Studies Of Activity Evaluation And Exploiture Of Functional Product On TCM Mao-Ren-Shen

Mao-Ren-Shen (Cat Ginseng) is one of the most frequently used traditional Chinese medicine (TCM) for antitumor therapy, especially against alimentary canal cancers in East China. In order to empolder TCM resources in China, the following studies were carried out.1. Confirmation of genuine resources and biological characteristics of Mao-Ren-ShenA. macrosperma was proved to be the original source of Mao-Ren-Shen, while the recorded resource A. valvata may be the minor resource at most, according to the results of folk and market survey, identification of plant and raw material and history of medicinal application and scientific name. In addition, the pharmacognosy characters of Mao-Ren-Shen were obtained by the study of histology features and microscopic characters, which contributed new evidence to the distinction of this plant. The truebom species was discriminated from adulterants for sale, typically A.valvata which was similar in shape and used to be recognized as the original source of Mao-Ren-Shen, with the methods of morphology of wild plant and raw material, crystalloid morphology, traditional identification and modern analytical technics, such as fluorescent reaction, UV spectrum, fluorescence spectrum and so on.2. Studies on phytochemistryChemical components in Mao-Ren-Shen were extracted and separated by the technique of solvent-refining. Extractions successively extracted with petroleum ether, chloroform, ethyl acetate, butyl alcohol and water, and then a phytochemical screening was proceeded. Using the method of brine shrimp-tumor cell activity tracking and bioassay-guided filtration, we made a system isolation study on the chemical constituents. From the active chloroform fraction, we gained several triterpenoids. The most active ones among all the triterpenoids were identified as 2a,3β,19a,23-tetrahydroxyursa-12-en-28-oic acid (II₃), 2a,3a-dihydr-oxyurs-12,18-dien-28-oic acid (II₅), 2a-hydroxyoleanolic acid (II₉) and 2a,3β,23-trihydroxyursa-12-en-28-oic acid (II₁₆), which were obtained from this plant for the first time. Moreover, it was investigated that the molecular mechanism of II₅-induced cytotoxicity against A549 was characterized with the induction of apoptosis and the influence on cell cycle.3. Study on pharmacologyWe investigated the pharmacological effect of Mao-Ren-Shen on antitumour, immunomodulatory and antibacterial activities with multiguide multitarget in vivo and vitro. According to the result of MTT assay, A549 was the most sensitivity one of the selected 12 cell lines of human or mouse cancer. The fractions of petroleum ether (IC₅₀=262.90μg/mI), chloroform (IC₅₀=39.04μg/ml) and ethyl acetate (IC₅₀=75.22μg/ml) were also found effective. In vivo antitumor test, the AM fraction (petroleum ether, chloroform and ethyl acetate incorporate fractions) of Mao-Ren-Shen exhibited pronounced activity against inoculated tumor S180 and A549 with a dose-dependent manner. Mao-Ren-Shen showed the potent antioxidant activity based on DPPH, reducing power and TBA tests in a dose dependent manner, which can broaden its therapeutic applications towards the prevention of degenerative and neoplastic diseases of various organs. In addition, the results for the test in vivo showed its aqueous extract significantly increased the overall immune functions on sarcoma 180 bearing mice, whereas pronounced increased immune functions on natural mice. Finally, we analysed the antibacterial activity of the plant. The data demonstrated a mild activity in its essential oil, and the test fungi were more sensitive than the bacterial. Moreover, a high linalool content (48.17%) seemed to be responsible for the bacteriostatic and fungistatic properties of the essential oil.4. Exploration and applicationAccording to the function and indication of the formula and the properties of Mao-Ren-Shen, as well as its pharmacology function, the route of preparative procedure and index components were ascertained. We investigated the optimal adjuvant in tablet prescription, abstraction technology, prepration method, quality and quantity standard, initial stabilization and the pharmacological measuration. The results of research showed that this functional product technology was reasonable and feasible, the quality was stable and reliable, and the product could modulate the immune functions.