ABSTRACT
ObjectiveTo explore the role of transient receptor potential vanilloid 1 （TRPV1） channel in reducing cardiomyocyte toxicity of Aconiti Kusnezoffii Radix processed with Chebulae Fructus. MethodH9c2 cardiomyocytes cultured in vitro were used as a model to assess cell viability by methyl thiazolyl tetrazolium （MTT） assay， the expression of TRPV1 mRNA was detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and the leakage rate of lactate dehydrogenase （LDH）， the changes of nucleus， reactive oxygen species （ROS）， mitochondrial membrane potential and Ca2+ contents were detected by enzyme linked immunosorbent assay （ELISA）. ResultCompared with the blank group， when the concentration was ≥0.5 g·L-1， the cell viability was significantly decreased （P<0.01）， the leakage rate of LDH， the release of ROS and Ca2+ were increased， the mitochondrial membrane potential was decreased， and the nucleus was pyknosis or even broken in raw Aconiti Kusnezoffii Radix and Aconiti Kusnezoffii Radix processed with Chebulae Fructus groups. When the concentration was ≥0.5 g·L-1， compared with the same mass concentration of raw Aconiti Kusnezoffii Radix group， the cell viability increased significantly （P<0.01）， the leakage rate of LDH， the release of ROS and Ca2+ decreased， the mitochondrial membrane potential increased， and the nuclear morphology improved in Aconiti Kusnezoffii Radix processed with Chebulae Fructus group. Application of the same mass concentration of raw Aconiti Kusnezoffii Radix to H9c2 cardiomyocytes pretreated with the TRPV1 inhibitor BCTC significantly increased cell viability， decreased leakage rate of LDH， ROS and Ca2+ release， increased mitochondrial membrane potential and improved nuclear pyknosis compared with untreated H9c2 cardiomyocytes. Application of the same mass concentration of Aconiti Kusnezoffii Radix processed with Chebulae Fructus to H9c2 cardiomyocytes pretreated with BCTC decreased cell viability， increased LDH leakage rate， ROS and Ca2+ release， reduced mitochondrial membrane potential compared with untreated H9c2 cardiomyocytes. Real-time PCR results showed that both raw Aconiti Kusnezoffii Radix and Chebulae Fructus decoction could increase the expression of TRPV1 mRNA in cardiomyocytes in a concentration dependent manner. ConclusionRaw Aconiti Kusnezoffii Radix can induce cardiomyocyte apoptosis and cardiotoxicity by activating TRPV1 channel， while Aconiti Kusnezoffii Radix processed with Chebulae Fructus can attenuate the toxicity through TRPV1 channel， which may be related to the synergistic effect of acid components in Chebulae Fructus and alkaloids in Aconiti Kusnezoffii Radix on TRPV1 channel.
ABSTRACT
OBJECTIVE@#To investigate the effects of puerarin on the activity of superoxide dismutase (SOD), and expressions of advanced glycation end-product (AGE) receptor (RAGE) and vascular endothelial growth factor (VEGF) in retinas of streptozotocin (STZ)-induced early diabetic rats.@*METHODS@#Diabetic rat models were established by inducing diabetes via intra-peritoneal injection of STZ. Rats were randomly divided into normal (control), diabetic (DM), and DM+ puerarin groups. After intra-gastric administration of puerarin (500 mg/kg/day for 4 weeks), levels of SOD and malondialdehyde (MDA) were determined in serum and retina. mRNA and protein expression levels of RAGE and VEGF in retinas were determined by real-time polymerase chain reaction (RT-PCR) (mRNA) and Western blot analysis (protein levels).@*RESULTS@#There was significantly lower SOD activity and significantly higher MDA in serum and retinas of the DM group compared with the two other groups (P<0.05). After treatment with puerarin, SOD activity increased and MDA content decreased in this group (P<0.05). mRNA and protein expression levels of RAGE and VEGF in the DM group were significantly higher than those of the other groups (P<0.05), and decreased after puerarin treatment (P<0.05).@*CONCLUSIONS@#Puerarin is able to enhance SOD activity, and inhibit RAGE and VEGF expressions in retinas of STZ-induced early diabetic rats.
Subject(s)
Animals , Male , Rats , Blotting, Western , Diabetes Mellitus, Experimental , Drug Therapy , Pathology , Enzyme Activators , Gene Expression Profiling , Isoflavones , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Receptor for Advanced Glycation End Products , Receptors, Immunologic , Retina , Pathology , Superoxide Dismutase , Metabolism , Treatment Outcome , Vascular Endothelial Growth Factor AABSTRACT
<p><b>OBJECTIVE</b>To explore changes in structure and function of the mitochondria of human gastric carcinoma BGC-823 cells after Newcastle disease virus (NDV) infection.</p><p><b>METHODS</b>Electron microscopy was applied to observe the structure of mitochondria; Rhodamine 123 staining was used to determine the mitochondrial membrane potential; the activity of Na(+)-K(+)-ATPase and Ca(2+)-ATPase were also determined and the release of cytochrome C was detected by Western blotting.</p><p><b>RESULTS</b>The structure of mitochondria in the tumor cells infected with NDV changed distinctly. In the infected group the activity of mitochondrial Na(+)-K(+)-ATPase and Ca(2+)-ATPase significantly declined (P < 0.01), and compared with control cells, mitochondrial trans-membrane potential was decreased. NDV infection induced the decrease of cytochrome C levels.</p><p><b>CONCLUSION</b>The effects of NDV infection on the structure and functions of mitochondria of human gastric carcinoma BGC-823 cells might play a role in the oncolysis of NDV.</p>
Subject(s)
Animals , Chick Embryo , Humans , Carcinoma , Metabolism , Virology , Cell Line, Tumor , Cytochromes c , Metabolism , Membrane Potential, Mitochondrial , Mitochondria , Metabolism , Virology , Newcastle Disease , Metabolism , Virology , Newcastle disease virus , Physiology , Sodium-Potassium-Exchanging ATPase , Metabolism , Stomach Neoplasms , Metabolism , VirologyABSTRACT
Objective: To investigate the effects of selective inhibition of HER 2 expression by Newcastle disease virus to the proliferation of human gastric adenocarcinoma cell line BGC-823. Methods: The expression of HER 2 mRNA and protein in Newcastle disease virus-infected gastric adenocarcinoma cells was determined by RT-PCR and Western blotting. The inhibitory effect of Newcastle disease virus on proliferation of BGC-823 cells were assessed by MTT assay. Apoptotic rate of BGC-823 cells was measured by flow cytometry (FCM). Results: The infection of BGC-823 cells with Newcastle disease virus successfully knocked down the expression of HER 2 at mRNA and protein levels. Down-regulation of HER 2 gene inhibited the proliferation and increased the apoptotic rate of BGC-823 cells compared with the control group (P < 0.01). Conclusions: Infection with Newcastle disease virus efficiently killed BGC-823 cells and induced apoptosis. The mechanism may be due to the inhibition of HER 2 gene expression in BGC-823 cells.