UPLC-Q/TOF-MS based metabonomics revealed protective effect of Terminalia chebula extract on ischemic stroke rats

Terminalia chebula(TC),a kind of Combretaceae,is a widely used herb in India and East Asia to treat cerebrovascular diseases.However,the potential mechanism of the neuroprotective effects of TC at the metabonomics level is still not unclear.The present study focused on the effects of TC on metabonomics in stroke model.In our study,rats were divided randomly into Sham,Model,and TC groups.The TC group were intragastricly administered with TC for 7 d after middle cerebral artery occlusion(MCAO)operation.The sham and the model groups received vehicle for the same length of time.Subsequently,the neuroprotective effects of TC were examined by neurological defects evalua⁃tion,infarct volume assessment,and identification of biochemical indicators for antioxidant and anti-inflammatory activi⁃ties.Further,metabonomics technology was employed to evaluate the endogenous metabolites profiling systematically.Consist to results of biochemical and histopathological assays,pattern recognition analysis showed a clear separation of the Model and the Sham group,indicating a recovery impact of TC on the MCAO rats.Moreover,12 potential biomarkers were identified in MCAO Model group,involved in energy(lactic acid,succinic acid,and fumarate),amino acids(leucine,alanine,and phenylalanine)and glycerophospholipid[PC(16∶0/20∶4),PC(20:4/20:4),LysoPC(18:0)and LysoPC(16:0)]metabolism,and other types of metabolism(arachidonic acid and palmitoylcarnitine).Notably,we found that metabolite levels of TC group were partially reversed to normal.In conclusion,TC could ameliorate MCAO rats by intervening with energy metabolism(glycolysis and TCA cycle),amino acid metabolism,glycerophospholipid metabolism and other types of metabolism.

Termitomenins F and G,Two New Lignan Glucosides from Terminalia chebula var.tomentella(Kurz)C.B.Clarke

The extensive chemical investigation on the branches and leaves of Terminalia chebula var.tomentella(Combretaceae)led to the isolation of two new lignan glucosides with a furofuran skeleton,termitomenins F(1)and G(2).In addition,19 known compounds including five lignan glucosides(3-7),six hydrolyzable tannins(8-13)and eight simple phenolics(14-21)were also identified.Their structures were determined by comprehensive spectroscopic analyses.It is noted that 8 and 9 were C-glycosidic hydrolyzable tannins with one hexahydroxydiphenoyl and one gallagyl group linked to an open-chain glucosyl C-1/O-2/O-3 and O-4/O-6,respectively,which were rarely found in plants.Nine known compounds,6-9,13,and 18-21,were procured from the titled plant for the first time,while 3-5,10-12 and 14-17 were also found in the fruits.Notably,the known hydrolyzable tannins 8-13 exhibited strongerα-glucosidase inhibitory activities with IC_(50) values ranging from 0.10 to 3.12μM,than the positive control,quercetin(IC_(50)=9.38±0.33μM).

Antimicrobial potential of Terminalia chebula Retz. fruit extracts against ear pathogens

AIM： To evaluate the antimicrobial potential of Terminalia chebula （ T. chebula） extracts against pathogens causing otitis externa and compare it with ear drops.METHODS： Four different extracts, methanol, ethanol, acetone and aqueous （hot and cold） extracts, from the T. chebula were tested for their antimicrobial activity through the agar well diffusion method and minimum inhibitory concentration （MIC）/minimum bactericidal concentration （MBC） values were determined through the macrodilution broth method against six different microorganism, five bacterial （one gram positive and four gram negative） and one yeast.RESULTS： Organic and aqueous fruit extracts displayed activity against all fve tested bacterial ear pathogens with a maximum zone of inhibition of 31.6 mm against Staphylococcus aureus , followed by Acinetobacter sp. （24.6 mm）, Pseudomonas aeruginosa （23.6 mm）, Proteus mirabilis （21 mm） and Escherichia coli （19.3mm）. Of the four solvents evaluated, acetonic fruit extract of T. chebula was found to be best. The MIC values ranged between 0.78 mg/mL and 50 mg/mL for the different bacterial ear pathogens and MBC values ranged between 1.56 mg/mL and 50 mg/mL. The acetonic fruit extract showed larger inhibition zones compared to the herbal ear drops, Kan pip with lowest MIC of 0.78 mg/mL and MBC of 1.56 mg/mL.CONCLUSION： Acetonic extract of T. chebula fruit may be used to treat otitis externa. However, more detailed studies, such as in vivo testing and pharmacokinetics properties, are needed to determine its therapeutic potential.

In-vitro and In-vivo Anti-trypanosomal Activity of Terminalia chebula Retz Dried Fruits

African trypanosomosis had caused lots of havocs to both humans and animals over a century with successes and failure in curtailing it. This study was aimed at screening medicinal plant, Terminalia chebula dried fruits against Trypanosoma evansi for trypanocidal activity. Twenty grams of powdered Terminalia chebula dried fruits was cold extracted with methanol. Obtained MPE （methanolic plant extract） was in vitro tested against Trypanosoma brucei （1 × 10^6 trypanosomes/mL of the medium in each ELISA plate wells） at concentrations （250～1,000 μg/mL） on Vero cells grown in DMEM （Debecco＇s Modified Eagle Medium） in appropriate conditions for trypanocidal activity. In-vitro cytotoxicity test of MPE of Terminalia chebula was conducted on Vero cells grown in DMEM. In-vivo assay for trypanocidal activity, each mouse was inoculated with 1 × 10^4/mL of trypanosomes and treated （48 h post inoculation） with MPE of Terminalia chebula at concentrations （12.5, 25, 50, 100 and 200 mg/kg body weight） were administered at dose rate of 100 BL per mouse via intraperitoneal route to different groups of mice, 6 mice per concentration. In-vitro cytotoxicity test was done on Veto cells at concentrations （1.58～100 μg/mL） of MPE of Terminalia chebula. Results of in-vitro trypanocidal activity varied from immobilization, reduction and to the killing of the trypanosomes. At 250 μg/mL ofMPE ofTerminalia chebula dried fruits, there was significant trypanocidal activity at 4 h of incubation and trypanosomes were not detected in corresponding ELISA plate wells at 5 h of incubation, which was statistically equivalent to reference drug, diminazine aceturate （50 μL/mL） at 4 h of incubation. Results of in-vivo trypanocidal activity revealed that at concentrations （l 2.5～25 mg/kg body weight） of MPE of Terrninalia chebula, mice in these groups survived for 6 days. While at 50 and 100 to 200 mg/kg body weight, mice in these groups survived up to 7 and 8 days, respectively. In-vitro cytotoxicity test showed that all concentrations of MPE of Terminalia chenula and diminazine aceturate were cytotoxic to cells except at 1.56 μL/mL and 6.25 μL/mL. In conclusion, MPE of Terminalia chebula dried fruits possessed trypanocidal compounds. Further study （bioassay-guided purification） is required to know the full potential of Terrninalia chebula as future trypanocide candidate.

Effect of Terminalia chebula Retz. extraction with water on Staphylococcus epidermidis activity and its biofilm formation

Background:Based on modern pharmacological studies,Terminalia chebula Retz.exhibits antimicrobial activity against a variety of bacteria.Previously,we found Terminalia chebula Retz.exhibited excellent antibacterial activity against Malassezia restricta.Methods:We determined the minimal inhibitory concentration(MIC)and minimum bactericidal concentration of Terminalia chebula Retz.extraction with water(TRW)against Staphylococcus epidermidis(including Staphylococcus epidermidis 1-15)using the microdilution method.Staphylococcus epidermidis 1(SE11),which was the most sensitive to TRW,was selected as the test bacterium for subsequent experiments.The time-kill curve of TRW on SE11 was generated using the viable count method.Further,an in vitro biofilm model of SE11 was constructed using the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide test,semi-quantitative crystal violet experiment,and scanning electron microscopy.The effects of TRW on the total amount of biofilm formation,the number of viable biofilm bacteria,and biofilm microstructure of SE11 were also determined using a semi-quantitative crystal violet experiment,viable count method,and scanning electron microscopy.Finally,the electrical conductivity and soluble protein content of the SE11 suspensions were determined.Results:The average MIC of TRW against SE11 was 0.75±1.09 mg/mL.TRW(1/2MIC and 2MIC)and zinc pyrithione(1/2MIC and 2MIC)had no significant effect on total biofilm inhibition in the adhesion stage(P>0.05)and the aggregation stage(P>0.05).Further,TRW(1/2MIC and MIC)and zinc pyrithione(1/2MIC and MIC)had no significant effect on viable biofilm bacteria in the adhesion stage(P>0.05)and aggregation stage(P>0.05).TRW destroyed the integrity of the SE11 cell membrane,resulting in leakage of intracellular substances.Conclusion:TRW inhibits SE11 biofilm formation and is similar to zinc pyrithione in the adhesion and aggregation stages,which provides a theoretical basis for its application in the field of antimicrobial additives.

Terminalic Acid, a New Tannin from the Fruit of Terminalia chebula

Terminalic acid 1, a new hydrolysable tannin, was isolated from the fruit of Terminalia chebula. Its structure was found to be 1-O-galloyl-2,4-chebuloyl-beta-D-glucopyranose by chemical method and H-1-NMR, C-13-NMR, H-1-H-1 COSY, HMQC, HMBC and FAB-MS spectral analyses.

Extraction and Purification of Depigmenting Agents from Chinese Plants

Depigmenting agents were solvent-extracted and purified by preparative and analytical HPLC from three Chinese plants; Chrysanthemum morifolium Ramat（ Xizang Caijuhua）, Rhodiola sachalinensis, and Terminalia chebula Retzius. Four fractions obtained from the ethyl ether layer of C. m. Rama and two fractions from the ethyl acetate layer of Rhodiola salientness show depigmenting effects. At 8 200, the ethyl acetate layers of Chrysanthemum morifolium Ramat, Rhodiola sachalinensis and the methanol extract of Terminalia chebula Retzius, can inhibit the melanin production of mouse B16 melanoma cells by 92%, 60% and 90%, respectively, whereas 46% inhibition was observed by commercially available depigmenting agents（arbutin）. These results show the potential of these three Chinese plants as a novel resource for depigmenting agents in the cosmetic industry.

The changes of polyphenolic contents in two kinds of Terminalia chebula Retz.traditional Tibetian processing methods

Taking Terminalia chebula Retz. as the control product, the polyphenolic contents of the processed products in Tibetian, Rubia cordifolia L. processed Terminalia chebula Retz. and Euphorbia fischeriana Steud processed Terminalia chebula Retz. were analyzed by HPLC. Polyphenolic contents increased from 4.54% to 5.69% and 7.46%, respectively, which may lead to the change in their pharmacological effects.

Effects of different processing methods in Tibet on content of chebulinic acid in Terminalia chebula Retz.

To study the change of the content of chebulinic acid in Terminalia chebula Retz. by different processing methods in Tibet. We taking chebulinic acid as an indicator, the contents in Terminalia chebula Retz. and its processed products were analyzed by using HPLC. Also, the change rule was explored. Compared with Terminalia chebula Retz., the contents of chebulinic acid have decreased after processing, declining 50.18% for the Rubia cordifolia L. processed product, while 11.2% reduce for the Euphorbia fischeriana Steud processed product. After the specific preparation process in Tibet, the contents of chebulinic acid have reduced significantly, which may lead to different pharmacological effects.

Enhanced Antioxidant Capacities in vivo Caused by the Change in Key Constituent of Terminalia chebula Retz. Treated with Tibet Traditional Process

In order to investigate the chemical-biological relationship caused by the special processing technology,the constituent＇s content and antioxidant capacity in vivo of Euphorbia fischeriana Steud processing Terminalia chebula Retz. have been tested. Taking T. chebula Retz. as control,the content of gallic acid is analyzed with high performance liquid chromatography（HPLC） method. The results show that the increase of gallic acid concentration from 4.54% to 7.46% leads to enhancement of the antioxidant capacities in vivo of the E. fischeriana Steud processing T. chebula Retz. The significant antioxidant capacity in vivo（p〈0.05） of the processed product possesses is better than the unprocessed herb. Its superoxide dismutase（SOD）and malondia-ldehyde（MDA） contents in serum,the MDA content in liver can match the effects of vitamine E when administered at high dosage.

诃子活性成分诃子酸联合运动对T2DM大鼠糖脂代谢和铁死亡的影响

本研究旨在探讨诃子(Terminalia chebula)的活性成分诃子酸(chebulinic acid,CA)联合运动对2型糖尿病(T2DM)大鼠糖脂代谢和铁死亡的影响。通过高脂饮食联合链脲佐菌素(STZ)成功构建T2DM大鼠模型后,大鼠被随机分为七组。实验结果显示,与单独运动或仅给予高剂量诃子酸治疗的T2DM大鼠相比,诃子酸与运动联合治疗的大鼠其血糖、胰岛素、血脂等指标均得到明显改善,同时胰腺形态也有所恢复。此外,诃子酸和运动的结合还显著提高了抗氧化酶的活性,并减少了铁死亡相关的蛋白表达。本研究结果初步表明,诃子酸联合运动不仅可以有效改善T2DM大鼠的糖脂代谢,还可以抑制氧化应激和铁死亡,为防治T2DM提供了一个潜在的综合策略。

Scientific Validation of the Ethnomedicinal Properties of the Ayurvedic Drug Triphala:A Review

Triphala, a herbal formula composed of the three fruits of Terminalia chebula Retz. （Haritaki, Family： Combretaceae）, Terminalia bellirica Roxb. （Bibhitaki, Family： Combretaceae） and Phyllanthus emblica Linn. or Emblica officinalis Gaertn. （Amalaki or the Indian gooseberry, Family： Euphorbiaceae） is considered to be a universal panacea in the traditional Indian system of medicine the Ayurveda. It has been described in the Ayurveda text as a ＂Ftasayana＂ and to rejuvenat the debilitated organs. Ayurvedic physicians use Triphala for many ailments but most importantly to treat various gastrointestinal disorders. Scientific studies carried out in the past two decades have validated many of the ethnomedicinal claims and researches have shown Triphala to possess free radical scavenging, antioxidant, antiinflammatory, antipyretic, analgesic, antibacterial, antimutagenic, wound healing, anticariogenic, antistress, adaptogenic, hypoglycaemic, anticancer, chemoprotective, radioprotective and chemopreventive effects. Clinical studies have also shown that Triphala was found to have good laxative property, to improve appetite and reduce gastric hyperacidity. Studies have also shown that Triphala was effective in preventing dental caries and that this effect was equal to that of chlorhexidine. The current review addresses the validated pharmacological properties of Triphala and also emphasizes on aspects that need further investigation for its future clinic application.