Effect of differentiation of cardiac stem cells mediated by pilose antler polypeptides on expressions of terminal myocardial differentiation genes ANP and MLC-2v / 吉林大学学报(医学版)

Objective:To explore the effect of differentiation of cardiac stem cells(CSC)mediated by pilose antler polypeptides on the expressions of terminal myocardial differentiation genes atrial natriuretic polypeptide (ANP)and myosin light chain 2v(MLC-2v),and to clarify the mechanism of repairing the damaged myocardium.Methods:The healthy male Wistar rats born 2 d were selected to extract the CSC.The surface marker c-Kit and the purity of CSC were identified by immunofluorescence and flow cytometry.The culture dish was used as the experimental unit and the extracted cells were divided into the following four groups:blank control group(the same amount of buffer was added for induction),5-azacytidine group(induced with 3 μmol·L-15-azacytidine),pilose antler polypeptides group(induced with 800 mg·L-1pilose antler polypeptides)and combined group(induced with 800 mg·L-1pilose antler polypeptides and 3 μmol·L-15-azacytidine);the cells were incubated for 48 h in the condition of 37℃ and 5% CO2.The expression levels of ANP and MLC-2v in supernatant of the cells in various groups were detected by ELISA method.The expression levels of ANP and MLC-2v mRNA in the cells in various groups were detected by RT-PCR method.Results:CSC were prepared with the purity>95%.The results of ELISA showed that the expression levels of ANP and MLC-2v in 5-azacytidine,pilose antler polypeptides and combined groups were significantly increased compared with blank control group(P<0.05).The expression levels of ANP and MLC-2v in combined group were increased compared 5-azacytidine and pilose antler polypeptides groups,but there were no significant differences(P>0.05).The result of RT-PCR showed that the expression levels of ANP and MLC-2v mRNA in 5-azacytidine,pilose antler polypeptides and combined groups were significantly increased compared with blank control group(P<0.05).Conclusion:Pilose antler polypeptides can induce the differentiation of CSC into cardiac cells by promoting the expressions of ANP and MLC-2v,and they can reduce the cytotoxicity induced by 5-azacytidine in some degrees.

Effect of pilose antler polypeptides on apoptosis and membrane stability of mitochondria in cardiac stem cells / 中国组织工程研究

BACKGROUND: Cardiac stem cells can differentiate into cardiomyocytes in vitro under induction of pilose antlerpolypeptides, which provides a new therapeutic idea for myocardial injury.OBJECTIVE: To explore the effect of pilose antler polypeptides on the apoptosis rate and membrane stability ofmitochondria in cardiac stem cells.METHODS: We chose healthy male Wistar rats born 2 days to extract cardiac stem cells. The culture dish was used asthe experimental unit, and extracted cells were divided into the following four groups (n=12). Blank control group: Thesame amount of buffer was added for induction; 5-azacytidine group: induced with 5-azacytidine (3 μmol/L); pilose antlerpolypeptides group: induced with pilose antler polypeptides (800 mg/L); combined group: induced with pilose antlerpolypeptides (800 mg/L) and 5-azacytidine (3 μmol/L). After 48 hours induction, apoptosis rate of cardiac stem cells ineach group was detected with flow cytometry. The membrane potential of mitochondria in cardiac stem cells wasdetected with immunofluorescence. The expression level of Nkx 2.5, GATA4, ATF-2, and MEF-2C in cardiac stem cellswas detected using western blot assay.RESULTS AND CONCLUSION: There were small, round and bright cells after 2 weeks culture, and cell colonies ofcardiac stem cells formed. The apoptosis rate of cardiac stem cells in the 5-azacytidine group increased significantly (P 0.05). The expression level of MEF-2C in each group was at a middle level, and there were no significantdifferences among groups (P > 0.05). To conclude, our experimental findings indicate that pilose antler polypeptidescould decrease the apoptosis rate and improve membrane stability of mitochondria in cardiac stem cells. Themechanism may be related to the increased expression of Nkx 2.5, but whether the mechanism is related to GATA4,ATF-2 and MEF-2C needs to be further studied.

Prevention and therapeutic effects of sika deer velvet collagen hydrolysate on osteoporosis in rats by retinoic acid / 中国中药杂志

The objective was to evaluate the preventive and therapeutic effects of the collagen hydrolysate extracted from Sika deer velvet (CSDV) on osteoporosis rats induced by retinoicacid. Histomorphometric indices and serum biochemical parameters were measured in osteoporosis rats treated with/without antler collagen and in sham-operated rats. Our results were as follows: compared with the osteoporosis group, significant elevation in the levels of bone mineral density (BMD), Ca, P and static histomorphometric indexes and biomechanical properties, but reduction in the level of serum alkaline phosphatase (ALP) were observed in antler collagen-treated groups. However, the above function with the collagenase solution velvet material varied with the different doses. In conclusion, the extracted collagen is found to play a role in the prevention and treatment of osteoporosis rats by retinoic acid.

Preparation of recombinant human zona pellucida 3 protein and its immunologic activity / 中国免疫学杂志

Objective:To prepare and purify recombinant human zona pellucida 3 protein,and to study its immunologic activity.Methods:The E.coli BL21 containing recombinant plasmid pGEX4T-1/ huZP3 was induced to express GST-fusion protein by IPTG.After a series of purification procedure,the purified protein was analyzed by SDS-PAGE.After the mice immunized with rhuZP3,the antibody responses against rhuZP3 were detected by ELISA.Results:The soluble fusion protein was expressed,and purity of rhuZP3 was 95%.Moreover, purity of rhuZP3 could be recognized by anti-human ZP3 in ELISA.Conclusion:The rhuZP3 is obtained through the preparation of prokaryotic expression system and anti-rhuZP3 antibody has immunological activity.

Experimental research on cardiotonic effect of active compound from raw and processing aconite roots / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the cardiotonic effect of active compoud from raw and processing aconite roots used rats with acute heart failure (AHF) as the experimental animals.</p><p><b>METHOD</b>Isolated toad cordis was employed to observe the cardiotonic effect and the AHF models were reproduced in rats by pentobarbital sodium to observe the hemodynamics marker acute heart failure, and elucidate cardiotonic effect.</p><p><b>RESULT</b>The DDA(s) and MDA-P improved contractile force obviously on isolated toad cordis (P < 0.05). And the most amplification of contractile force was 91.107% +/- 87.663% (P < 0.01). But the effect on isolated heart rates was not significant. The change of LVSP, +/- dp/dt(max), LVEDP were significant (P < 0.01, P < 0.001), and the HR increased evidently by adminstrate processing aconite roots active compounds after the agent in 5 min.</p><p><b>CONCLUSION</b>There are cardiotonic effect on isolated toad cordis in rats with active compounds from raw and processing aconite roots.</p>

On anti-inflammation,immune regulation and anti-stress effect of cartialgenous collagen zymolyte / 中华中医药杂志

Objective:To investigate the anti-inflammation,immune regulation and anti-stress effect of cartialgenous collagen zymolyte.Methods:The anti-inflammatory effect was observed by carrageenan-induced foot swelling test,the immune effect was observed by delayed-type hypersensitivity (DTH) induced by 2,4-dinitrochlorobenzene and the anti-stress effect was observed by hypoxia tolerance and fatigue tolerance. Results:The cartialgenous collagen zymolyte can inhibit the carrageenan-induced foot swelling and delayed-type hypersensitivity (DTH) induced by 2,4-dinitrochlorobenzene,and improve the macrophagocyte phagocytosis function,and enhance the body,s non-specific resistance to fatigue.Conclusion:Cartialgenous collagen zymolyte appeared obvious immune regulation,anti-inflammation and anti-stress effect.

Protective Effect of Fuzheng Chuyi Granules on the Mice Infected with Influenza Virus / 中药新药与临床药理

Objective To observe the influence of Fuzheng Chuyi Granules on the mice infected with influenza virus and to observe its antibiotic and antivirus actions in vitro.Methods The death rate of mice infected with influenza virus in each group was compared and antimicrobial activity of Fuzheng Chuyi Granules was observed in vitro.Hep-2 passage cells were subcultured to observe the protective effect of the maximal non-toxic dosage(TD0),the infective dose of 50% tissue culture(TCID50) and other different dosage of Fuzheng Chuyi Granules on cells infected virus.Results Compared with that in negative control group,the death rate of infected mice was lower in Fuzheng Chuyi Granules groups.Fuzheng Chuyi Granules had antibiotic action on bacteria in vitro,particularly on streptococcus pneumoniae.Meanwhile,Fuzheng Chuyi Granules could alleviate cytopathy induced by virus.Conclusion Fuzheng Chuyi Granules have a certain protective effect on the animal infected with virus,and also have antibiotic and antivirus actions.