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Chrysanthemum effective part of the role of the brain Liver Screening_7533

By Jason Reyes,2014-10-30 14:00
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Chrysanthemum effective part of the role of the brain Liver Screening_7533

    Chrysanthemum effective part of the role of the brain Liver Screening

     Author: Ren Ai-nong, LIU Chun-yu, Yang Nian-yun

     Abstract Objective To daisy brain extracts with different polarity (CFDN) in the effective screened Protective effect of liver position. Methods 0.1? L4 (10ml/kg) ip in mice, resulting in acute liver injury model, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), hepatic tissue lesions; with CCl4 injury in rat liver cells, MTT Determination of liver cell proliferation. Results CFDN 60% ethanol and 95% ethanol elution column parts can significantly reduce CCl4 liver injury in mice serum ALT and AST activity (P <0.01, P <0.05), pathological observations indicate that: The site could significantly reduce the liver the extent of injury (P <0.01, P <0.05), CFDN 60% ethanol and 95% ethanol elution column positions can significantly increase activity of liver cells, promote liver cell proliferation (P <0.01). Conclusion CFDN 60% ethanol and 95% ethanol column elution position has significant

    hepatoprotective effect.

     Key Words chrysanthemum brain; effective parts; CCl4; liver injury

     Screening of the active fractions from Chrysanthemum nankingense on their liver-

    protective effects

     Abstract Objective To screen the active fractions from Dendranthema

    Nankingense of Chrysanthemum nankingense on their liver-protective effects.Methods

    The mouse models of carbon tetrachloride-induced acute hepatic injury were employed

    in this study.The activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum were assaied.Also pathological changes were observed in liver tissues.The proliferous activities were assaied by MTT in cells of liver-

    injured rat by 0.1? l4 (10ml/kg) ip.Results The CFDN of 60% alchohol and 95% alchohol can debase ALT and AST activities in the serum of liver-injuried mice (P

    <0.01, P <0.05). This can elevate the activities of liver cells and accelerate hepatocellular proliferation.The pathological observation indicated the degrees of inflammatory activity in liver tissues were markedly reduced (P <0.01). Conclusion The results show that CFDN of 60% alchohol and 95% alchohol have highly antihepatotoxic effect.

     Key words Chrysanthemum nankingense; effective site; carbon tetrachloride;

hepatic injury

     Department of Chrysanthemum morifolium Ramat brain brain (Chrysanthemum nankingense Hand-Mezz) Spear of the dry leaves, Jiangsu real estate resources, and its bitter, Xin, sexual cool; functions: Qingrejiedu. The variety in 2002 in Jiangsu Province Science and Technology Department Fund for Nature project, after study, to extract from this plant were isolated 12 compounds, mainly 26 acid, sitosterol, Kim Sung-grass

    Flavin, luteolin, etc. in which flavonoid luteolin is a representation of constituents [1]. Parts of different polarity of the plant by in vivo antibacterial, anti-inflammatory, anti-

    virus tests are indicative of the corresponding active site. On this basis, we conducted a study of its hepatoprotective effect. Using 0.1? L4 (10ml/kg) ip in mice, resulting in

    acute liver injury model, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), hepatic tissue lesions, as well as with CCl4 injury in rat liver cells, MTT method Determination of liver cell proliferation was observed with different

    polarity chrysanthemum brain parts (CFDN) the role of the Liver.

     1 Materials

     1.1 Drug Chrysanthemum brain collected in Nanjing Laoshan, the original plant by the Nanjing University of Chinese Medicine Professor Liu Xunhong brain identified as

    chrysanthemum (Chrysanthemum nankingense Hand-Mezz), take the brain

    chrysanthemum leaves 80% ethanol extract, extract was extracted by petroleum ether extracts A , liquor on the macroporous resin column with 20% ethanol, respectively, 60% ethanol, 95% ethanol elution, may extract B, C, D (total flavonoids contents were 10%, 50%, 25%). Temporary use during saline or dimethyl sulfoxide dissolved. DDB tablets (Jiangsu Peng Harrier Pharmaceutical Co., Ltd.), the Pro with normal saline, when crushed into a fine powder made from suspension.

     1.2 Reagents CCl4 analysis of alcohol (Nanjing Chemical Reagent Factory), to dilute the olive oil into a 0.1% solution; 4 yl methyl chloride salt, even (MTT, AMERSO subpackage); alanine aminotransferase (ALT), aspartate aminotransferase (AST)

    determination of reagent box (Shanghai coxim Biotechnology Research Institute); sodium dodecyl sulfate (AMRESCO-packing); RPMI1640 (Gico); newborn calf serum

    (Hangzhou Institute of Sijiqing biological materials); rat liver cells (Cote d'Ivoire Cell

    Biology Institute).

     1.3 Animals Kunming mice, male, SPF grade, 16 ~ 20g, by the Experimental Animal Center of Soochow University.

     2 Methods

     2.1 CCl4 mouse model [2] and drug treated mice were randomly grouped as normal

    group, injury model group, DDB control group (30mg/kg), CFDN (A), CFDN (B), CFDN (C ), CFDN (D) group, CFDN group grouped 200mg/kg, 400mg/kg two-dose

    group. Ig normal group and model group, normal saline, the other groups of drugs ig corresponding day a second consecutive 6 days. 7 days after administration, in addition to the normal group were given an equal volume of olive oil, the remaining mice in each group ip 0.1% CCl4 0.1ml/10g, fasting, 16h after the removal of the eye to take blood, preparation of serum, serum ALT and AST activity; take small pieces of left lobe of liver buy 10% formalin fixed for hematoxylin - eosin staining of liver tissue changes

    observed under light microscope. Reposted elsewhere in the paper for free download

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     2.2 Determination of cell proliferation MTT method with reference Yue-Hua Li, etc.

    [3] the determination of liver cell suspension regulates cell number of 5 × 104/ml, plus

    cell suspension in 96-well plates, 100μl / hole, according to the different concentrations of added drugs and their sub - groups, each with 6 complex holes. Normal group,

    adding RPMI-1640 culture medium; the other groups to join CCl4 solution to a final

    concentration of 10mmol / L, the test group also added to the corresponding drug, 24h after the culture plate by adding 5mg/ml MTT solution, each well 20μl, incubated by adding after 4h 10% sodium dodecyl sulfate dissolved in 100μl suspension response and

    a black crystal, place 12h after the microplate reader was measured at 570nm wavelength absorbance (A) values, reflecting the proliferative activity.

     2.3 statistical methods data (x ? s) that it had adopted SPPS10.0 statistical

    procedures dealing with single-factor analysis of variance between the two groups using Dunnett method of comparison, the level of information with Ridit test.

     3 Results

     3.1 pairs of CCl4 liver injury in mice the effects of serum ALT and AST compared

    with the normal group, model group mice ALT and AST increased significantly (P <0.01); Compared with model group, CFDN 60% ethanol elution position of high-dose

    group and CFDN 95% ethanol elution positions of high-dose group of mice serum ALT

    and AST decreased significantly (P <0.01, P <0.05), prompted CFDN 60%, 95% ethanol elution positions of CCl4 liver injury in mice serum ALT and AST activity were significantly increased antagonism. Table 1.

     3.2 pairs of CCl4 liver injury in mice the impact of pathological changes in liver

    tissue pathology light microscope observations, model group, liver cell injury serious, a bit like necrosis, cloudy swelling, ballooning hepatocytes, portal area shows severe inflammatory cell infiltration, cytoplasm filled with large eosinophilic coarse granules, severe focal necrosis can be seen. CFDN 60% ethanol and CFDN 95% ethanol column elution positions of high-dose group of mice significantly reduced liver cell damage, we can see a few spotty necrosis of liver cells, and eosinophilic changes of liver cells, other groups had no significant improvement of liver injury in mice. Table 2. Table 1 CFDN pairs of CCl4 liver injury in mice serum ALT and AST activity Note: Compared with model group, P <0.05, P <0.01 Table 2 CFDN on CCl4 liver histopathological changes of note: with the normal group, ΔP <0.01; Compared with model group, P <0.05, P <0.01

     3.3 CCl4 injury in rat liver cell proliferation activity as compared with the normal group, model group, rat liver cell proliferation activity was significantly lower (P <0.01), CFDN 60%, 95% ethanol column elution fractions of dose group, large dose group compared with model group, liver cell proliferation activity was significantly increased (P <0.01), its role was a definite dose-effect relationship. Table 3. Table 3

    CFDN pairs of CCl4 injury in cultured rat liver cell activity Note: with the CCl4 group, P <0.01; with the DMSO group, P <0.01

     4 Discussion

4.1 CCl4-induced liver injury liver injury model is the standard model of drug

efficacy of lipid peroxidation is the main mechanism of liver injury. CCl4 into the body

through the hepatic P450 enzymes activate the drug trichloromethyl peroxy radical (

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