哈薩克藥阿爾泰瑞香不同提取物對癌細(xì)胞增殖的抑制作用(英文)課件

1、Antiproliferative activity of different extracts from Daphne altaica Pall. on selected cancer cellsProf. Dr. Murat KizaibekTraditional Kazakh Medicine Research Institute of Ili Kazakh Autonomous Prefecture, P. R. ChinaAlmaty May 3, 2012IntroductionCancer is the worlds second biggest killer after car

2、diovascular disease. In 2005, 7.6 million people died from cancer, more than HIV/AIDS, malaria and tuberculosis combined. In addition, this number is expected to rise to 9 million in 2015 and increase further to 11.5 million in 2030 (WHO, 2007). Clearly, there is a great need to improve current canc

3、er therapies and to search for new therapies.Data from Statistics Canada Throughout history, plants have afforded a rich source of compounds that have found many applications in the fields of medicine, pharmacy and biology. Within the sphere of cancer, plants have played an important role as a sourc

4、e of effective anti-cancer agents, and it is significant that over 60% of currently used anti-cancer agents are derived in one way or another from natural sources, including plants, marine organisms and micro-organisms (Cragg et al., 2005).Daphne altaica Pall., locally known as uwsoyq or qasqr jiyde

5、k, is a deciduous herb of the Thymelaeaceae family. It is endemic to the north of Jungar Basin of Xinjiang, China (the Tacheng and Habahe areas), Altai, Manrak and Tarbagatai Mountains of Kazakhstan and Altai region of Russia as well as northwest MongoliaD. altaica have long been used in TKM to trea

6、t esophagus cancer, gastric cancer, tracheitis, common cold, soar throat, rheumatism, snakebite, and for their antitussive and diaphoretic properties (Xu et al., 2009). Its medicinal use was firstly recorded in a Kazakh medical classic Shipagerlik Bayan, which was written by Oteyboydak Tleukabyluly

7、in 15th century (Tleukabyluly, 1994). Oteyboydak Tleukabyluly（1388-？）Shipagerlik BayanTranslation：Prescription 4573 : add the bark of Daphne into the meat broth and boil them. As soon as the broth comes to a full boil, remove the residue, drink the broth and eat the meat. It can cure asthma and coug

8、h.At page 420 of this book, there is the following prescription ：Although many benefits of this plant have been claimed, no scientific datum is available so far about its biological properties, especially its anticancer activity. Therefore, this prompted us to investigate the antiproliferative activ

9、ity of this plant on cancer cells. For these purposes, six extracts were prepared and tested for their potential antiproliferative properties with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) bioassay on different cancer cell lines. To our knowledge, this is the first time that

10、 the anticancer activity of D. altaica is evaluated.Plant materialThe barks of D. altaica were collected from Altai Mountain, Xinjiang, China in July 2008. The plant was identified and authenticated by Bahargul Kongirkhan, a herbalist at the herbarium of Altay Institute for Drug Control, Xinjiang, C

11、hina. A voucher specimen (No. 050036) was deposited at the same place.Tested material Dried barks of the plant (150 g) were chopped and extracted with 95% EtOH by maceration for 2 weeks in a dark place at a room temperature of 20 2. The procedure was repeated for twice. The extracts were combined, c

12、oncentrated under reduced pressure and freeze-dried to yield the EtOH extract (DA-Et, 12.10 g ). 0.5 g of this extract was kept for MTT assay and the rest was submitted to a sequential liquid-liquid extraction with solvents of increased polarity to yield petroleum ether (DA-Pt，1.7064 g), chloroform

13、(DA-Ch,0.6915 g), ethyl acetate (DA-Ea，1.8237 g), n-butanol (DA-Bu，2.5242 g) and aqueous (DA-Aq，2.5998 g) fractions.Cell proliferation assayAll the samples were tested at 6.25, 12.5，25，50，100 g/ml concentrations. The samples were dissolved in DMSO and further diluted with cell culture medium. Cells

14、incubated with the same concentration of DMSO were used as a control. The DMSO final concentration was adjusted to 1% of the total volume of medium in all treatments, including the control. For MTT assay, 1 105 cells/well were plated into 96-well plates (Nunclon, Denmark) and incubated for 24 h befo

15、re the addition of drugs. After 48 h of incubation for all cells, 20 l of MTT (Sigma, USA) reagent ( 5 mg/ml) in phosphate buffered saline (PBS) was added to each well. The plates were incubated at 37C for 4 h. At the end of the incubation period, the medium was removed and pure DMSO (150 l) was add

16、ed to each well. The metabolized MTT product was quantified by reading the absorbance at 490 nm on a Beckman Coulter-AD340 (Beckman Coulter, Fullerton, CA, USA). Results were expressed percentage of cell viability (%). All assays were performed in triplicate. The median growth inhibitory concentrati

17、on values (IC50) were used to compare the antiproliferative activity of extracts on cancer cells.Statistical analysisThe results of percentage of cell viability were presented as means SD. Statistical comparisons between treatment groups and the control group were performed using the one-way ANOVA f

18、ollowed by post hoc Tukey (in case of equal variance) or Dunnetts T3 (in case of unequal variance) test. These tests were performed using SPSS 13 for Windows. p 0.05 was considered statistically significant. IC50 values and their 95% confidence interval (IC95) were calculated using sigmoidal dose-response model with variable slo