ABSTRACT
This study aimed to investigate the effect of methyl eugenol(ME) on hypoxia/reoxygenation(H/R)-induced injury of human renal tubular epithelial HK-2 cells and its mechanism. The viability of HK-2 cells cultured with different concentrations of ME and exposed to H/R was detected by cell counting kit-8(CCK-8) assay. The effect of ME on the morphology of HK-2 cells was observed under an inverted microscope. The content of intracellular reactive oxygen species in different groups was detected after 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA) fluorescence staining. Cell apoptosis was determined by flow cytometry. Changes in mitochondrial membrane potential were monitored by JC-1 dye. The concentrations of nuclear factor erythroid 2 related factor 2(Nrf2), heme oxygenase-1(HO-1), and nicotinamide adenine dinucleotide phosphatase oxidase 4(Nox4) were measured by Western blot, followed by the assay of Nrf2 concentration changes in cytoplasm and nucleus by confocal fluorescence staining. The results showed that when the concentration of ME was 0-40 μmol·L~(-1), the activity of HK-2 cells was not affected. Compared with the model group, ME enhanced the activity of HK-2 cells and the cell morphology was normal. As revealed by further experiments, ME inhibited the release of reactive oxygen species and the decline in mitochondrial membrane potential of HK-2 cells after H/R injury, promoted Nrf2/HO-1 expression and Nrf2 translocation to the nucleus, and down-regulated the expression of Nox4, thereby significantly reducing apoptosis. This protective effect of ME could be reversed by the specific Nrf2 inhibitor ML385. These findings have preliminarily proved that ME effectively protected HK-2 cells against H/R injury, which might be related to its promotion of Nrf2/HO-1 signaling pathway and inhibition of Nox4. Such exploration on the possible mechanism of ME in the treatment of renal ischemia-reperfusion injury(IRI) and protection of organ function from the perspective of antioxidant stress has provided reference for related research on the treatment of acute kidney injury with traditional Chinese medicine.
Subject(s)
Humans , Apoptosis , Epithelial Cells/metabolism , Eugenol/pharmacology , Heme Oxygenase-1/metabolism , Hypoxia , NF-E2-Related Factor 2/metabolism , Oxidative Stress , Reactive Oxygen Species , Reperfusion Injury/drug therapyABSTRACT
At present,the mechanism of breathing trainer is not very clear,but it mainly bases on the neuromuscular plasticity of re-spiratory muscle.The breathing trainers appeared on the market mainly included resistance load or threshold load breath-ing trainers,abdominal breathing trainers,multifunctional breathing trainers,and new breathing trainers.However,the efficacy of breathing trainer on the stable patients with chronic obstructive pulmonary disease remains uncertain, and needs further researches.
ABSTRACT
<p><b>OBJECTIVE</b>To study the feasibility of using tetracysteine (TC) reporter in gene therapy.</p><p><b>METHODS</b>Effects of TC reporter and conventional reporter genes encoding green fluorescence protein (GFP) and luciferase (Luc) on expression and function of the therapeutic gene MGMT(P140K) were compared. Cytotoxicity and drug resistance were studied by Western blot. TC reporter used in therapy was analyzed by flow cytometry (FCM).</p><p><b>RESULTS</b>The TC reporter had no toxicity to cells and neither affected the expression or activity of therapeutic gene as compared to GFP and Luc. TC could be used in blood sample detection.</p><p><b>CONCLUSION</b>TC is a new kind of reporter gene for lentiviral vector in future gene therapy.</p>
Subject(s)
Animals , Cricetinae , Humans , CHO Cells , Cricetulus , Cysteine , Genetics , Metabolism , Gene Expression Regulation , Genes, Reporter , Genetic Therapy , Lentivirus , Genetics , Lymphocytes , MetabolismABSTRACT
To introduce the development of myelodysplastic syndrome network course.
ABSTRACT
Objective To compare the effects of curcumin and enalapril on transdifferentiation of renal tubular epithelial cells in unilateral ureteral obstruction (UUO) rats. Method UUO rat model was used. Male wistar rats were randomly divided into four groups with 8 rats each:shame-operated group (0.9% saline solution), model group (dimethyl sulphoxide), curcumin treatment group [curcumin 100 mg/(kg?d)], and enalapril treatment group [enalapril 10 mg/(kg?d)]. Medicines were given intraperitoneally daily two days after the operation. All rats were killed 4w after surgery. The degree of tubulointerstitial fibrosis was scored by HE and Masson staining, and the sites and levels of protein expression of TGF-?1、?-SMA and vimentin were detected by immunohistochemistry staining. Result Compared with shame-operated group, the serum urea nitrogen level was significantly increased (P 0.05). Conclusion The pharmacological effects of curcumin and enalapril may be partly mediated by inhibiting the transdifferentiation of renal tubular epithelial cells towards myofibroblast in UUO rats.
ABSTRACT
Objective To investigate the effect of curcumin on renal interstitial fibrosis following unilateral ureteral obstruction (UUO), and compare the effects of Curcumin with that of Enalapril. Method UUO rat model was used. Male Wistar rats were randomly divided into four groups with 8 rats each:shame-operated group (0.9% saline solution), model group (dimethyl sulphoxide, DMSO), Curcumin treatment group [curcumin 100 mg/(kg?d)], and Enalapril treatment group [enalapril 10 mg/(kg?d)]. All rats were killed 4 w after surgery. The degree of tubulointerstitial fibrosis was scored by HE and Masson staining. The sites and levels of protein expression of TGF-?1, CTGF and TIMP-1 were detected by immunohistochemistry staining. Result Compared with those of shame-operated group, the blood urea nitrogen level was significantly increased (P 0.05). Conclusion Pharmacological effects of Curcumin or Enalapril on unilateral ureteral obstruction rats are quite similar. These effects may result from the down-regulation of TGF-?1, CTGF and TIMP-1 expression in kidney tissue.