The Identification For The Separation And The Effectiveness Of Dendrocandin Ingredients

Dendrobium, one of the biggest plant in the orchidaceae grow mainlyin the warm and humid southern environment, the further north, the lessspecies it would be. There are more than70kinds of Dendrobium growingin China, including Dendrobium officinale, Dendrobii, ring grassDendrobium, Dendrobium nobile whip Dendrobium and so on.. Dendrobiumofficinale, ranking the first among the “nine mesona”, recorded inthe “Pharmacopoeia of the People’s Republic of China “(2005) is one ofthe three Dendrobium that can be taken as medical use. The Dendrobiumofficinale is mainly distributed in Yunnan, Zhejiang, Guangxi and Hunanand some other places. The main component of Dendrobium officinale areDendrobium polysaccharide, alkaloid, stilbene compounds, amino acids andtrace elements. The polysaccharides content of Dendrobium officinale isrelatively high, while the alkaloid content is low. Some researchers likeGuo Shun-Xing has extracted18new compounds from Dendrobium officinalesince2008, and named it Dendrobium officinale A-R which has a varietyof medicinal effects, including anti-tumor, anti-inflammatory,anti-radiation, regulating the function of the immune, reducing bloodglucose and cholesterol, and so.This topic applies the column chromatography, reverse phase columnchromatography and thin layer chromatography method and a higher levelsof an ethanol-soluble component has been extracted from Dendrobiumofficinale in Pu’er City, Yunnan Province. The component appearing asshallow brown amorphous solid, is sent to the CAS Kunming Institute ofBotany for identification, where the component is first found in Dendrobium officinale and named as Dendrocandin S.We have researched the anti-inflammatory effect by using themacrophages differentiated from the Dendrocandin S functioned on THP-1cell Model of inflammatory cells. We have used the Dendrocandin S to handlemacrophage a half hour, to cause the macrophage inflammatory reaction withlipopolysaccharide (LPS), to take the cell culture supernatantrespectively in3hours,6hours and9hours using the ELISA method todetect the level of inflammatory factors cell secretion. From the results,we found that Dendrocandin S has significant inhibitory effect to thesecretion of the cytokine TNF-a, compared with the positive control group,when the dense of the Dendrocandin S is25μg/ml, but the inhibitoryeffect is not significant when the dense is12.5μg/ml and6.25μg/mlshowing that the Dendrocandin S has the function of preventing andsuppressing the inflammation with dense dependency in inhibition.Inspired by the phenomenon of the Dendrocandin S’ inhibition onTNF-α secretion, we made a preliminary mechanism study on suppressinflammation Dendrocandin S.30minutes after Dendrocandin S acted onthe macrophages, with LPS to induce inflammation, then we extractedsamples of total RNA from the cells after reverse transcriptase RealTimePCR method to detect changes in cellular mRNA expression, respectivelyin0.5hour,1.5hours,2.5hours and3.5hours. From the results, we foundthat Dendrocandin S has the obvious inhibition the macrophage TNF-α andIL-6mRNA when the dense is25μg/ml compared with the positive controlgroup, but not obvious to IL-1βmRNA, showing the affect difference ofDendrocandin S between IL-1β, TNF-α and IL-6.30minutes afterDendrocandin S acted on the macrophages, with LPS to induce inflammation,we extracted the cells protein respectively in0.5,1and1.5hours,and detected,by using the Western Blot method, the changes of NF-κBphosphoric acid. We learned from the results, compared with positive control, the Dendrocandin S has obvious inhibition to phosphorylation ofNF-κB. Because NF-κB activation can cause inflammation and tumor, sothe inhibition of NF-κB phosphorylation may inhibit inflammation andtumorigenesis.When we use Dendrocandin S to handle the apoptotic cell-Jurkat cellswith the dense of50μg/ml and25μg/ml in order to observe the changesof cell morphology, We found that after the Dendrocandin S acted on thecells for half a hour, the cells have signs of cell of turning around,and the pseudopodia disappear with the control group compared to the moreobvious changes in cell shape morphogenesis. When the cells had beenhandled by Dendrocandin S for24hours, with the acted dense of50μg/ml,the Jurkat cells almost all died, and the death rate decreases to50%within the acted dense of25μg/ml, and the state of the survival cellsare not good showing that Dendrocandin S has the function of causing thecells death, and dense dependency.Jurkat cell morphology observation experiments reveal that, theDendrocandin S can cause the changes of the Jurkat cell morphology. Whatare the causes? We use the Western Blot method to detect early changesin histone acetylation in cell morphology associated protein α-Tubulin,in order to make clear the early changes of Tubulin protein. We found thatwhen the Dendrobium hormone concentration is25μg/ml, and the handlingprocess of Jurkat cells for half an hour, the cells of α-Tubulinacetylation level of shows significantly increasing with the negativecontrol group, this results indicate that the Dendrocandin S can increasethe stability of the Jurkat cells, α-Tubulin protein when its dense is25μg/ml, which might be an important mechanism for promoting apoptosis.We also used flow cytometry to detect the death caused by theDendrocandin S to Jurkat cells, to find that Dendrocandin S can cause deathto it and has dense dependency. we want to know whether the death caused by cell necrosis or by apoptosis, we detected with Western Blot theapoptosis marker proteins of PARP and found that the PARP protein has beencut when Jurkat cells are handled by the Dendrocandin S with the denseof25μg/ml for2hours, indicating that this promoting death is causedby the apoptosis. Are these apoptosis caused through p53pathway? Wedetected with Western Blot Jurkat, the cells p53increasing apoptosis tocontrol protein PUMA and found that the Dendrocandin S with theconcentration of25μg/ml can decrease PUMA protein of Jurkat cells,indicating that the pro-apoptotic effect caused by Dendrocandin S within25μg/ml to Jurkat cells is not dominated by the p53pathway.Dendrobium officinale, one of the important kind herbs in theTraditional Chinese Medicine have a high use value, and more and moreattention has been paid to Dendrobium officinal. This essay has done someresearches on both the efficacy of the initial mechanism after extractingfrom Dendrobium officinale, a new component of Dendrocandin S, and itsanti-inflammatory and anti-tumor. The study will provide some datasupporting Dendrobium officinale to be widely liked and accepted andprovide the impetus for the development of Dendrobium officinale industry,to make it well known both at home and abroad.