Combination of Total Astragalus Extract and Total Panax Notoginseng Saponins Strengthened the Protective Effects on Brain Damage through Improving Energy Metabolism and Inhibiting Apoptosis after Cerebral Ischemia-Reperfusion in Mice

Objective： To explore the effects and molecular mechanisms of the combination between total Astragalus extract （TAE） and total Panax notoginseng saponins （TPNS） against cerebral ischemia- reperfusion injury. Methods： C57BL/6 mice were randomly divided into sham-operated group, model group, TAE （110 mg/kg） group, TPNS （115 mg/kg） group, TAE-TPNS combination group and Edaravone （4 mg/kg） group, treated for 4 days, then, cerebral ischemia-repeffusion injury was established by bilateral common carotid artery （CCA） ligation for 20 min followed by reperfusion for 1 and 24 h. Results： TPNS could increase adenosine triphosphate （ATP） level, TAE and TAE-TPNS combination increased ATP, adenosine diphosphate （ADP） contents and Na＋-K＋-ATPase activity, and the effects of TAE-TPNS combination were stronger than those of TAE or TPNS alone after reperfusion for 1 h. After reperfusion for 24 h, TAE, TPNS and TAE-TPNS combination significantly increased neurocyte survival rate and decreased the apoptosis rate as well as down-regulated the expression of phosphorylated c-June N-terminal kinasel/2 （p-JNK1/2）, cytochrome C （Cyt C）, cysteine aspartic acid-specific protease （Caspase）-9 and Caspase-3. Furthermore, the effects in TAE-TPNS combination were better than those in TAE or TPNS alone. Conclusion： The combination of TAE 110 mg/kg and TPNS 115 mg/kg could strengthen protective effects on cerebral ischemia injury, the mechanism underlying might be related to improving jointly the early energy metabolism, and relieving the delayed apoptosis via inhibiting the mitochondrial apoptosis pathway of JNK signal transduction.

The Effects of Astragalus Membranaceus Extract on Sperm Quality in vitro

This study is to explore the effects on sperm quality and fertilizing ability by using astragalus membranaceus extract as additive among infertile males. Human sperms were cultured in the concentrated astragalus membranaceus extract for 1 h, then semen parameters were measured by the equipment of computer assisted semen analyzer (CASA). The results are as follows: compared with the original semen, the sperm motility and velocity, the percentage of grade A sperms and the swaying frequency of sperm heads were improved markedly in the semen treated with astragalus membranaceus extract (P<0.05). Meanwhile, the astragalus membranaceus aqueous extract was analyzed by atomic absorption spectrophotometer. It was found there exist several metal elements and a certain amount of trace elements in the extract (10 mg/ml). The concentration of K +, Ca 2+ , Zn 2+ were 25.1 μg/ml, 7.28 μg/ml, and 0.47 μg/ml respectively. The influence of this additive on artificial insemination was also discussed. We considered that the studied Chinese herbal medicine additive has good effects on sperm preparation in vitro, and this might be a potential method in assisted reproductive technology.

Molecular Mechanism of Induction on Apoptosis of Human Esophageal Cancer HCE-4 Cells by Active Components from Astragalus membranaceus

[Objectives] To investigate the pharmacologic effects of active components from A. membranaceus on human esophageal cancer HCE-4 cells and its apoptosis mechanism. [Methods] The viabilities of HCE-4 cells were measured by MTT assay. The induction of active components from A. membranaceus on apoptosis of HCE-4 cells was detected by Annexin V-FITC/PI double staining. The apoptotic-related protein expression levels were determined by Western blotting. [Results] Formononetin and astragaloside IV suppressed the proliferation of HCE-4 cells in a dose-dependent manner. The Annexin V-FITC/PI double staining results showed that formononetin and astragaloside IV could induce HCE-4 cells apoptosis in a time-dependent manner. The Western blotting results showed that formononetin and astragaloside IV could significantly down-regulate p-AKT,pro-caspase-3,and increase cle-caspase-3 protein expression in HCE-4 cells. [Conclusions]Active components from A. membranaceus such as formononetin and astragaloside IV significantly inhibited the proliferation of human esophageal cancer HCE-4 cells by inducing mitochondrial dependent apoptosis via AKT signaling pathway.

Research on the quality control technology for the Astragalus membranaceus extraction solution in cosmetics

Calycosin-7-glucoside,which is the target ingredient of the Astragalus membranaceus extraction solution,was studied by the high-performance liquid chromatography-diode array detector method(HPLC-DAD),and a high-performance liquid chromatography-electrospray ionization detector method(HPLC-CAD)was also established for fingerprint chromatogram analysis.Meanwhile,the heavy metal content was tested to build a quality control standard for Astragalus membranaceus extraction solution.Result shows a good linearity in 4-80μg/mL mass concentration range with the correlation coefficient r=0.999,RSD=2.4%,and the recovery rate is between 99.2%-102.7%.The fingerprint chromatogram analysis features with 14 specific peaks,including 5 identified components.The similarity is between 0.485 to 0.995.Content of heavy metals such as lead,mercury,and cadmium are below the detection limit,and the content of arsenic is less than 2 mg/kg.This research results can serve as process optimization basis and quality control standards for Astragalus membranaceus extraction solution,and provide reference to quality control and research guideline for other cosmetics using herbal extracts as raw materials.