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Objective: Metabonomics was used to analyze and explore the biomarkers and possible mechanisms of liver and kidney subacute toxicity induced by garidi-5 in rats. Methods: Taking garidi-5 as the target drug and SD rats as the research objects, each administration group except the normal group was intragastric administration of the corresponding drug solution for 28 d. The serum, liver and kidney samples of rats were detected by metabolomics and characterized by principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) to identify the sensitive markers and metabolic pathways of liver and kidney subacute toxicity. Results: Metabolomics analysis showed that compared with the normal group (Z), the 52, 64 and 54 different metabolites were identified in the serum, liver and kidney samples of garidi-5 high dose group (GG), which were mainly enriched in ABC transporters, arginine and proline metabolism, nicotinate and nicotinamide metabolism, central carbon metabolism in cancer pathways. Conclusion: The preliminarily suggested that garidi-5 can damage the liver and kidney by affecting the ABC transporters, arginine and proline metabolism, nicotinate and nicotinamide metabolism pathways, etc. Trimethylamine N-oxide, L-pyroglutamic acid, glycine-betaine, xanthine, glutathione, L-leucine, cytidine, L-arginine, spermidine, urea, 5-aminovaleric acid, creatine, L-glutamic acid, 1-methylnicotinamide and S-adenosyl-L-methionine can be used as potential biomarkers of liver and kidney toxicity sensitivity.
ABSTRACT
Introduction. Garidi-5, a traditional medicine composed of 5 herbs including Terminalia chebula Retz. Aconitum Kusnezoffii Reichb., Acorus calamus L., Saussurea lappa L., and musk of Moschus moschiferus, has been used in traditional Mongolian medicine as an analgesic and antibacterial medicine. The present work was undertaken to evaluate the traditional drug Garidi-5 for its analgesic and anti-inflammatory activity.Materials and Methods. The method of Winter et al. was used to study acute inflammation. Rats in groups of five each were treated with vehicle, Garidi-5 (20, 80 and 200 mg/kg, p.o.) and Indometacin (10 mg/kg) one hour prior to Carrageenan injection. 0.1 ml of 1% Carrageenan was injected into the subplantar tissue of left hind paw of each rat. Swelling of carrageenan injected foot was measured at 0, 0.5, 2, 4 h using Plethysmometer (UGO Basile, Italy)). The right hind pawwas injected with 0.1 ml of vehicle.ResultsThe Garidi-5 (20, 80 and 200 mg/kg) significantly (P<0.01) inhibited carrageenan induced rat paw edema as compared to control group. Maximum inhibition of paw edema was observed with Garidi-5 (80 and 200 mg/kg) at 4 h when compared to the control group (Tab.1). In assay data, the TNF-α, IL-1β and IL-6 secretion in serum were highly elevated by carrageenan induction but administration of Garidi-5 signifi cantly reduced serum secretion of inflammatory mediators as compared to vehicle group (Tab. 2). ConclusionIn conclusion, Traditional drug Garidi-5 have anti-in flammatory properties. The potential efficacy of Garidi-5 to treat inflammation is based in a part on the hy pothesis that it will suppress the proinflammatory cytok ines resulting in less oedema.
ABSTRACT
Introduction. Garidi-5, a traditional medicine composed of 5 herbs including Terminalia chebula Retz., Aconitum Kusnezoffii Reichb., Acorus calamus L., Saussurea lappa L., and musk of Moschus moschiferus, has been used in traditional Mongolian medicine as an analgesic and antibacterial medicine. The present work was undertaken to evaluate the traditional drug Garidi-5 for its analgesic and anti-inflammatory activity.Materials and Methods. The method of Winter et al. was used to study acute inflammation. Rats in groups of five each were treated with vehicle, Garidi-5 (20, 80 and 200 mg/kg, p.o.) and Indometacin (10 mg/kg) one hour prior to Carrageenan injection. 0.1 ml of 1% Carrageenan was injected into the subplantar tissue of left hind paw of each rat. Swelling of carrageenan injected foot was measured at 0, 0.5, 2, 4 h using Plethysmometer (UGO Basile, Italy)). The right hind paw was injected with 0.1 ml of vehicle.Results. The Garidi-5 (20, 80 and 200 mg/kg) significantly (P<0.01) inhibited carrageenan induced rat paw edema as compared to control group. Maximum inhibition of paw edema was observed with Garidi- 5 (80 and 200 mg/kg) at 4 h when compared to the control group (Tab.1). In assay data, the TNF-α, IL-1β and IL-6 secretion in serum were highly elevated by carrageenan induction but administration of Garidi-5 signifi cantly reduced serum secretion of inflammatory mediatorsas compared to vehicle group (Tab. 2).Conclusion. In conclusion, Traditional drug Garidi-5 have anti-in flammatory properties. The potential efficacy of Garidi-5 to treat inflammation is based in a part on the hy pothesis that it will suppress the proinflammatory cytok ines resulting in less oedema.
ABSTRACT
BackgroundThe preparations of multi-component have been the subject of chemical study for a long time. Therefore, when compounding the preparations of multi-component in traditional medicine, their taste is cautiously relied on, as the power of the one medicine should not be subdued with the power of another. Additionally the properties of the components and their regulating effects on the body systems are also considered. Our research group has been carrying out tests for raw materials,which are contained in multi-component preparations. However, it is a necessity to conduct phytochemical study on multi-component preparations in order to isolate pure biological active compounds and to identify their structure as well as to quantify its amount by modern techniques of analysis.GoalThe aim of the present study was to isolate pure biological active substances from Mongolian traditional medicine Garidi-5 and to elucidate their structures, which was used in Mongolian traditional medicine for the treatment of inflammation and as a pain relieving remedy.Objectives:1. To isolate pure substances from Garidi-5 and carry out tests to identify and determine their structure2. To quantify the amount of biological active substances.Materials and MethodsMongolian traditional medicine Garidi-5 has been selected as a biological natural product for the study. Garidi-5 is a traditional Mongolian medicine consisting of 5 medicinal herbs, namely Terminalia chebula Retz., Aconitum Kusnezoffii Reichb., Acorus calamus L., Saussurea lappa L., and musk of Moschus moschiferus and manufactured in the Drug factory of Traditional Medical Science Technology and Production Corporation of Mongolia. In this research, in order to determine the total content of phenolic compound was used the Folin–Ciocalteu method, which based on performing dark blue color complex compound. Isolated substance identification was determined by the TLC, UV and IR spectrophotometric methods. Inaddition it was checked melting point of the isolated substance. Determination of Gallic acid30g Garidi-5 was macerated in 60ml 80% methanol at room temperature for 24 h. After extraction, the extract was concentrated and vacuum evaporated. Different solvents from hexane, chloroform, ethyl acetate and n-butanol were used for theexperiment. All the extracts collected, evaporated and chromatographed on Silica gel column. Future purification of active fractions on Silica gel with methanol yielded the compound G1 which was further characterized as Gallic acid. Total phenolic content was determined spectrophotometrically according to the Folin–Ciocalteu’s method with slight modification. Gallic acid was used as a standard phenolic compound. Briefly, 1 ml of extract solution contains 1 mg extracts, in a volumetric flask diluted with distilled water (46 ml). One ml of Folin-Ciocalteu reagent was added and the content of the flask mixed thoroughly. After 3 min, 3 ml of Na2CO3 (2%) was added and then the mixture was allowed to stand for 2 h with intermittent shaking. The absorbance was measured at 760 nm in a spectrophotometer. All measurements were performed in triplicate.ResultsIn this research, TLC method on silica gel plates was used in order to identify the biological active pure substances from Garidi-5. Preliminary TLC experiments indicated the presence of Gallic acid in Garidi-5, which was isolated by column chromatography by comparing with reference standard substance (Gallic acid). Gallic acid was determined in the solvent system benzole-ethyl acetateformic acid- acetone (5:5:2:0.5) in isolated substance (G1). It showed blue color, Rf =0.65, on TLC plate. [1] For the characterization of two samples it was carried out IR analysis for each. In the IR spectra of G1 and standard substance can be recognized by the following absorption frequency regions: 700-900 cm-1 for Car-H; 1000-1300 cm-1 for vibration of bonds in various oxygen containing groups, 1350-1470 cm-1 for vibrations of –CH, -CH2 and –CH3 groups; 1500-1630 cm-1 for skeletal vibrations of aromatic rings, >C=O bonds; 2800-2950 cm-1 for stretching vibrations of –CH, -CH2 and -CH3 groups in saturated aliphatic structures; and 3030-3350 cm-1 for stretching associated vibrations of -OH groups in aromatic rings and aliphatic structures. As a result it was revealed that both IR spectra of G1 and standard substances were similar. [3]Further for the characterization of two samples it was carried out UV analysis of each. In the UV spectra of G1 and standard substance can be recognized by the following absorption frequency regions: 260-280nm for benzole groups; 200-225nm for carbonic acids; 400-770nm >C=O bonds, which reveal the presence of Gallic acid. In addition, melting point of isolated substance G1 was analyzed and detected at 2410C, which was similar to the standard substance’s melting point. [4]Moreover, Mongolian traditional medicine Garidi-5 contains 24% of the biological active substance (total phenolic compounds). [2]Conclusions:As a result of current study on Mongolian medicine Garidi-5, it was isolated one essential substance from ethyl acetate fraction. The phytochemical analysis reveals the presence of Gallic acid in Garidi- 5, which was determined by thin layer chromatography, UV and IR spectrophotometric methods. Mongolian traditional medicine Garidi-5 contains 24% of the biological active substance. Thus, the isolation of Gallic acid from multi-component preparations and identification of its structure was first phytochemical study conducted in our laboratory.