Effect of Dunhuang Pingweiwan and Its Decomposed Recipes on Tumor Inhibition and PI3K/Akt/mTOR Pathway in SCG-7901 Gastric Cancer Mice / 中国实验方剂学杂志

Objective:To investigate the antitumor effect and the mechanism of Dunhuang Pingweiwan and its decomposed recipes based on phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway in SCG-7901 gastric cancer-mice. Method:The subcutaneous tumor bearing model of SCG-7901 gastric cancer in mice was established， and the the mice were randomized into model group， Dunhuang Pingweiwan group （14.04 g·kg^-1·d^-1）， Huoxue Jiedu group （6.50 g·kg^-1·d^-1）， Wenzhong Sanhan group （3.64 g·kg^-1·d^-1） and cisplatin group （2 mg·kg^-1·d^-1）， with 8 mice in each group. From the 8^th day of inoculation， the mice were administered for 10 consecutive days. The mice were weighed and the general conditions were observed every other day. On the next day of the last administration， the mice were sacrificed， and the tumor was removed and weighed to calculate the anti-tumor rate. The histopathological changes were observed by hematoxylin-eosin （HE） staining， and the mRNA and protein expressions of PI3K， Akt， and mTOR in tumor tissues were detected by real-time polymerase chain reaction（Real-time PCR） and immunohistochemistry （IHC）， respectively. Result:From the 10^th day of inoculation， the mice in cisplatin group were generally in poor condition and their body mass decreased. The mice in model group， Dunhuang Pingweiwan group， Huoxue Jiedu group and Wenzhong Sanhan group were generally fair， and their body mass increased without significant difference among groups. The tumor inhibition rates of Dunhuang Pingweiwan， Huoxue Jiedu， Wenzhong Sanhan and cisplatin groups were 30.74%， 24.80%， 4.19% and 63.84%， respectively. Except for Wenzhong Sanhan group， tumor weight of the other treatment groups was significantly lower than that of the model group （<italic>P</italic><0.01）， and there was no significant difference between the Dunhuang Pingweiwan and Huoxue Jiedu group. Dunhuang Pingweiwan and Huoxue Jiedu group could significantly reduce tumor cell density and cause tumor cell necrosis. Compared with the model group， the expressions of PI3K， Akt， and mTOR mRNA and protein in the Dunhuang Pingweiwan， Huoxue Jiedu and cisplatin groups significantly decreased （<italic>P</italic><0.05， <italic>P</italic><0.01）， and there was no significant difference between the Dunhuang Pingweiwan group and Huoxue Jiedu group. Conclusion:Dunhuang Pingweiwan and its decomposed recipes （Huoxue Jiedu） have a certain anti-tumor effect on the SCG-7901 gastric cancer-mice， and the mechanism may be related to the down-regulation of key molecules in the PI3K/Akt/mTOR signaling pathway.

Protective Effect of Astragulus Polysaccharide on Chromosome Damage in Human BMSCs Exposed to Formaldehyde / 中国实验方剂学杂志

Objective::To study the protective effect of astragalus polysaccharide (APS) on micronucleus and sister chromatid exchange (SCE) in human bone marrow mesenchyml stem cell (BMSCs) exposed to formaldehyde, in order to initially explore the potential mechanism. Method::BMSCs were cultured in vitro, cells were randomly divided into five groups: control group, formaldehyde group, and APS 40, 100, 400 mg·L-1 groups. BMSCs were infected with 120 μmol·L-1 formaldehyde, meanwhile, APS 40, 100, 400 mg·L-1 groups were co-cultured with 40, 100, 400 mg·L-1 APS. Cell morphology was observed by inverted phase contrast microscope, micronucleus were detected by micronucleus test, SCE was detected by SCE test, and mRNA and protein expressions of proliferating cell nuclear antigen (PCNA), xeroderma pigmentosum B, D, F, G (XPB, XPD, XPF, XPG) were detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR)and Western blot. Result::Compared with control group, cell counts decreased, and cell morphology of BMSCs in formaldehyde group significantly changed, they were all recovered gradually in 40, 100, 400 mg·L-1 APS groups. Compared with control group, the micronucleus and SCE increased significantly (P<0.01), PCNA mRNA and protein expressions down-regulated significantly (P<0.05), while XPB, XPD, XPF, XPG mRNA and protein expressions up-regulated significantly (P<0.05, P<0.01). Compared with formaldehyde group, BMSCs were treated with APS at 40, 100, 400 mg·L-1, micronucleus and SCE decreased significantly (P<0.01), and mRNA and protein expressions of PCNA, XPB, XPD, XPF and XPG up-regulated significantly (P<0.05, P<0.01). Among them, the 100 mg·L-1 APS group had the most obvious effect. Conclusion::APS can protect formaldehyde-induced BMSCs micronucleus and SCE, especially 100 mg·L-1 APS has the most obvious effect. The mechanism may be associated with the up-regulation of expressions of PCNA, XPB, XPD, XPF and XPG in the nucleotide exicision repair pathway (NER), which promoted the damage repair.

Protective effect of Astragulus polysaccharide on DNA damage in human BM-MSCs exposed to formaldehyde / 中草药

Objective: To study the protective effect of Astragalus Polysaccharide (APS) on DNA damage in human BM-MSCs exposed to formaldehyde and to initially explore the potential mechanism. Methods: BM-MSCs were cultured in vitro and divided into control group, formaldehyde group, and APS at 40, 100, and 400 μg/mL groups. Proliferation activity was measured by MTT assay, DNA strand breakage was detected by comet assay, DNA-protein crosslinks (DPCs) was detected by KCl-SDS precipitation assay, and the mRNA and protein expression of XPA, XPC, ERCC1, RPA1 and RPA2 were detected by qRT-PCR and Western blotting. Results: Compared with model group, formaldehyde-contaminated BM-MSCs were treated with APS at 40, 100, and 400 μg/mL, the cell proliferation activity was increased significantly (P < 0.01), DNA strand breakage and DPCs level were decreased significantly (P < 0.01), and the mRNA and protein expression of XPA, XPC, ERCC1, RPA1, and RPA2 were up-regulated significantly (P < 0.05, 0.01). Among them, the effect of 100 μg/mL APS group was the most obvious. Conclusion: APS can protect formaldehyde-induced BM-MSCs DNA damage, especially 100 μg/mL APS has the most obvious effect. The mechanism may be associated with the up-regulation of XPA, XPC, ERCC1, RPA1, and RPA2, which promoted the repair of DNA damage.

Effects of Banxia Baizhu Tianma Decoction on Expression of miRNA-146a-5p in Hippocampus of Epileptic Rats and Bioinformatic Analysis / 中国中医药信息杂志

Objective To lay the foundation for studying the possible pathogenesis of epilepsy and the anti-epileptic mechanism of Banxia Baizhu Tianma Decoction through the bioinformatic analysis of target gene prediction and signal pathway of miRNA-146a-5p in hippocampus of epileptic rats. Methods Lithium-pilocarpine was used to induce seizures in rat models. The experiment rats were randomly divided into normal control group, model group, Banxia Baizhu Tianma Decoction group, with 20 rats in each group. The method of miRNA expression profiling was used to observe the miRNA differential expression of hippocampus neuron cell of rats. The expression level of miRNA-146a-5p was detected by real-time quantitative PCR. MiRDB was used for target gene prediction of miRNA-146a-5p, and miRTarBase and DAVID were used for enrichment analysis on the GO function and KEGG signaling pathway. Results The attack times and grades of the rats in Banxia Baizhu Tianma Decoction group were significantly lower than those in the model group from behavioral observation. MiRNA microarray analysis showed that the expression level of miRNA-146a-5p in model group was 2.107 times normal control group (P＜0.05), and the expression level decreased to 1.377 times after treatment with Banxia Baizhu Tianma Decoction (P＜0.05). The results of RT-PCR was consistent with that of miRNA microarray, with statistical significance (P＜0.05). MiRNA-146a-5p target gene prediction results had 140 target genes by GO, and there were 14 annotation information of biological process (P＜0.05), 9 annotation information of cellular component (P＜0.05), 11 annotation information of molecular function (P＜0.05). Enrichment analysis of KEGG biological pathway showed that 140 target genes of miRNA-146a-5p were enriched in EB virus infection signal pathway and thyroid hormone signaling pathway (P＜0.05). Conclusion miRNA-146a-5p is closely related to the inflammatory reaction after epilepsy, and Banxia Baizhu Tianma Decoction can control epilepsy possibly by controlling the inflammatory reaction after epilepsy.

The influence of the total flavonoids of Hedysarum polybotry on the proliferation, cell cycle, and expressions of p21Ras and proliferating cell nuclear antigen gene in erythroleukemia cell line K562 / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of total flavonoids of Hedysarum polybotry on the proliferation, cell cycle, and expressions of p21(Ras) and proliferating cell nuclear antigen (PCNA) gene in erythroleukemia cell line K562.</p><p><b>METHODS</b>The effect of total flavonoids of Hedysarum polybotry on K562 cell line survival was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) reduction assay. The time- and dose-dependent manner was also observed. The cell cycle and apoptosis were analyzed with flow cytometry (FCM). The immunocytochemistry method was applied to quantitatively analyze the effects of flavonoids of Hedysarum polybotry on changes p21(Ras) and PCNA gene expressions.</p><p><b>RESULTS</b>Flavonoids of Hedysarum polybotry (20-100 μg/mL) significantly inhibited the proliferation of K562 cells in a time- and dose-dependent manner. After K562 cells were cultured for 48 h, total flavonoids of Hedysarum polybotry had no significant effect on the apoptosis of K562 cells but showed significantly inhibition (P<0.01), indicating that total flavonoids of Hedysarum polybotry could induce K562 cells arrested at G(0)/G(1) and G(2)/M phases. Compared with the control group, p21(Ras) and PCNA gene expressions were decreased significantly in K562 cells treated with total flavonoids of Hedysarum polybotry (40 and 80 μg/mL, respectively) for 48 h.</p><p><b>CONCLUSION</b>The inhibitory effect on proliferation of K562 cells was observed in the groups treated with flavonoids of Hedysarum polybotry, which might be related to cells arresting.</p>