERK1/2 in the hippocampus of all the rats in the PD98059 treatment group after SE decreased significantly (P＜0.05);positive correlation between HIF-1α and ERK1/2 mRNA in the SE group was noted (r=0.688,P=0.000).Conclusion ERK signaling pathway is activated in the epileptic rats and it participates in the expressionof HIF-1α in the hippocampus.

OBJECTIVETo study on the effect of acteoside on learning and memory of dementia mice.

METHODMice were orally administered with acteoside for 10 days. Scopolamine was used to establish the acquired learning disability in mice. Their learning and memory were detected with a behavioral experiment (step-down test). After the behavior test, corticocerebral and hippocampus tissues of mice were detected with biochemical indexes, including GSH-Px, T-SOD, MDA, TChE and contents of protein in brain tissues.

RESULTMice were administered with acteoside for 10 d in advance to alleviate the acquired learning disability induced by scopolamine. Compared with the model group, acteoside increased the latency period in the step-down test and reduced error times. Besides, acteoside increased the activity of GSH-Px, T-SOD, TChE and protein content in their brain tissues, but decreased MDA content.

CONCLUSIONActeoside can significantly alleviate the acquired learning disability in mice induced by scopolamine. Its mechanism may be related with its effect of inhibiting the generation of free radicals in mice and improving the function of the central cholinergic system.

Animals ; Behavior, Animal ; drug effects ; Brain ; drug effects ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Glucosides ; administration & dosage ; pharmacology ; Glutathione Peroxidase ; metabolism ; Learning ; drug effects ; Male ; Memory Disorders ; chemically induced ; drug therapy ; Mice ; Phenols ; administration & dosage ; pharmacology ; Scopolamine Hydrobromide ; adverse effects ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the effect of acteoside on SK-N-SH nerve cell injury induced by okadaic acid (OA).

METHODSK-N-SH nerve cells were processed with 20 nmol * L OA to establish the Alzheimer's disease (AD) cellular model, and 5, 10, 20 mg . L-1 acteoside was used to antagonize against its effect. Cell morphology was observed under inverted microscope. The cell survival rate was detected with MTT, and the LDH release rate was measured by enzyme label kit. Western blot was applied to determine the expression of phosphorylation tau proteins in nerve cells.

RESULTThe acteoside could significantly improve SK-N-SH cell morphology, enhance the cell survival rate, decrease the cell LDH release rate and the expression of phosphorylated tau proteins at p-Ser 199/202 and p-Ser 404 sites, up-regulated the expression of at non-phosphorylated tau proteins at Ser 202 site and Ser 404 sites.

CONCLUSIONActeoside has significant protective effect on nerve cell injury induced by OA.

Alzheimer Disease ; metabolism ; Cell Line ; Cell Survival ; drug effects ; Glucosides ; pharmacology ; Humans ; Okadaic Acid ; Phenols ; pharmacology ; tau Proteins ; metabolism

OBJECTIVETo detect the expression of phosphorylated-signal transducer and activator of transcription 3 (p-Stat3) and myeloid leukemia-1 (Mcl-1) as well as their correlation, and to investigate the functional role of Stat3 and Mcl-1 in the pathogenesis of esophageal squamous cell carcinoma (ESCC).

METHODSStat3 activity in ESCC cells was inhibited with JAK/Stat3 inhibitors (AG490 or JSI-124). Specific siRNA was used to inhibit the Stat3 expression. Cell apoptosis was detected by flow cytometry. Expression of Mcl-1 protein was determined by Western blotting. Expression of phospho-Stat3 (Tyr705) and myeloid leukemia-1 (Mcl-1) proteins in ESCC tissues was detected by tissue microarray and immunohistochemistry. The relationship between p-Stat3 or Mcl-1 aberrant expression and clinicopatholohical features of ESCC was analyzed. The correlation of their expression was also analyzed.

RESULTSSuppression of the Stat3 signaling activation in ESCC cells led to marked apoptosis, and dramatic reduction of Mcl-1 protein. The positive rate of phospho-Stat3 (Tyr705) expression was 45.0% in 50/111 of the ESCC tissue samples. The lower the degree of tumor differentiation, the higher the positive rate of phospho-Stat3 (Tyr705), showing a significant difference (P = 0.018). The positive rate of Mcl-1 protein expression was 72.1% (80/111), and the lower the degree of tumor differentiation was, the higher there was the positive rate of Mcl-1, with a significant difference (P = 0.026). There was a positive correlation between the expressions of p-Stat3 and Mcl-1 proteins (P = 0.012).

CONCLUSIONSIn a subset of ESCC tissues, p-Stat3 (Tyr705) and Mcl-1 are overexpressed and positively correlated with each other, and both are correlated with tumor differentiation. Persistent activation of Stat3 contributes to apoptotic resistance in ESCC cells, and may be at least partly mediated through upregulation of Mcl-1.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Cell Line, Tumor ; Cell Survival ; drug effects ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Neoplasm Grading ; Neoplasm Staging ; Phosphorylation ; RNA, Small Interfering ; genetics ; STAT3 Transcription Factor ; antagonists & inhibitors ; genetics ; metabolism ; Tyrphostins ; pharmacology

OBJECTIVETo establish a HPLC method for determining acetoside in rat plasma and to investigate the pharmacokinetic characteristics of acetoside in rats.

METHODSix rats were orally administered with 150 mg x kg(-1) acetoside and their blood samples were collected at different time points. The plasma concentration of acetoside was determined by reserved HPLC, and the pharmacokinetic parameters were calculated by DAS 2.0 software.

RESULTThe regression equation of acetoside in rats plasma was Y = 3.509 8X-0.096 8 (r = 0.996 8), which showed a good linear relation at 0.125-2.5 mg x L(-1). The method showed a recovery of more than 85%, and both inter-day and intra-day RSDs were less than 15%. After the oral administration of 150 mg x kg(-1) acetoside, the concentration-time curves of acetoside were expressed in a open two-compartment model. The main pharmacokinetics parameters of T(max), C(max), t(1/2alpha), t(1/2beta), AUC(0-t), AUC(0-infinity), CL/F, V/F and K(a) were respectively 0.36 h, 1.126 mg x L(-1), 0.759, 4.842 h, 3.134, 3.766 mg x h x L(-), 87.089 L x h(t) x kg(-1), 207.704 L x kg(-1) and 6.345 h(-1) respectively.

CONCLUSIONIt is first time to establish such a HPLC method to determine the concentration of acetoside in plasma. The method is so highly specified and sensitive that it can ble used in quantitative analysis in vivo on acetoside.

Animals ; Chromatography, High Pressure Liquid ; Female ; Glucosides ; chemistry ; pharmacokinetics ; Male ; Phenols ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVEThe pathogenesis of minimal change nephrotic syndrome (MCNS) remains unclear. This study aimed to investigate the expression of interleukin-6 (IL-6) in rats with doxorubicin-induced nephropathy and its possible roles in the pathogenesis of MCNS.

METHODSEighty-three male Wistar rats were randomly assigned into a control group (n=32) and a nephropathy group (n=51). Nephropathy was induced by a single tail vein injection of doxorubicin (5 mg/kg). The control group was injected with normal saline. Twenty-four-hour urinary protein excretion was measured 7, 14, 28 and 42 days after doxorubicin injection. IL-6 expression in urine and renal tissues was determined using ELISA 7, 14, 28 and 42 days after doxorubicin injection.

RESULTSThe urinary protein excretion increased significantly in the nephropathy group 7, 14, 28 and 42 days after doxorubicin injection compared with that in the control group (P<0.01). IL-6 expression in urine and renal tissues increased significantly 7, 14, 28 and 42 days after doxorubicin injection compared with that in the control group (P<0.01). IL-6 expression in urine and renal tissues was positively correlated with 24-hour urinary protein excretion in the nephropathy group (r=0.794, P<0.01; r= 0.870, P<0.01). IL-6 expression in urine was positively correlated with that in renal tissues (r=0.739, P<0.01).

CONCLUSIONSIL-6 expression in the urine and renal tissues is increased in MCNS rats. IL-6 might play an important role in the pathogenesis of MCNS.

Animals ; Antibiotics, Antineoplastic ; toxicity ; Disease Models, Animal ; Doxorubicin ; toxicity ; Interleukin-6 ; analysis ; Kidney ; chemistry ; Male ; Nephrosis, Lipoid ; chemically induced ; immunology ; Rats ; Rats, Wistar

Objective:The susceptibility of tinnitus rats with low estrogen level induced by sodium salicylate and the changes of tumor necrosis factor α (TNF-α) in serum were observed to investigate the relationship between tinnitus occurrence and estrogen level.Methods:Forty-two healthy female Wistar rats were randomly divided into control group( n=6), normal group( n=6), sham operation group( n=6) and ovariectomized group( n=24). Control group was intraperitoneally injected with normal saline 200 mg/kg for 14 consecutive days. Normal group, sham operation group and ovariectomized group were intraperitoneally injected with sodium salicylate 200 mg/kg for 14 consecutive days. Before and after sodium salicylate induction, the tinnitus behavior of rats in each group was detected by prepulse inhibition (PPI) and gap pre-pulse inhibition of the acoustic startle (GPIAS) test. Before and after sodium salicylate induction, blood samples were collected from eyeballs of rats in each group, and serum levels of estradiol and TNF-α were detected by ELISA. SPSS 25.0 software was used to analyze the data. Results:(1) Following 14 days of sodium salicylate intervention, there was no significant difference in PPI inhibition rate between groups or within groups(all P>0.05). (2)There was no significant difference in the inhibition rate of GPIAS in the four groups before sodium salicylate injection( F=0.217, P>0.05). With sodium salicylate injected for 14 days, the inhibition rate of GPIAS in ovariectomized group (30.88%±15.40%) was significantly lower than that in the other three groups (44.11%±21.06%, 38.27%±10.92%, 51.59%±11.34%), and the difference was statistically significant( F=3.533, P<0.05). The inhibition rate of GPIAS in ovariectomized group with sodium salicylate injected for 14 days was significantly lower than that before injection, and the difference was statistically significant( t=2.977, P<0.05).There was no significant difference in GPIAS inhibition rate between the other three groups before and after sodium salicylate injection( P>0.05). (3)The level of TNF-α in ovariectomized rats was significantly higher than that in the other three groups, the difference was statistically significant(all P<0.05). With sodium salicylate injection for 14 days, TNF-α level in the ovariectomized group increased more significantly than that in the other three groups, the difference was statistically significant( F=8.045, P<0.05). TNF-α levels increased following salicylate injection in normal group, sham operation group and ovariectomized group, and the differences were statistically significant( t value was -4.843, -4.932 and -5.965 respectively, each P<0.05). There was no significant difference in TNF-α levels before and after normal saline injection in control group(all P>0.05). Conclusion:Low estrogen levels increase susceptibility to sodium salicylate-induced tinnitus. Decreased estrogen levels may increase susceptibility to tinnitus through the increased expression of pro-inflammatory factor TNF-α.

Objective To explore the feasibility and advantage of fluoroscope in identification of brain structures in nude mice with green fluorescent protein (GFP) expression. Methods We laid the whole brain separated from 8-week adult nude mice with GFP expression into SLY mouse brain blocker to produce slices of 1 or 0.9 mm thickness; and then,25 μm-thickness frozen sections were cut.Fluoroscope was employed to observe the morphological structure to define their anatomic structures with reference to The Mouse Brain in Stereotaxic Coordinates compiled by Paxinos. After the observation,these frozen sections were performed Nissi staining for contrast. Results Different structures can be identified by their distinct fluorescence intensity:the dense areas of nuclei,Nissl bodies and nerve tract showed low fluorescence intensity; while the structures around the areas of nuclei and nerve tract,such as,the plexiform layer of olfactory bulb and the molecular layer of cerebella,showed high fluorescence intensity.The fluorescence intensity was attenuated obviously after Nissl staining; the visualized structural information observed under stereomicroscope was in accordance with that viewed by fluoroscope.Conclusion The identification of brain structure in nude mice with GFP by fluoroscope can serve as an experimental platform being applied in the anatomic structure positioning in fluorescence tracer experiments.

Honokiol is an active compound purified from magnolia that has been shown to induce cell differentiation, apoptosis, and anti-angiogenesis effects, as well as an enhancement in tumor growth delay in combination with chemotherapeutic agents in several mouse xenograft models. Our goal was to investigate the radiosensitization effect of honokiol on lung carcinoma. The radiosensitization effect of liposomal honokiol in Lewis lung carcinoma cells (LL/2) was analyzed using an in vitro clonogenic survival assay. For an in vivo study, Lewis lung carcinoma-bearing C57BL/6 mice were treated with either liposomal honokiol at 25 mg/kg or 5 Gy of single tumor radiation, or a combination of both over 12 days of treatment. The tumor growth delay and the survival time were evaluated. In addition, histological analysis of tumor sections was performed to examine changes by detecting the microvessel density and apoptosis in tumor tissues. In the clonogenic survival assay, LL/2 cells treated with IC50 Lipo-HNK for 24 h showed a radiation enhancement ratio of 1.9. After 12 days of combination treatment, the tumor volume decreased 78% and produced an anti-tumor activity 1.3-fold greater than a predicted additive effect of honokiol and radiation alone. This combination treatment also caused an 8.7 day delay in tumor growth. The cell cycle distribution and histological analysis demonstrated that liposomal honokiol has an anti-tumor effect via inducing apoptosis and inhibiting angiogenesis. Liposomal honokiol can enhance tumor cell radiosensitivity in vitro and in vivo, indicating that radiotherapy combined with liposomal honokiol can lead to greater anti-tumor efficacy.
Angiogenesis Inhibitors/administration & dosage/*therapeutic use ; Animals ; Apoptosis ; Biphenyl Compounds/administration & dosage/*therapeutic use ; Carcinoma, Lewis Lung/drug therapy/radiotherapy/*therapy ; Cell Cycle/drug effects/radiation effects ; Cell Line, Tumor ; Combined Modality Therapy ; Humans ; Lignans/administration & dosage/*therapeutic use ; Liposomes ; Lung Neoplasms/drug therapy/radiotherapy/*therapy ; Magnolia/chemistry ; Mice ; Neoplasm Transplantation ; Neovascularization, Pathologic/drug therapy/radiotherapy ; Radiation Tolerance ; Transplantation, Heterologous