| PART1：Caenorhabditis elegans (referring as the C.elegans in the followingpresentation) is a category of non-parasitic C.elegans living in the soil, which isused to be the first multicellular eukaryotes to clarify the sequence of the entiregenome. Due to the short lifespan, the transparent body, and the easiness to cultivateand observe, the C.elegans becomes an excellent model organism in the field of lifescience research. In recent years, many experimental groups applied the C.elegans toestablish the longevity model and conducted the pharmacological research, becausein the process of cultivation, the C.elegans will continue to consume material addedin the medium, maintain the concentration of drug in its body, and the changes in theorganizational structure and behavior expression of the C.elegan from developmentto death can be observed under the microscope, making it possible to conduct asystematic research on the biological phenotype. Therefore, the C.elegans becomesthe appropriate model organism in the rasearching of the biological function of theactive constituent in traditional Chinese herb.In the first part of this article, we prepare a medium contained certainconcentration of the active constituent in traditional Chinese herbs, then feed themwith the different medium in the administration group and the control group, observeand compare the life span in each situation and record other biological phenotype.We conclude that all the Betaine with the concentration of100μg/ml and300μg/ml,the Ginseng polysaccharide with the concentration of40μg/ml and200μg/ml, andthe conversation of Dendrobium water-soluble components into crude drug with theconcentration of0.67g/ml and0.067g/ml can post an effect of extending the lifespan of the C.elegans, while the Notoginsenoside and “Crab legs” post none.Daphnetin with the concentration of300μg/ml and500μg/ml will shorten the lifespan and leading to dysplasia. All these provide the theoretical basis for the development and utilization of the related the active constituent in traditionalChinese herbs.The second part of the paper reveals the effect of the daphnetin on thebiological function of the C.elegans. There are two aspects that the daphnetin canaffect the the biological function of the C.elegans: shorten the life span and developewith dysplasia. From the experimental results, we conclude that the shorten effect oflife span brought by the daphnetin on the C.elegans is similar to thecumulative toxic and genomics effect, and this life shortening phenomenon isincreasing with the rising of concentration. In addition, the relations between theabnormal development of the C.elegans and the cumulative toxic and genomicseffect may be very weak, because the proportion of dysplasia of the F2is lower thanthe F1. These results provide a toxicological basis and foundation for thedevelopment and utilization of the daphnetin drugs.In the third part of this article, we conduct a research on the resistance effect toUV radiation of the pine nut oil microencapsulated powder through theestablishment of a model of UV damage. The pine nut oil microencapsulated powderwith the concentration of50μg/ml and100μg/ml presents the resistance effect on thelow level of UV radiation with100J/m2, while this protection disappears on the levelof the UV radiation rising to800J/m2. The life span of the C.elegans can beextended on the appropriate concentration of pine nut oil microencapsulated powder(50μg/ml,100μg/ml), which provides a theoretical basis for the development anduse of the products of the pine nut oil microencapsulated powder UV radiation.Although a lot of pharmacological effects of the active constituent in thetraditional Chinese Herb have been proven in the human body and a variety ofanimal models, none of the effects on the C.elegans life activities have been reportedsystematically. As a higher eukaryotic multicellular organism, the C.elegans is veryconducive to our life research due to its short life span, and of great value to conductthe pharmacology research due to its physiological characteristics. Therefore, moreand more experimental groups use the C.elegans to research the biological functionof the traditional Chinese Herb active constituent. In this research, we takeadvantage of the C.elegans model, conduct a systematic analysis of the biologicalfunction of the active constituent of Chinese herbal medicine, that provide a new theoretical basis for the research and development of the active constituent of theseherbs. PART2：Diabetic mellitus (DM) is recognized as a group of metabolic disordersand characterized by hyperglycemia and dyslipidaemia. Type1diabetes ischaracterized by a lack of insulin production and type2diabetes results from thebody’s ineffective use of insulin, Insulin resistance in its metabolic target tissues.Type3diabetes is gestational diabetes. Type2diabetes mellitus (T2DM) is the mostcommon form and accounts for around90%of all diabetes worldwide. Insulinbinding evokes a cascade of phosphorylation events, beginning with theautophosphorylation of the IR on multiple tyrosyl residues. Autophosphorylationenhances IR kinase activity and leads to recruitment of insulin receptor substrate(IRS) proteins, followed by activation of phosphatidylinositol3-kinase (PI3K) anddownstream protein kinase AKT, subsequent translocation of the glucose transporterGLUT4and regulation of glucose uptake. Therefore, Improving tissue sensitivity toinsulin is a major clinical goal to help ameliorate abnormal metabolism.Protein tyrosine phosphatases (PTPs) constitute a large and structurally diversefamily of signaling enzymes that control the cellular levels of protein tyrosinephosphorylation and play an important role in the intracellular signal transductionprocess and metabolism. SHP-1(SH2-containing tyrosine phosphatase1), also calledHCP, SHPTP1and PTP1C, is an important member of protein tyrosine phosphotasesubfamily with highly conserved sequence. SHP-1is a kind of cytosolic proteincontaining two SH2domains at the N-terminal and one catalytic domain at theC-terminal. It was reported that increased SHP-1activity led to immuno-deficientsyndrome andn leukemia. The most recent studies have shown that SHP-1is relatedto type II diabetes and high level of blood sugar is due to elevated SHP-1activitywhich means that endogenous SHP-1plays a critical role in regulating blood sugarmetabolism and SHP-1may be a potential target for reducing the blood sugar of diabetes patients by inhibiting its activity.Two steps of ion-exchange chromatography, Q-Sepharose Fast Flow andSP-Sephadex, were used to isolate and purify SHP-1protein obtained from E.coliDE3strain. The purity of target protein was more than90%determined bySDS-PAGE and HPLC analysis. The enzyme showed a linear relationship during thefirst20min. The catalytic domain of SHP-1protein tyrosine phosphatase, is moresensitive towards the Perilla stem extract than other PTPs and its IC50was4μg/ml.Based on the above experimental phenomenon, we investigated thehypoglycemic mechanism of Perilla stem. The tyrosine phosphorylation levels ofthe IR β-subunit and ERK1/2significantly increased in the HepG2cells by theconcentration of1mg/ml and0.75mg/ml of Perilla stem compared to the controlgroup.We speculate that the hypoglycemic mechanisms of Perilla stem may becaused by inhibition of the SHP-1activity, consequently, activating the signalingpathway, which also provide us a theoretical basis for Perilla stem on type2diabetictherapy.40mice were selected for establishing type2diabete animal model which wereinjected with STZ/kg per day. After the treatment, each mouse was weighed everyday and the blood sugar was detected every week using Blood Glucose Kit. Themouse with more than11.1mmol/L blood sugar was considered as established model.The mice were randomly grouped into control group, herb-treated group, and modelgroup. Both the control group (healthy mice) and the negative control group (type IIdiabetic mice) were administrated with25ml/kg water according to body weights,while the herb-treated group (type II diabetic mice) was administrated with10g/kgherbs according to body weights for25days. Our animal experiments providedstrong evidence that perilla stem significantly reduced the blood sugar of type2diabetes animal model with19.4mmol/L.The Perilla stem has hypoglycemic effectthrough inhibiting the protein phosphorylation of SHP-1, which may lead to thesecondary development of Chinese traditianal herbs. |
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