Study On The Chemical Constituents Of Conioselini Rhizoma Et Radix And Quality Evaluation Of "Gaoben" Herbs

The root and rhizome of Ligusticum sinense and Ligusticum jeholense (Umbelliferae) are used as a kind of TCM herbs, i.e. Ligustici Rhizoma et Radix, and the former is called as "Gaoben" and the latter as "Liao Gaoben" in China. It can relieve rheumatic pains, dispel cold, dehumidify moisture and relieve pain, thus it is usually used as treatment for cold, headache and pain in rheumatoid arthritis. Recently, natural resources of Ligustici Rhizoma et Radix became more and more exhausted because of environment destruction and overexploitation. The annual production of wild Ligustici Rhizoma et Radix reduced from5000t in2001to8001in2012. Although Conioselinum vaginatum belongs to genus Conioselinum of Umbelliferae, its rhizome and roots (Conioselini Rhizoma et Radix) are also used as the substitute of Ligustici Rhizoma et Radix, which is also called as "Xinjiang Gaoben" in China, mainly distribute in the regions of Tianshan and Altai mountains of Xinjiang Uyghur Autonomous Region. Rich natural resource and successful cultivation of this species in Ili Kazak Autonomous Prefecture of Xinjiang might provide a potential way to supply declining natrual resource of Ligustici Rhizoma et Radix. However, there are no overall scientific data to support Xinjiang Gaoben to substitute for Ligustici Rhizoma et Radix. We carry out the study on chemical constituents of Xinjiang Gaoben and quality evaluation of "Baoben" herbs (Xinjiang Gaoben, Gaoben and Liao Gaoben) to provide scientific evidences for the substitution of Xinjiang Gaoben for Ligustici Rhizoma et Radix.First, chemical constituents and bioactivities of Conioselinum vaginatum were summarized by referring to literatures and monographs. Then, we designed the schemes of market, cultivated and wild resource survey, and went to An’guo of Hebei province and Bozhou of Anhui province to survey market situation of "Baoben" herbs. And we went to Ili Prefecture of Xinjiang to survey cultivated and wild resource of Xinjiang Gaoben and collected experimental samples. We also made a comparative analysis of habit, distribution, output and market of "Baoben" herbs according to herbarium and literature data.We selected the Xinjiang Gaoben samples collected from Nilka county, Zhaosu county and Tekes county of Ili Kazak Autonomous Prefecture of Xinjiang, and conducted the study on the chemical constituents of its95%ethanol extracts, and sixteen compounds were isolated and identified from by silica gel column, Sephadex LH-20gel column and recrystallization. On the base of their physicochemical properties and spectroscopic data their structures were identified as Ligustilide (1),1,3-Dilinolein (2), Coniferaldehyde (3), Myristicin (4), Stigmasterol (5),β-sitosterol (6), Vanillin (7), Pregnenolone (8), Bergapten (9), Xanthotoxin (10), Methyl indole-3-carboxylate (11), Ferulic Acid (12),(E)-3-methoxy-4,5-methylenedioxcinnamic alcohol (13), p-hydroxybenzaldehyde (14),3-methoxy-4,5-methylenedioxy acetophenone (15), a-(ethoxymethyl)-4-hydroxy-Benzenemethanol (16). Among them, compound15was a new natural product, and compounds2,3,10,11,14,16were obtained from this genus for the first time.Secondly, the method for simultaneous determination of three compounds (ferulic acid, myristicin, ligustilide) and HPLC fingerprint of "Gaoben herbs" were established. The content of ferulic acid, myristicin and ligustilide in the18batches of Gaoben herbs were analyzed, the average content of ferulic acid in Gaoben, LiaoGaoben, and Xinjiang Gaoben was samilar, the average content of myristicin in Xinjiang Gaoben was obviously higher, and the average content of ligustilide in Gaoben and Liao Gaoben was quite higher. The HPLC fingerprint analysis of all the18samples was conducted, and there were some differences among Gaoben, LiaoGaoben, and Xinjiang Gaoben, but the same kind of Gaoben herbs collected from different origins showed good similarities. In addition, cluster analysis of the18samples was conducted by using SPSS software, the result showed that Xinjiang Gaoben with different origins were divided into one cluster, Gaoben and Liao Gaoben were divided into another cluster, the result were consistent with the results of Pharmacognosy identification.Finally, we compared the anti-inflammatory, analgesic and sedative bioactivity of Gaoben, Liao Gaoben and Xinjiang Gaoben. Xylene induced mouse auricle swelling model was adopted for anti-inflammatory activity research. Thermal stimulation induced pain model in mice was chosen to study their analgesic activity. And the model of increasing pentobarbital sodium-induced sleeping time was used to study their sedation activity. As a result, all the ethanol extracts of "Gaoben" herbs exhibited obvious inhibitory effect against the mice ear swelling induced by xylene with different intensity: Liao Gaoben> Gaoben> Xinjiang Gaoben. Liao Gaoben and Gaoben could remarkably relieve the pain induced by heat in mice, while Xinjiang Gaoben showed no obvious inhibitory effect. All the ethanol extracts of "Gaoben" herbs could inhibit the central nervous system in cooperation with pentobarbital sodium with different intensity: Xinjiang Gaoben> Liao Gaoben> Gaoben.In conclusion, the resources status and chemical constituents of Xinjiang Gaoben were studied in this theme, and it is helpful to develop and utilize this plant resources reasonably. Meanwhile, the quality evaluation of Gaoben herbs was carried out, and the results showed there were some differences between chemical constituents and biological activities among Xinjiang Gaoben, Gaoben and Liao Gaoben. So it is necessary to study further to get scientific evidences of Xinjiang Gaoben as the substitutes of Ligustici Rhizoma et Radix.