An Analysis On Pharmacognosy Of Iris Japonica And Its Antitumor Activity In Vitro Of Different Extracts Of Polar Solvent

Objective:Iris japonica Thunb.perennial,a type of Iris plant defined as Iridaceae,is widely distributed in China,which is commonly used as folk herb and landscaping plants,with the effect of decreasing swelling,relieving pain,rehabilitation,and detoxification.It is a promising material medicinal resource.The application of Iris japonica is mostly among folks,which often mixed with the Belamcanda chinensis and Iris tectorum materials;and the research dedicating in Iris japonica is rarely seen.On the ground of that,establishing a comprehensive and effective quality evaluation system can ensure the safe and effective use of drugs,and provide scientific basis for expanding the search for new drug sources.The pharmacognostic identification of Iris japonica was carried out in this experiment from the perspectives of character identification,microscopic identification,and physicochemical of Iris japonica,using DNA molecular marker technique establishing the molecular identification method and genetic diversity investigation.The antitumor activity of several tumor cell lines was screened by using different solvent extracts from Iris japonica Thunb,which provided a preliminary theoretical and experimental basis for the development of the medical resources of Iris japonica,the designation of quality standard and the research of antitumor substance base.Methods:firstly,the author applies source identification,character identification,microscopic identification,TLC identification,physicochemical identification to carry out the pharmacognosy study of Iris japonica.Next,conduct DNA molecular identification and genetic diversity study of the Iris japonica and its related species with the utilization of molecular marker of ISSR and SCoT.At the same time,in order to investigate the antitumor effect of the different solvent extractions of Iris japonica,this paper extracted the Iris japonica extractum with different-polar organic solvents;thereby the inhibition effects of five different solvent extraction constituents are conducted,i.e.cultured human hepatoma carcinoma cell lines Hep G2,human cervical carcinoma cell lines Hela,human lung adenocarcinoma cell A549,human colon carcinoma cell lines Hccc-9810,and human lung carcinoma cell lines H460.The extraction sites and the sensitive tumor cell lines with high inhibitory rate against tumor proliferation were screened,thereby providing a theoretical basis for the further development and utilization of Iris japonica resources.Results:1.Iris japonica Thunb.is a type of Iris plant defined as Iridaceae,which is a perennial herb with upright stem.The root and rhizome of the medicinal herbs present irregular,curved,and unbranched strips,with irregular longitudinal wrinkles,distinct,annular and elevated nodes,which have withered petiole or petiole residues.The root’s cross section has a large number of leaf-trace vascular bundle and fiber bundle;the middle column sheath is surrounded by many vascular bundles,and a large number of oxalic acid cubic and columnar crystals are found.There are stone cells found in the powder,which is dominated by stellate hair and mesh duct.Parenchyma cell wall presents irregular polyhedron.In physical and chemical experiments,flavonoids and triterpenes substances are found.TLC identification is carried out with chloroform-methanol-glacial acetic acid（10-2-0.2）as expansion agent,from which it is found that the separation degree of the rhizome extracts of Iris japonica is decent,wherein the positions corresponding to tectorigenin and apigenin have apparent spots.2.Molecular identification and genetic diversity studies were carried through on Iris japonica and its related species such as Belamcanda.And 10 ISSR primers and SCoT primers were screened out,which were extended to 130and 143 bands respectively,and the polymorphism bands were 96.2%and97.9%respectively.The Shannon Diversity Information Indexes（I）of the 12samples of materials were 0.5402 and 0.5003;Nei’s Gene Diversity Indexes（H）were 0.3630 and 0.3273 respectively,which indicated that the genetic difference between the 12 materials was instinct.The genetic distance obtained by SCoT marker analysis was higher than what obtained by ISSR marker analysis.The cluster analysis based on SCoT marker indicated that Iris japonica,Belamcanda chinensis,and Iris tectorum can be independently formed when the similarity coefficient is higher than 0.74;therefore they can be well distinguished from other materials of iris.3.In-vitro antitumor activity showed that the different extracts of polar solvent of Iris japonica had different proliferation inhibition effects on human hepatoma carcinoma cell lines Hep G2,human cervical carcinoma cell lines Hela,human lung adenocarcinoma cell A549,human colon carcinoma cell lines Hccc-9810,and human lung carcinoma cell lines H460.It was found that the extracts of ethyl acetate and N-butanol had better inhibitory activity on tumor growth,and showed a benign dose-effect relationship,in which the effect of ethyl acetate on proliferation of Hep G2 cells was most positive;IC₅₀is 53.08μg/ml.The inhibitory effect of N-butanol extracts on Hep G2 human hepatocellular carcinoma cell lines was relatively obvious,of which the IC₅₀was 174.91μg/ml,while water,petroleum ether extracts,and 95%ethanol extracts showed an insignificant inhibition effect,indicating that Iris japonica’s antitumor activity was mainly concentrated in ethyl acetate extract and N-butanol extract.Conclusion:This study identified the origin of Iris japonica and its relative species.ISSR and Scot markers can be applied stably,rapidly and accurately in the molecular identification of Belamcanda chinensis and Iris tectorum herbs.SCoT marker is superior to ISSR marker when it comes to the study of genetic diversity of Iridaceae population.The antitumor activity of different solvent sites of Iris japonica was screened preliminarily,and it was found that ethyl acetate and n-butyl alcohol showed decent activity,which laid a solid foundation for further study of the effective antitumor activity of Iris japonica,and provided a theoretical basis for the medicinal development of Iris japonica resources.