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<p><b>OBJECTIVE</b>To study the inhibitory effect of 10-gingerol on the proliferation of hepatocellular carcinoma HepG2 cells and the role of Src/STAT3 signaling pathway in mediating the effect.</p><p><b>METHODS</b>SYBYL-X2.1 software was used to simulate the interaction between 10-gingerol and Src. HepG2 cells treated with 10-gingerol at 1, 3, 10 or μol/L for 24 h were assessed for cell viability using MTT assay, and EdU staining was used to detect the cell proliferation and calculate the number of positive cells. The expressions of p-Src and p-STAT3 were detected using Western blotting, and the mRNA expressions of the target genes of STAT3 (cyclin D1 and CMCC) were detected using qPCR.</p><p><b>RESULTS</b>10-gingerol was capable of forming hydrogen bond with such Src residues as TRY-340, MET-341, MET-314, ASP-404, and ILE-336. MTT assay showed that 10-gingerol at 3 and 10 μmol/L significantly lowered the viability of HepG2 cells ( < 0.001). Treatment with 1, 3, and 10 μmol/L 10-gingerol significantly reduces the number of EdU-positive HepG 2 cells ( < 0.001). Western blotting showed that 10-gingerol at 3 and 10 μmol/L significantly decreased the phosphorylation levels of Src and STAT3 in HepG2 cells ( < 0.01). 10-gingerol at 1, 3, and 10 μmol/L significantly decreased the mRNA expressions of cyclin D1 and CMCC as shown by qPCR ( < 0.01).</p><p><b>CONCLUSIONS</b>10-gingerol can dose-dependently inhibit the proliferation of HepG2 cells and suppress the activation of Src and STAT3.</p>
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Objective To observe the influence of Buyang Huanwu Decoction ( BYHWD) on the inhibition of rat myocardial H9C2 cell activity and SH2-containing tyrosine phosphatase-1 ( SHP-1) activity induced by trastuzumab, and to explore the possible regulatory mechanism after observing the intervention of BYHWD on rat myocardial H9C2 cell transfected with SHP-1 or SHPC/S-1 gene. Methods The eukaryotic expression vectors pcDNA3.1 (+)- SHP-1 and pcDNA3.1 (+) -SHPC/S–1 were constructed and then were transfected to rat myocardial H9C2 cells using the method of liposome transfection. The cells with positive clones were screened out with G418, and then were cultured with trastuzumab for maintaining growth. Using quantitative RT-PCR, we detected the expression of SHP-1 gene and SHPC/S - 1 gene in rat myocardial H9C2 cells. The phosphatase activity analysis was used for observing the regulatory effect of BYHWD on SHP-1 in myocardial cells. Furthermore, we observed the apoptosis of rat myocardial H9C2 cells by methyl thiazolyl tetrazolium (MTT) assay after treatment with BYHWD. Results Sequencing results indicated the successful construction of eukaryotic expression vectors, which had stable expression in myocardial H9C2 cells even under the intervention of trastuzumab. The results of phosphatase analysis showed that H9C2-SHP-1 had the highest activities of phosphatase, but the activities were decreased after the intervention with BYHWD ( P<0.05) . The results of MTT assay also showed the apoptotic rate of H9C2-SHP-1 cells was decreased after treatment with BYHWD ( P <0.05) . Conclusion BYHWD can promote the proliferation of myocardial H9C2 cells inhibited by trastuzumab, and can regulate the expression of SHP-1 in myocardial cells, which will supply reference to the further study of treatment of trastuzumab-induced cardiac toxicity.
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Objective To investigate the inhibitory mechanism of emodin on lipopolysaccharide(LPS)-induced nitric oxide(NO)generation in rat peritoneal macrophages.Methods NO production and iNOS expression were measured through nitrite assay and Western blotting assay,respectively.NF-kB activity and nuclei P65 expression were estimated by dual-luciferase and Western blotting assay,respectively.Intracellular free Ca2+([Ca2+]i)was detected using the ratiometric fluorescent calcium indicator dye,Fura-2,and a microspectrofluorometer.PLC-γphosporylation was analyzed by Western blotting assay.Results First,emodin was found playing active roles in suppressing LPS-induced NF-kB activation in rat peritoneal macrophages.Second,emodin down-regulated transient[Ca2*]i and could increase in NF-kB upstream signal.Finally,emodin suppressed phosphorylation of PLC-γ by LPS stimulation in the upstream of[Ca2+]i.Conclusion Suppression of PLC-γ phosphorylation is involved in emodin inhibiting NO generation by LPS stimulation in rat peritoneal macrophages.
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Objective To explore the relationship between polymorphism of G-protein ?3 subunit (GNB3) gene and the emotion trait of chest stuffiness and pains patient. Methods The GNB3 gene type in 27 patients of depression of chest stuffiness and pains, 40 patients of no depression of chest stuffiness and pains, and 20 patients of healthy subjects (control) was determined by polymerase chain reaction and restriction fragment incision enzyme, and score of depression was evaluated. Result Occurrence of depression in chest stuffiness and pains was significantly higher than that in control (P
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OBJECTIVE: Ventricular reconstruction after myocardial infarction is one of the important dangerous factors to the remote occurrence of cardiac functional disturbance. Chinese medicine is of unique advantages in the main therapeutic principles and methods for the post-infarct ventricular reconstruction.DATA SOURCES: To search for and proofread the relevant literatures on the researches on post-infarct ventricular reconstruction in Nuclear Journals of Chinese Medicine from January 1987 to December 2003 by www. google. com,Medline. Referring word: ventricular reconstruction,myocardial infarction,removing blood stasis and resolving phlegm,clearing heat and nourishing yin.STUDY SELECTION: Fifty-six papers on post-infarct ventricular reconstruction were selected, excluded the original literatures on non-random researches and included the original literatures on non-blind researches.DATA EXTRACTION: Of 56 papers,14 papers were deleted for they were repeated in the contents in different degrees. The rest 42 papers were classified and 12 papers of which were selected as the references.DATA SYNTHESIS: Myocardial infarction is the syndrome in which the root cause is deficiency and the symptoms are excess. The main therapeutic principles are to benefit qi,eliminate stasis,resolve phlegm,activate blood circulation,regulate qi,strengthen spleen,clear away heat and nourish yin. Of which, benefiting qi is the primary of the treatment for the root cause and nourishing yin is the secondary of the treatment for the root cause. Eliminating stasis and resolving phlegm is the primary of the treatment for symptoms,regulating qi and clearing away heat is the secondary of the treatment for symptoms. To benefit qi and activate blood circulation can improve blood dynamics to probably benefit the protection of ventricular reconstruction by reducing ventricular pressure and vascular resistauce of systematic circulation.CONCLUSION: After myocardial infarction, it happens inadaptable myocardial hypertrophy and external stromal accumulation and fibrosis of cardiac muscles. Long-term application of the principles for eliminating stasis,resolving phlegm,clearing away heat and nourishing yin can resist ventricular reconstruction and improve post-infarct cardiac function.