Research Update:Chinese Pulsatilla Root.Pulsatilla chinensis.

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   Phytochemical info of Chinese Pulsatilla Root.Pulsatilla chinensis.

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 Definition:Chinese Pulsatilla Root.Pulsatilla chinensis are majorly composed of
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   Research Update:Chinese Pulsatilla Root.Pulsatilla chinensis.

  Preparation of 23-hydroxybetulinic acid from the roots of Pulsatilla chinensis.:Zhong Yao Cai. 2007 Feb;30(2):170-2. Chinese.Zhao HN, Wang Y, Su WK, Zhang XQ, Ye WC.Institute of Traditional Chinese Medicine & Natural Products, Jinan University, Guangzhou 510632, China.

 OBJECTIVE: To isolate and purify high purity 23-hydroxybetulinic acid from the roots of Pulsatilla chinensis. METHODS: By acid hydrolysis, silica gel column chromatographies and recrystalization methods, 23-hydroxybetulinic acid was obtained from the roots of P. chinensis. RESULTS: High purity 23-hydroxylbetuline acid was isolated from the roots of P. chinensis, the contents is over 98. 0%. CONCLUSION: The preparation process is feasible and convenient. High purity 23-hyroxybetulinic acid could be used in the quality control of the materials and medicament of 23-hydroxybetulinic acid.

  Effect of anemonin on NO, ET-1 and ICAM-1 production in rat intestinal microvascular endothelial cells.:J Ethnopharmacol. 2006 Apr 6;104(3):362-6. Epub 2005 Oct 27.Duan H, Zhang Y, Xu J, Qiao J, Suo Z, Hu G, Mu X.Department of Animal Science and Technology, Beijing Agricultural College, Beijing 102206, PR China.

 Anemonin (the dilactone of cyclobutane-1, 2-diol-1, 2-diacrylic acid) was isolated from the root of Pulsatilla chinensis Regel. Pulsatilla chinensis Regel has been used in the treatment of enteritis in China for years. However, only little was known about the mechanism underlying its anti-inflammatory effects. We investigated the effect of anemonin on the release of nitric oxide (NO), endothelin-1 (ET-1) and soluble intercellular adhesion molecule-1 (sICAM-1) induced by lipopolysaccharide (LPS) in primary cultures of rat intestinal microvascular endothelial cells (RIMECs). RIMECs were challenged with 1 microg/ml LPS with or without the presence of various concentrations of anemonin (1, 5 and 10 microg/ml). Anemonin significantly inhibited the production of NO and ET-1 induced by LPS at a concentration of 5 microg/ml and at 10 microg/ml anemonin down-regulated LPS-induced sICAM-1 expression. Anemonin itself had no effect on either factor. These findings suggest that anemonin may exert some beneficial therapeutic action in intestinal inflammation, at least in part by inhibiting the production of NO, ET-1 and ICAM-1 in RIMECs and thus preventing intestinal microvascular dysfunction.

  Microscopic identification of homonym drugs of "Baitouweng" by digital imaging technique.:Yao Xue Xue Bao. 2004 Oct;39(10):797-802. Chinese.Wang SY, Zhang M, Wang ZT.Department of Pharmacognosy, China Pharmaceutical University, Nanjing 210038, China.

 AIM: To identify the commercial drugs collected from 11 different areas with name of "Baitouweng", in order to understand the homonym status of Baitouweng in markets. METHODS: Based on macroscopic identification, we further studied the microscopic structures of the collected samples by digital imaging technique. RESULTS: Nine species belong to 4 different families have been found out from the commercial drugs of "Baitouweng". There are the roots of Pulsatilla chinensis (Bunge) Regel (recorded in Chinese Pharmacopoeia with name "Baitouweng"), P. cernua (Thunb.) Bercht et Opiz, P. turczaninovii Kryl. et Serg., P. dahurica (Fisch.) Spreng., Anemone tomentosa (Maxim.) Pei, Rhaponticum uniflorum (L.) DC. and the herbs of Potentilla chinensis Ser., Po. discolor Beg. and Polycarpaea corymbosa Lam.. CONCLUSION: The original plants of the crude drug "Baitouweng" were still promiscuous in the market because there are different medicinal usages in different areas resulting in the phenomenon of homonym for Baitouweng. Otherwise, the digital photographs offered by the paper visually reflected the main microscopic characteristics of the commercial "Baitouweng", can be used for the identification of the above drugs.

  Apoptotic activity of betulinic acid derivatives on murine melanoma B16 cell line.:Eur J Pharmacol. 2004 Sep 13;498(1-3):71-8.Liu WK, Ho JC, Cheung FW, Liu BP, Ye WC, Che CT.Department of Anatomy, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, People's Republic of China. ken-liu@cuhk.edu.hk

 The mitochondrion plays a crucial role in the process of apoptosis and has thus become one of the targets for the search for potential chemotherapeutic agents. Betulinic acid [3beta-hydroxy-lup-20(19)lupaen-28-carbonic acid], a lupane-type triterpene which is abundant in many plant species, has been shown to exert a direct effect on the mitochondria and subsequent apoptosis in melanoma cells. Chemical synthesis and modification of betulinic acid are being explored to develop more potent derivatives. We present here the apoptotic activity of several natural derivatives of betulinic acid which were isolated from the roots of a Chinese medicinal herb, Pulsatilla chinensis (Bge) Regel [Ye, W., Ji, N.N., Zhao, S.X., Liu, J.H., Ye, T., McKervey, M.A., Stevenson, P., 1996. Triterpenoids from Pulsatilla chinensis. Phytochemistry 42, 799-802]. Of the five compounds tested, 3-oxo-23-hydroxybetulinic acid was the most cytotoxic on murine melanoma B16 cells (IC50=22.5 microg/ml), followed by 23-hydroxybetulinic acid and betulinic acid (IC50=32 and 76 microg/ml, respectively), with lupeol and betulin exhibiting the weakest cytotoxicity (IC50> or =100 microg/ml). Exposure of B16 cells to betulinic acid, 23-hydroxybetulinic acid and 3-oxo-23-hydroxybetulinic acid caused a rapid increase in reactive oxidative species production and a concomitant dissipation of mitochondrial membrane potential in a dose- and time-dependent manner, which resulted in cell apoptosis, as demonstrated by fluorescence microscopy, gel electrophoresis and flow-cytometric analysis. Cell cycle analysis further demonstrated that both 3-oxo-23-hydroxybetulinic acid and 23-hydroxybetulinic acid dramatically increased DNA fragmentation at the expense of G1 cells at doses as low as 12.5 and 25 microg/ml, respectively, thereby showing their potent apoptotic properties. Our results showed that hydroxylation at the C3 position of betulinic acid is likely to enhance the apoptotic activity of betulinic acid derivatives (23-hydroxybetulinic acid and 3-oxo-23-hydroxybetulinic acid) on murine melanoma B16 cells.

  Chemiluminescence detection of superoxide anion release and superoxide dismutase activity: modulation effect of Pulsatilla chinensis.:Anal Bioanal Chem. 2004 May;379(1):171-7. Epub 2004 Feb 25.Yao D, Vlessidis AG, Gou Y, Zhou X, Zhou Y, Evmiridis NP.Laboratory of Analytical Chemistry, Department of Chemistry, University of Ioannina, 451 10, Ioannina, Greece.

 A novel flow-injection chemiluminescence-based method has been developed for determination of superoxide dismutase (SOD) activity. An in-vitro superoxide anion generation xanthine/xanthine oxidase stable source was established on line with FIA/CL-detection apparatus, for measuring SOD activity. This method can detect SOD in the linear range of 0.002-2.00 U mL(-1) with a detection limit of 0.001 U mL(-1). Another method for detection of superoxide anion is based on the luminol-FeCl(3) chemiluminescence (CL) reaction. This method was used to evaluate superoxide release and SOD activity in rats treated with the traditional Chinese herb Pulsatilla chinensis, which resulted in high clearance of hepatitis B virus (HBV) after treatment of a hepatitis B patient. Interestingly, we found that treatment with Pulsatilla chinensis can specifically increase superoxide release by liver tissues and, at the same time, slightly increase extracellular SOD (ECSOD) activity in plasma; in particular it can markedly increase MnSOD activity in mitochondria in liver tissue. This work revealed a possible mechanism whereby Pulsatilla chinensis prevents possible infection (for example HBV) by specifically increasing superoxide release in the liver and increasing MnSOD activity to minimize superoxide-mediated toxicity.


  New lupane glycosides from Pulsatilla chinensis.:Planta Med. 2002 Feb;68(2):183-6.Ye W, Zhang Q, Hsiao WW, Zhao S, Che CT.

 Two new lupane glycosides along with five known triterpenoids were isolated from the roots of Pulsatilla chinensis (Ranunculaceae). The structures of the new glycosides were determined to be 3-O-beta-D-glucopyranosyl(1-->3)-alpha-L-arabinopyranosyl-23-hydroxybetulinic acid 28-O-alpha-L-rhamnopyranosyl(1-->4)-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranosyl ester (pulsatilloside D, 6) and 3-O-[beta-D-glucopyranosyl(1-->4)][alpha-L-rhamnopyranosyl(1-->2)]-alpha-L-arabinopyranosyl-23-hydroxybetulinic acid 28-O-alpha-L-rhamnopyranosyl(1-->4)-beta-D-glucopyranosyl(1-->6)-beta-D-glucopyranosyl ester (pulsatilloside E, 7) by spectroscopic analysis and chemical methods. The compounds were evaluated for cytotoxic activities against K-562 human leukemia and HeLa cells.

  New bisdesmosidic triterpene saponins from the roots of Pulsatilla chinensis.:J Nat Prod. 2001 Sep;64(9):1226-9.

 Further phytochemical analysis aimed at the triterpene saponin constituents of the roots of Pulsatilla chinensis has resulted in the isolation of four new bisdesmosidic triterpene saponins whose aglycons are based on the lupane skeleton (1-4), together with three known saponins (5-7). The structures of the new compounds were determined by spectroscopic analysis and acid-catalyzed hydrolysis.

  Triterpene saponins and lignans from the roots of Pulsatilla chinensis and their cytotoxic activity against HL-60 cells.:J Nat Prod. 1999 Sep;62(9):1279-83.

 Two new triterpene saponins (8 and 9) and seven previously reported triterpene saponins (1-7) based upon oleanolic acid or hederagenin, along with two known lignans, (+)-pinoresinol (10) and beta-peltatin (11), were isolated from a saponin fraction prepared from the MeOH extract of the roots of Pulsatilla chinensis. The structures of the new saponins were determined by spectroscopic analysis, including 2D NMR spectroscopic techniques, and the results of hydrolytic cleavage. The isolated compounds and some derivatives were evaluated for their cytotoxic activity against HL-60 human leukemia cells.

  Triterpenoids from Pulsatilla chinensis.:Phytochemistry. 1996 Jun;42(3):799-802.Ye WC, Ji NN, Zhao SX, Liu JH, Ye T, McKervey MA, Stevenson P.Department of Phytochemistry, China Pharmaceutical University, Nanjing, China.

 A new lupane type triterpenic acid, pulsatillic acid, and two new lupane type triterpenoid glycosides, pulsatilloside A and B, along with the known 23-hydroxybetulinic acid were isolated from the roots of Pulsatilla chinensis. Their structures were characterized as 3-oxo-23-hydroxy-lup-20(29)-en-28-oic acid, 3 beta, 23-dihydroxy-lup-20(29)-en-28-oic acid 3-O-alpha-L-arabinopyranoside and 3 beta, 23-dihydroxy-lup-20(29)-en-28-oic acid 28-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside on the basis of hydrolysis and spectral evidence including two-dimensional relay HOHAHA, one-dimensional multiple relay COSY and ROESY NMR techniques. Pulsatillic acid exhibited cytotoxic activities against P-388, Lewis lung carcinoma and human large-cell lung carcinoma.

  Determination of ranunculin in Pulsatilla chinensis and synthetic ranunculin by reversed phase HPLC.:Yao Xue Xue Bao. 1990;25(12):932-5.Zhang XQ, Liu AR, Xu LX.Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing.

 A method for the determination of ranunculin in Pulsatilla chinensis and synthetic ranunculin by reversed phase HPLC using betamethasone as internal standard is described. The column employed was a 10 micron LiChrosorb RP-18 (230 x 14 mm), the eluting solvent consisted of methanol-water (40:10, V/V) and the effluent was monitored at 225 nm. The advantages of this method are simple, rapid and accurate. The coefficients of variation are less than 1%.


  Pulsatilloside C from the roots of Pulsatilla chinensis.:J Nat Prod. 1998 May;61(5):658-9.Ye W, He A, Zhao S, Che CT.Department of Phytochemistry, China Pharmaceutical University, Nanjing 210009, China.

 A new lupane-type triterpene saponin, pulsatilloside C (1), was isolated from the roots of Pulsatilla chinensis. Its structure was established to be 3beta,23-dihydroxylup-20(29)-en-28-oic acid 28-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)- beta-D-glucopyranoside.

  Assessment of Anemone pulsatilla for some biological activities:Pak J Pharm Sci. 1998 Jan;11(1):47-53.Saify ZS, Noor F, Mushtaq N, Dar A.Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Karachi, Karachi, Pakistan.

 Preliminary screening of herbal extracts employs the determination of their biological activity and the estimation of their therapeutic potential. This eliminates the unnecessary effort of fractionation and purification of constituents unimportant from the practice pharmacological point of view. It also gives a scientific basis to the herbs traditionally used in folk medicine. Thus, during the present investigation the spasmolytic effect of Anemone pulsatilla extract on isolated tissues of rabbit jejunum was evaluated. In addition the pharmacological screening of Anemone pulsatilla extract was also earned out.

  Structural rearrangements, including parallel inversions, within the chloroplast genome of Anemone and related genera.:J Mol Evol. 1994 Mar;38(3):274-81.Hoot SB, Palmer JD.University of Michigan Herbarium, Ann Arbor, MI 48109.

 Chloroplast DNA cleavage sites for 10 restriction enzymes were mapped for 46 species representing all sections of Anemone, four closely related genera (Clematis, Pulsatilla, Hepatica, and Knowltonia), and three more distantly related outgroups (Caltha, Ranunculus, and Adonis). Comparison of the maps revealed that the chloroplast genomes of Anemone and related genera have sustained an unusual number and variety of rearrangements. A single inversion of a 42-kb segment was found in the large single-copy region of Adonis aestivalis. Two types of rearrangements were found in the chloroplast genome of Clematis, Anemone, Pulsatilla, Hepatica, and Knowltonia: An approximately 4-kb expansion of the inverted repeat and four inversions within the large single-copy region. These rearrangements support the monophyletic status of these genera, clearly separating them from Caltha, Ranunculus, and Adonis. Two further inversions were found in two Clematis species and three Anemone species. While appearing to support a monophyletic grouping for these taxa, these two inversions conflict with data from both chloroplast restriction sites and morphology and are better interpreted as having occurred twice independently. These are the first two documented cases of homoplastic inversions in chloroplast DNA. Finally, the second intron of the chloroplast rps 12 gene was shown to have been lost in the common ancestor of the same three Anemone species that feature the two homoplastic inversions.

  Influence of permeation enhancers on transdermal permeation of anemonin.:Zhongguo Zhong Yao Za Zhi. 2007 Mar;32(5):393-6. Chinese.Ning YM, Rao YF, Liang WQ.School of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310031, China.

 OBJECTIVE: To investigate the effect of different permeation enhancer on transdermal permeation of anemonin through human skin. METHOD: The permeation experiments were performed using human skin on modified Franz diffusion cells in vitro. The concentrations of anemonin in receptor compartment at specified time points were determined by HPLC. The steady flux and the cumulative quantity of anemonin through skin were calculated. Result: The flux of anemonin permeating through human skin from 30% ethanol, 50% ethanol solution and a combination of 3% laurocapm -5% polysorbate 20 and 30% ethanol -3 % laurocapm -5% polysorbate 20 of anemonin was (9.30 +/- 0.32), (18.56+/-0.58), (7.29+/-0.35), (13.77+/-0. 16) microg x cm(-2) x h(-1) and 7.9, 15.9, 6.2, 11.8 times higher than from saturated water solution respectively. CONCLUSION: Ethanol and laurocapm can remarkably improve the transdermal permeation of anemonin and the anemonin have the potential to be developed to new transdermal preparation.


  Scientific References:

  1.Research Update:Chinese Pulsatilla Root.Pulsatilla chinensis.