[Abstract] Objective To investigate the role of dihydromyricetin (DHM) in the treatment of ischemic stroke in rats, and to explore the effect of DHM on the expression of inflammasome. Methods The middle cerebral artery occlusion (MCAO) model was induced by endovascular suture method. The therapeutic effect and mechanism of DHM were investigated by Longa score, TTC staining, Nissl staining, immunohistochemical staining and Western bloting. Results After DHM treatment, the motor capacity of MCAO rats was significantly improved, the infarct volume was significantly reduced, the brain structure and neuron morphology were improved, and the expressions of nod-like receptor protein-3 (NLRP3) and interleukin-1(IL-1) decreased significantly. Conclusion DHM can down-regulate the expression of NLRP3 and thus reduces the cerebral infarction volume and improves neurobehavioral performance in MCAO rats.

OBJECTIVETo analyze the effects of Ling Qi Juan Gan capsule drug-containing serum at different concentrations on the platelet-derived growth factor (PDGF)-induced proliferative capabilities of and JAK2 and p-STAT3 protein expression in hepatic stellate cells (HSC) using an in vitro system.

METHODSTwenty-five Sprague-Dawley rats were randomly divided into five equal groups for intragastric administration of physiological saline (10 ml/kg; group A), Fufang Biejia Ruangan tablet solution (1.5 g/kg; group B), or Ling Qi Juan Gan capsule solution at low dose (2.125 g/kg; group C1), mid dose (4.25 g/kg; group C2), or high dose (8.5 g/kg; group C3). The post-administration serum isolated from each rat (200 ml/L) was used to treat the HSC-T6 cell line following induction by PDGF (10 ng/ml). At 24, 48 and 72 h post-exposure, the cells' proliferation was measured using the Cell Counting Kit-8 (CCK-8) colorimetric assay. In addition, at 24 h post-exposure the expression of JAK2 and p-STAT3 was measured by western blotting (expressed as grey scale intensity). Multiple group comparison of repeated measures data was made by one-way ANOVA with Student-Newman-Keuls post hoc test.

RESULTSCompared to group A, groups C2 and C3 had significantly higher inhibited proliferation at all post-exposure time points examined (24 h: A = 1.550 +/- 0.065, C2 = 1.335 +/- 0.106, C3 = 1.241 +/- 0.205; 48 h: A = 1.311 +/- 0.650, C2 = 1.090 +/- 0.106, C3 = 0.909 +/- 0.191; 72 h: A = 1.039 +/- 0.103, C2 = 0.719 +/- 0.116, C3 = 0.641 +/- 0.110, F = 36.292, all P less than 0.05); in contrast, compared to group A, group C1 showed no inhibition of proliferation at 24 h (1.522 +/- 0.128, P = 0.717) but showed significantly higher inhibition of proliferation at 48 h and 72 h (1.153 +/- 0.183 and 0.753 +/- 0.210, respectively, F = 36.292, P less than 0.05). Compared with group A, all Ling Qi Juan Gan capsule-containing serum-treated groups showed significantly lower expression of both JAK2 (A = 1.605 +/- 0.024 vs. C1 = 1.170 +/- 0.042, C2 = 0.842 +/- 0.036, C3 = 0.555 +/- 0.036, F = 43.091) and p-STAT3 (A = 1.401 +/- 0.030 vs. C1 = 1.229 +/- 0.025, C2 = 0.668 +/- 0.034, C3 = 0.630 +/- 0.026, F = 78.426, all P less than 0.01).

CONCLUSIONLing Qi Juan Gan capsule drug-containing serum can inhibit the proliferation of HSC-T6 cells in a dose-dependent manner and cause an overall decrease in the expression of JAK2 and p-STAT3 in activated HSC, thereby leading to a suppression of the JAK/STAT signal transduction pathway.

Animals ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Hepatic Stellate Cells ; drug effects ; metabolism ; Janus Kinase 2 ; metabolism ; Male ; Platelet-Derived Growth Factor ; Rats ; Rats, Sprague-Dawley ; STAT3 Transcription Factor ; metabolism ; Serum ; Signal Transduction ; drug effects

OBJECTIVETo study the role of serum from asphyxiated neonates in the inducement of human renal proximal tubular epithelial cells (HK-2) adhesion to neutrophils and possible mechanisms.

METHODSHK-2 cells were cultured randomly with 20% serum from neonates (1, 3, and 7 days after asphyxia), pyrrolidine dithiocarbamate (PDTC) or placebo. The activity of myeloperoxidase (MPO), an indicator of adhesion ability of HK-2 cells to neutrophils in suspensions, was detected by the biochemistry assay. Intercellular adhesion molecule-1 (ICAM-1) and nuclear factor-kappaB (NF-kappaB) of HK-2 cells were examined with the immunohistochemical staining.

RESULTSThe expression of MPO in the post-asphyxial 1-day serum treatment group were significantly higher than that in the PDTC treatment and the control groups as well as the post-asphyxial 3 and 7-day serum treatment groups (P<0.01). The expression of ICAM-1 and NF-kappaB in the post-asphyxial 1-day serum treatment group was also significantly higher than that in the other groups (P<0.01).

CONCLUSIONSSerum from asphyxiated neonates can induce HK-2 cell adhesion to neutrophils, possibly through activating NF-kappaB and increasing the synthesis and expression of ICAM-1 on the surface of renal tubular epithelial cells.

Asphyxia Neonatorum ; blood ; complications ; Cell Adhesion ; Cells, Cultured ; Humans ; Infant, Newborn ; Intercellular Adhesion Molecule-1 ; analysis ; biosynthesis ; Kidney Tubules ; pathology ; NF-kappa B ; analysis ; metabolism ; Neutrophils ; physiology

Carcinoma, Hepatocellular ; virology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Liver Neoplasms ; virology ; Mutation ; Trans-Activators ; genetics

OBJECTIVESalvia miltorrhiza Bunge (SMB) is a traditional Chinese herb, which is considered to promote blood flow and remove blood stasis. This study examined whether SMB can alleviate injury induced by hypoxia/reoxygenation (H/R) in human kidney proximal tubular cells-2 (HK-2 cells).

METHODSThere were 3 experimental groups: control, H/R injury and SMB-treated H/R injury. H/R injury of HK-2 cells was induced by first covering the cells with and then removing liquid paraffin wax. Different concentrations of compound SMB solution (0.05%, 0.10%, 0.15% or 0.20%) were administered to the SMB-treated H/R injury group before the hypoxic injury. After 4, 12 and 24 hrs of hypoxia and 4, 12, 24 and 48 hrs of reoxygenation, morphologic changes of HK-2 cells were observed under an inverted microscope. Cell viability was measured by the MTT method. Lactate dehydrogenase (LDH) activity in the culture supernatants was assayed using biochemical methods; TNF-alpha levels were determined by radioimmunoassay (RIA).

RESULTSThe number of HK-2 cells was significantly reduced in the H/R injury group after hypoxia, and reached a nadir 24 hrs after hypoxia treatment. Various concentrations of SMB-treated groups showed significantly greater number of HK-2 cells than the H/R injury group. SMB solution (0.10%) produced the best effect. The levels of LDH and TNF-alpha in the H/R injury group were significantly increased, and reached a peak between 24 hrs of hypoxia and 4 hrs of reoxygenation when compared to the control group. Pre-treating with 0.10% SMB resulted in significantly lower levels of LDH and TNF-alpha than in the untreated H/R injury group at various time points of H/R.

CONCLUSIONSSMB has protective effects against H/R injury of HK-2 cells, possibly through inhibition of inflammatory cytokines.

Calcium ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Cytoprotection ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Kidney Tubules, Proximal ; drug effects ; pathology ; L-Lactate Dehydrogenase ; metabolism ; Plant Extracts ; pharmacology ; Salvia miltiorrhiza ; Tumor Necrosis Factor-alpha ; analysis

Objective To explore the role of Na~+/H~+ exchanger 1(NHE1) in injury of human renal tubular cells(HK-2) induced by postasphyxial-serum of neonates.Methods HK-2 was used as the target cell.The attacking concentration of postasphyxial-serum of neonates was 200 mL/L.First,the experiment was designed as control group and asphyxia group,the expression of NHE1 in the HK-2 was detected by immunohisto chemical method in the cells.Second,the experiment group was designed as control group,asphyxia group,and pretreatment with 5-N-Ethyl-N-isopropylamiloride(EIPA) group,then the change of morphology was observed under inverted microscope,and the cell viability was measured by methyl thiazolyl tetrazolium(MTT) method,and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with blank control group,the expression of NHE1 in the HK-2 was increased signi-ficantly in asphyxia group,the changes of morphology of HK-2 was most serious and obvious,the cell viability decreased,and the leakage rate of LDH increased significantly in asphyxia group.But compared with asphyxia group,the change of morphology of HK-2 was greatly improved,the cell injury was decreased obviously,the leakage rate of LDH was increased and viability was decreased in pretreatment group in a dose 2 ?mol/L.Conclusions The postasphyxial-serum may induce the expression of NHE1,which plays an important role in injury of renal tubular cell induced by postasphyxial-serum in neonates,and inhibiting activity of NHE1 may relieve the injury of renal tubular cells induced by postasphyxial-serum in neonates.

Objective To explore the protective effects of astragalus on injury of renal tubular cells(HK-2) induced by postasphyxial-serum from neonate.Methods Human renal proximal tubular cell line HK-2 cells were taken as subjects.The experiment was designed as:control group,model group,astragalus pretreatment group.The astragalus pretreatment group had 5 subgroups,which were pretreated for 24 hours.The serum of neonates who suffured asphyxia for 1 day,which concentration were 200 mL/L(volume fraction),were applied as attacking factor.The injury of morphology were observed under inverted microscope.The cell viability was measured by methyl thiazolyl tetrazolium methods.The leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,cell morphous changed,leakage rate of LDH elvated and cell survival rate decreased in model group,the variances were obviously significant(Pa

Objective To explore the effect of erythropoietin(EPO) on apoptosis of human renal tubular(HK-2) cells induced by postasphyxial-serum of neonates.Methods HK-2 cells were used as target cells.The experiment was divided into 4 groups,control group(n=8):HK-2 cells were maintained in standard medium;asphyxia group(n=8):HK-2 cells were treated with serum obtained from neonates with asphyxia.Each culture medium replaced with 200 mL/L suffocation DMEM/F12 serum culture medium;EPO pretreatment group(n=8):HK-2 cells were pretreated 24 h with serum contains 5?104 IU/L EPO,and then deal as asphyxia group;EPO and 5-hydroxydecanoic acid sodium salt(5-HD) pretreatment group(n=8): HK-2 cells were pretreated 24 h with serum contains 5?104 IU/L EPO and 500 ?mol/L 5-HD,and then deal as asphyxia group.All cells were cultured at 37 ℃ in humidified atmosphere with 50 mL/L CO2 for 24 h.The apoptosis rate of HK-2 cells was detected by flow cytometer.The expressions of Caspase-3 and X-linked inlnibitor of apoptosis protein(XIAP) of HK-2 cells were detected by using immunohistochemical method.Results Compared with control group,after stimulated with postasphyxial-serum,the apoptosis rate and the expression of Caspase-3 of HK-2 cells were significantly increased(Pa

Objective To investigate the role of postasphyxial-serum of neonate in inducing injury of human renal proximal tubular cells(HK-2 cells).Methods HK-2 cells were used as target cell.The neonatal different concentration postasphyxial-serum of 1,3,7 days after asphyxia were used as attacking means.The experimental groups were divided into 15 groups:the 2.5%,5.0%,10.0%,(20.0%) attacking concertration groups of 1,3,7 day after asphyxia and control group of each concertration.The culture medium and concertration of the control group and the experimental groups were the same.The changes of morphology were observed under inverted microscope,the cell viability was measured by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the changes in morphology of HK-2 were most serious and obvious,the cell viability were obviously decreased(all P

Adrenomedullin ; Animals ; Endothelin-1 ; blood ; Liver Cirrhosis, Experimental ; physiopathology ; Nitric Oxide ; blood ; Peptides ; blood ; pharmacology ; Portal Pressure ; drug effects ; Rats ; Rats, Wistar