ABSTRACT
OBJECTIVE： To study the anti-tumor effect of ethanol extract of Smilax trinervula and its different polar extract parts， and to provide reference for the screening of anti-tumor active parts. METHODS： MTT method was used to detect the inhibitory rates of different concentrations of ethanol extract of S. trinervula， its petroleum ether， ethyl acetate， n-butanol and water layer extract parts to the proliferation of human hepatocellular carcinoma cell line HepG2， human lung cancer cell line A549， human breast cancer cell line MCF-7 and MDA-MB-231， human cervical cancer cell line HeLa and human ovarian cancer cell line HO-8910. Semi-inhibitory concentration （IC50） was calculated. A total of 80 KM mice were subcutaneously inoculated with S180 cell suspension in right forelimb axilla to induce tumor-bearing mice model. The mice were randomly divided into 8 groups： e.g. model group （normal saline， twice a day， i.g.）， cyclophosphamide （positive drug） group （0.025 g/kg， once a day， i.p.）， S. trinervula ethanol extract high-dose， medium-dose and low-dose groups， ethyl acetate part of ethanol extract high-dose， medium-dose and low-dose groups （0.1， 0.05， 0.025 g/kg by extract， twice a day， i.g.）， with 10 rats in each group. The mice in each group were given medicine for consecutive 14 days. The inhibition rate， spleen index and thymus index of mice in each group were measured after fasting for 12 hours after the last administration. RESULTS： In vitro experiments showed that S. trinervula and different polar extracts inhibited the proliferation of 6 kinds of tumor cells in significant dose-effect manner， especially ethyl acetate part of ethanol extract. IC50 of it to tumor cells was 40-210 μg/mL， that of it to MCF-7， MDA-MB-231 and HeLa cells were 70.56， 83.58， 44.67 μg/mL， respectively. In vivo experiments showed that the tumor mass of mice were decreased significantly in each administrations （P＜0.05 or P＜0.01）， the tumor mass of mice in S. trinervula ethanol extract high-dose group， ethyl acetate part of ethanol extract high-dose and medium dose groups were significantly lower than cyclophosphamide group （P＜0.01）. The spleen index of mice was decreased significantly in cyclophosphamide group， S. trinervula ethanol extract high-dose group， ethyl acetate part of ethanol extract high-dose and medium dose groups， compared with model group （P＜0.01）； thymus index of mice in cyclophosphamide group was increased significantly， compared with model group and other administration groups （P＜0.01）. CONCLUSIONS： The ethyl acetate part of ethanol extract of S. trinervula has the best anti-tumor effect and less immunosuppressive effect.
ABSTRACT
OBJECTIVE:To investigate the anti-inflammatory and analgesic effects of mongolian medicine Cymbaria dahurica extract. METHODS:96 KM mice(or SD rats)were randomly divided into model group(water),positive control group(aspirin, 0.5 g/kg),C. dahurica ethanol extract (70% ethanol) low-dose,medium-dose and high-dose groups (0.325,0.650,1.300 g/kg, calculated by crude drug) and C. dahurica aqueous extract of the residue of alcohol extraction low-dose, medium-dose and high-dose groups(0.325,0.650,1.300 g/kg,calculated by crude drug). They were given relevant medicine intragastrically,once a day,for consecutive 7 d. The xylene-induced ear edema method was used to determine the degree of ear edema,and egg white-in-duced paw edema method was used to determine paw edema after inducing inflammation 1,2,4,6 h;anti-inflammatory activity of C. dahurica extract was investigated. 96 KM mice were grouped and given medicine with same method;the number of writhing within 20 min was determined by acetic acid writhing method. Another 64 KM mice were grouped with same method,with 8 mice in each group;except positive control group was given tramadol hydrochloride(0.5 g/kg)intragastrically,other groups were given relevant medicine with same method. Pain thresholds of mice were determined by hot-plate test before and after medication 30,45, 60,90 min,and analgesic effects of C. dahurica extract were investigated. RESULTS:Compared with model group,C. dahurica extract could obviously restrain the ear edema of mice and paw edema of rats 6 h after egg white-induced inflammation. Except paw edema of rats in C. dahurica aqueous extract of the residue of alcohol extraction high-dose group was decreased slightly,there was statistical significance among other groups (P<0.05 or P<0.01). C. dahurica extract decreased the number of writhing in mice within 20 min,and extended pain thresholds of mice 30,60,90 min after medication (P<0.05). CONCLUSIONS:Both C. dahurica ethanol extract and aqueous extract of the residue of alcohol extraction posses certain anti-inflammatory and analgesic effects.
ABSTRACT
OBJECTIVE To observe the effect of natrin from Naja naja atra(Chinese cobra)on intracellular free calcium overload,and to discuss the protective effect and the possible mechanism of natrin on myocardium calcium(Ca2+)and potassium(K+)ion channels in the primary cardiomyocytes of SD neonatal rats. METHODS The primary cardiomyocytes of SD neonatal rats were used,which were respectively pretreated with natrin 5,25 and 125 mg · L-1 for 24 h before injury was induced by H2O2 0.3 mmol · L- 1. The dynamic variation of intracellular calcium was monitored by laser confocal microscopy using Fluo-3 as Ca2+fluorescence probe. Additionally,the cardio myocytes of neonatal rats were pretreated for 24 h using different concentrations of natrin 5,25,125 mg · L-1 and verapamil 5 nmol · L-1,followed by exposure to H2O2 0.3 mmol · L-1 for 15 min. Then,the mRNA expressions of calcium channels subunits Cav1.2,Calm,RyR2 and potassium channel Kir6.2 were analyzed by FQ-PCR method. RESULTS Laser confocal microscopy revealed that H2O2 obviously caused calcium overload in cardiomyocytes, giving rise to 49.37% fluorescence increase in intracellular calcium compared with the control group(P<0.01). However,natrin 5,25 and 125 mg·L-1 resulted in 27.52%, 12.71% and 5.15% fluorescence increase in intracellular calcium,respectively,compared with the control group(P<0.01). Moreover, the PCR results showed that the mRNA expressions of Cav1.2, Calm and RyR2 in the myocardial cells treated with H2O2 were increased 2.78,2.26,and 5.34 times as compared with the control group,while Kir6.2 displayed a 1.79-fold expression level(P<0.01). By contrast, the combination of natrin and verapamil significantly decreased the mRNA expression of Cav1.2,Calm and RyR2,compared to the H2O2-treated group(P<0.01). Meanwhile,the expression of Kir6.2 was considerably higher than that of the H2O2-treated group(P<0.05). CONCLUSION Natrin can reduce the intracellular calcium overload of cardiomyocytes induced by H2O2 and shows a protective effect against oxidative damage for cardiomyocytes. The possible mechanism is that natrin can decrease the mRNA expression of Cav1.2,Calm,RyR2 and increase the expression of Kir6.2 of the H2O2-induced cardiomyocytes.
ABSTRACT
To study the chemical constituents of Drynariae Rhizoma, nine phenolic acids were isolated from a 70% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, ODS, Sephadex LH-20, and semi-preparative HPLC. By spectroscopic techniques including 1H NMR, 13C NMR, 2D NMR, and HR-ESI-MS, these compounds were identified as 4, 4'-dihydroxy-3, 3'-imino-di-benzoic acid (1), protocatechuic acid (2), gallic acid (3), p-hydroxybenzoic acid (4), (E)-caffeic acid (5), ethyl trans-3, 4-dihydroxycinnamate (6), caffeic acid 4-O-beta-D-glucopyranoside (7), p-coumaric acid 4-O-beta-D-glucopyranoside (8), and 23(S)-12-O-caffeoyl-12-hydroxyllauric acid glycerol ester (9), separately. Among them, 1 and 9 are new compounds, and 3, 4, and 6 were isolated from Drynaria species for the first time.