ABSTRACT
AIM To explore the effects of glycyrrhetinic acid on the gastric ulcer rats infected by Helicobacter pylori (Hp) and its action mechanism.METHODS Gastric ulcer rat models were induced by acetic acid stress and then followed by Hp infection.After treatment with low and high doses of glycyrrhetinic acid,the ulcer index,gastric acid and proteinase activities in gastric ulcer rats were analyzed.The effects of glycyrrhetinic acid on the expressions of BCL2 and Caspase-3,the GSK3β activity in gastric mucosa and gastric epithelial cells,and the cell apoptosis level were then detected.RESULTS Glycyrrhetinic acid reduced the ulcer index,gastric acid and proteinase activities in rats.Besides,the expression of BCL2 was significantly up-regulated by glycyrrhetinic acid in gastric mucosa and gastric epithelial cells,whereas the expression of Caspase-3,level of cell apoptosis,and GSK3β activity were significantly reduced.After the treatment with GSK3 β activator LY294002,the level of BCL2 was down-regulated,Caspase-3 expression was increased,and the level of cell apoptosis was enhanced.CONCLUSION Glycyrrhetinic acid promotes the healing of gastric ulcer infected by Hp via regulating GSK3β activity and inhibiting apoptosis of gastric epithelial cells.
ABSTRACT
[ ABSTRACT] AIM:To investigate the effect of artemisinin on lipopolysaccharide ( LPS)-induced intestinal epi-thelial barrier damage in IEC-6 cells and its molecular mechanism.METHODS:Cultured IEC-6 cells were divided to 5 groups:control group, LPS (100 mg/L) group and LPS +Artemisinin (30, 50 and 100μmol/L) groups.The cytotoxici-ty was detected by MTT assay.The releases of TNF-α, IL-1βand IL-6 in the IEC-6 cells were measured by ELISA.The transepithelial electrical resistance ( TER) was detected by electrical resistance tester, and the horseradish peroxidase (HRP) flux permeability were analyzed by a microplate reader.The expression of tight junction proteins, ZO-1, claudin-1 and occludin, and the expression of TLR4/MyD88/NF-κB at mRNA and protein levels were determined by RT-qPCR and Western blot.RESULTS:Artemisinin alone (up to 100 μmol/L) or in combination with LPS (100 mg/L) was not toxic to IEC-6 cells.Compared with control group, the releases of TNF-α, IL-1βand IL-6 in the culture supernatant of IEC-6 cells significantly increased after treatment with LPS.The expression of TLR4/MyD88/NF-κB was activated by LPS.LPS down-regulated the protein expression of ZO-1, claudin-1 and occludin.However, artemisinin treatment decreased the re-leases of TNF-α, IL-1βand IL-6 in the culture supernatant of IEC-6 cells.The expression of TLR4/MyD88/NF-κB at mR-NA and protein levels was gradually reduced after treatment with artemisinin.In addition, artemisinin upregulated the pro-tein expression of ZO-1, claudin-1 and occludin significantly (P<0.01) in a dose-dependent manner.CONCLUSION:Artemisinin attenuates LPS-induced intestinal epithelial barrier damage by inhibiting TLR4/MyD88/NF-κB activation in the IEC-6 cells.
ABSTRACT
Objective To explore the function of MDA on diabetic nephropathy .Methods Glomerular mesangial cells ( GMC) were pretreated with MDA at a final concentrations of 0μmol/L, 1μmol/L, 5μmol/L, 10μmol/L and 50 μmol/L.MTT assay was used to examine the viability of GMC ) .AnnexinV-FITC was used to evaluate effect of MDA on cell apoptosis .RT-PCR and western blot were used to analyze the expression of Nrf 2, HO-1 andγGCL.Results MDA treatment inhibited GMC viability in a dose-dependent manner .MDA at the concentration of more than 5 μmol/L induced mass production of ROS in GMC ( P<0.05 ) .In addition , antioxygen of tBHQ may relieve MDA-induced reduction of cell viability .MDA inhibited the expression of HO-1 , γGCLand Nrf2 ( P <0.05 ) .Conclusions MDA inhibites GMC viability and promotes the cell apoptosis by ROS production through in-hibiting Nrf2/HO-1-γGCL.
ABSTRACT
Objective To obtain recombinant human stanniocalcin 1 ( STC1 ) with biological activity in Escheri.coli cells expression.Methods The gene was cloned into pET32b( +) vector by fused with thioredoxin and His tag .E.coli BL21(DE3) competent cells were transfomed by the recombinant vector .After renaturation, the fusion protein was digested with thrombin and intact STC1 protein was purified from the digested protein using Ni ion affinity chromatography .Recombi-nant humanSTC1 protein was confirmed by Western blot analysis using goat anti-STC1 antibody.The biological activity of STC1 in rat was assayed using standard method for assessment of renal function .Results The recombinant human STC 1 fu-sion protein is successfully expressed in Escherichia coli, the fusion protein was purified by affinity chromatography from the inclusion body and renaturated .Intact hSTC1 protein was released by thrombin digestion and purified by Ni ion affinity col-umn.The intact STC1 proteins was confirmed by Western blot analysis .Rat bioassay revealed that STC1 boosted phosphate reabsorption.Conclusion Recombinant STC1 protein was successfully expressed and has native biological activities .This protein could be used as an antigen for the preparation of monoclonal antibody against humanSTC 1.
ABSTRACT
Objective: To study the distribution,combination law of syndrome factors and characteristics of base syndromes of pneumonia based on document data.Methods: Combined methods of computer retrieval and man-made retrieval were adopted,the document data from 1981 to 2004 on lower respiratory tract infection was gathered by way of document database index.And the data was analyzed by description,Logistic regression and clustering analysis.Results: 32 documents and 141 items were used for analysis.The results indicated that among the 17 syndrome factors related to syndrome differentiation of pneumonia,the highest frequency of location syndrome factor was lung;the frequency of f ire,heat and yin-def iciency was higher;pathogenic wind,pathogenic heat and phlegm were the main etiological factors.In the document study,there were four patterns in the combinative model of syndrome factors,the combination of two syndrome factors and the combination of three syndrome factors were the main patterns,the cumulative percentage was 95.7%.Meanwhile,the main symptoms and the sub-symptoms of the mainly syndromes were screened out by adopting statistical description,logistic regression and clustering analysis.Conclusion: The main syndrome types and their inscapes of pneumonia can provide enough references for the next step research.