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1.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 436-445, 2020.
Article in English | WPRIM | ID: wpr-827226

ABSTRACT

This study investigated the effects of X-ray irradiation on primary rat cardiac fibroblasts (CFs) and its potential mechanism, as well as whether sodium tanshinone IIA sulfonate (STS) has protective effect on CFs and its possible mechanism. Our data demonstrated that X-rays inhibited cell growth and increased oxidative stress in CFs, and STS mitigated X-ray-induced injury. Enzyme-linked immuno-sorbent assay showed that X-rays increased the levels of secreted angiotensin II (Ang II) and brain natriuretic peptide (BNP). STS inhibited the X-ray-induced increases in Ang II and BNP release. Apoptosis and cell cycle of CFs were analyzed using flow cytometry. X-rays induced apoptosis in CFs, whereas STS inhibited apoptosis in CFs after X-ray irradiation. X-rays induced S-phase cell cycle arrest in CFs, which could be reversed by STS. X-rays increased the expression of phosphorylated-P38/P38, cleaved caspase-3 and caspase-3 as well as decreased the expression of phosphorylated extracellular signal-regulated kinase 1/2 (ERK 1/2)/ERK 1/2 and B cell lymphoma 2 (Bcl-2)/Bcl-2 associated X protein (BAX) in CFs, as shown by Western blotting. STS mitigated the X-ray radiation-induced expression changes of these proteins. In conclusion, our results demonstrated that STS may potentially be developed as a medical countermeasure to mitigate radiation-induced cardiac damage.

2.
Chinese Journal of Applied Physiology ; (6): 259-262, 2012.
Article in Chinese | WPRIM | ID: wpr-329893

ABSTRACT

<p><b>OBJECTIVE</b>To get stable cell line expressing B domain-deleted human FVIII (BDDhFVIII) by constructing the eukaryotic expression plasmid.</p><p><b>METHODS</b>Eukaryotic expression plasmid containing BDDhFVIII was constructed and transfected into HepG2 cells via electroporation. The expression and purification of the target protein was detected by Western blot.</p><p><b>RESULTS</b>Results of enzyme digestion and sequence analysis demonstrated that the gene of BDDhFVIII was correctly inserted into the eukaryotic expression vector pcDNA4/v5-his. Western blot confirmed the successful expression of BDDhFVIII at the protein levels in HepG2 cells.</p><p><b>CONCLUSION</b>The constructed eukaryotic expression vector was able to generate high level expression of human FVIII in HepG2 cells, thus could construct human blood coagulation FVIII stable cell line successfully.</p>


Subject(s)
Humans , Electroporation , Factor VIII , Genetics , Gene Expression , Genetic Vectors , Hemophilia A , Genetics , Hep G2 Cells , Plasmids , Recombination, Genetic
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