Suspension Culture and Alkaloid Content of Pinelliae Rhizoma Tuber Under Stimulation of Salicylic Acid / 中国实验方剂学杂志

Objective: To study the suspension culture of tuber and its alkaloid content based on the stimulation of salicylic acid. Method: The tubers of Pinelliae Rhizoma in suspension tube were treated with different concentrations of exogenous salicylic acid to analyze the growth status. The content of alkaloids in tuber was detected by HPLC. Test conditions:chromatographic column for Agilent Eclipse plus C18 column (4.6 mm×250 mm,5 μm),mobile phase of acetonitrile water(4:96),the column temperature was maintained at 35℃,detection wavelength for inosine 250 nm,guanosine 260 nm,volume flow rate 1.0 mL·min-1. Result: The results showed that the exogenous salicylic acid had a certain effect on the growth of suspension tuber of Pinelliae Rhizoma. When the salicylic acid concentration was 150 μmol·L-1,the culture lasted for 25 days and the fresh weight reached the maximum value of 7.483 8 g. It also accumulates a certain amount of alkaloids. The linear range of guanosine was 0.03-0.45 μg (R2=0.999 6). After 10-days cultivatation in the salicylic acid concentration of 50 μmol·L-1,guanosine content of Pinelliae Rhizoma tubers reached a maximum of 1.353 3 mg·g-1. The linear range of inosine 0.003-0.045 μg (R2=0.999 5). When the salicylic acid concentration was 200 μmol·L-1,cultured for 30 days,the content of inosine in Pinelliae Rhizoma tubers reached the maximum value of 0.149 8 mg·g-1. Conclusion: The results of this experiment provide a reference for the study of tissue culture and rapid propagation of Pinelliae Rhizoma tubers and regulation of alkaloids,which are of great significance for the development of Pinelliae Rhizoma industry.

Effect of Menthol on Proliferation, Migration and Expressions of IL-8, CXCL-12 and VEGF in Hepatoma HepG2 Cells / 中国实验方剂学杂志

Objective: To study the effect of menthol on proliferation, migration and expressions of interleukin-8(IL-8), C-X-C motif chemokine-12(CXCL-12) and vascular endothelial growth factor(VEGF) in hepatoma HepG2 cells in vitro, in order to elucidate the anti-liver cancer activity of menthol and relevant mechanisms. Method: Different concentrations of menthol (25, 50, 100, 200 μmol·L-1) and blank group were applied to Hepatoma HepG2 cells. Cell counting kit-8(CCK-8) and EDU were used to detect the proliferation effect of menthol on HepG2 cell. Transwell experiment was used to detect the migration effect of menthol on HepG2 cell. Real-time fluorescent quantitative polymerase chain reaction technique was used to detect the inflammatory chemokines IL-8 and CXCL-12 mRNA expression levels in HepG2 cell. Western blot was used to detect the expression level of VEGF in HepG2 cells treated with menthol. Result: Compare with the blank group, menthol (25, 50, 100, 200 μmol·L-1) significantly inhibited proliferation and migration of HepG2 cells in vitro. When the concentration of menthol was 100 μmol·L-1 microns, the inhibitory effect was significant (P-1) significantly inhibited the expression levels of IL-8, CXCL-12 mRNA and VEGF protein in HepG2 cells (PConclusion: Menthol has an inhibitory effect on the proliferation and migration of hepatoma HepG2 cells in vitro, and the potential anti-liver cancer mechanism might be related to the inhibition of IL-8, CXCL-12 and VEGF expressions in the cells. This conclusion provides the experimental basis for elucidating the effect of menthol against liver cancer.

Hydrogen-water ameliorates radiation-induced gastrointestinal toxicity via MyD88’s effects on the gut microbiota

Although radiation therapy is a cornerstone of modern management of malignancies, various side effects are inevitably linked to abdominal and pelvic cancer after radiotherapy. Radiation-mediated gastrointestinal (GI) toxicity impairs the life quality of cancer survivors and even shortens their lifespan. Hydrogen has been shown to protect against tissue injuries caused by oxidative stress and excessive inflammation, but its effect on radiation-induced intestinal injury was previously unknown. In the present study, we found that oral gavage with hydrogen-water increased the survival rate and body weight of mice exposed to total abdominal irradiation (TAI); oral gavage with hydrogen-water was also associated with an improvement in GI tract function and the epithelial integrity of the small intestine. Mechanistically, microarray analysis revealed that hydrogen-water administration upregulated miR-1968-5p levels, thus resulting in parallel downregulation of MyD88 expression in the small intestine after TAI exposure. Additionally, high-throughput sequencing showed that hydrogen-water oral gavage resulted in retention of the TAI-shifted intestinal bacterial composition in mice. Collectively, our findings suggested that hydrogen-water might be used as a potential therapeutic to alleviate intestinal injury induced by radiotherapy for abdominal and pelvic cancer in preclinical settings.

Effect of HMGB1-siRNA on proliferation and apoptosis of HepG2 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of decreased expression of high mobility group Box-1 on the proliferation and apoptosis of HepG2 cells.</p><p><b>METHODS</b>Three specific siRNAs of HMGB1 were designed and synthesized, and were transiently transfected into HepG2 cells by Lipofectamine 2000. The HMGB1 expression in HepG2 cells was detected by RT-PCR and Western blotting respectively. The proliferation activity in vitro was assessed by MTT assay. In situ apoptosis was evaluated by terminal deoxynucleotidyl transferase-deoxyuridine triphosphate nick end labeling (TUNEL) assay.</p><p><b>RESULTS</b>All of these specific HMGB1-siRNAs (1, 2, 3) efficiently and specifically inhibited the expression of the HMGB1 gene, and the levels of HMGB1 mRNA were 1.147+/-0.024, 1.014+/-0.042, 0.435+/-0.055, respectively, in HMGB1-siRNAs transfection group, which were significantly lower than that in Lipofectamine 2000 alone group (1.411+/-0.065, P < 0.01). Correspondingly, all of these specific HMGB1-siRNAs (1, 2, 3) could efficiently and specifically inhibit the expression of the HMGB1 protein, and the levels of HMGB1 protein were 0.369+/-0.035, 0.340+/-0.028, 0.097+/-0.020, respectively, in HMGB1-siRNAs transfection group, which were significantly lower than that in Lipofectamine 2000 alone group (0.553+/-0.051, P < 0.01). Of the 3 specific HMGB1-siRNAs, HMGB1-siRNA-3 (siRNAH3) had the highest inhibition rate (80%). The proliferation of HepG2 cells was markedly inhibited by siRNAH3 transfection. Compared to mock-transfection, siRNAH3 transfection dramatically suppressed the proliferation of HepG2 cells (P < 0.01). Moreover, siRNAH3 can induce apoptosis (P < 0.01).</p><p><b>CONCLUSION</b>siRNA targeting HMGB1 mRNA can specifically reduce HMGB1 gene and protein expression. siRNAH3 can effectively suppress the proliferation and induce apoptosis of HepG2 cells.</p>