RP-HPLC测定桑椹中芦丁的含量

目的:优化桑椹中芦丁的提取条件并建立芦丁的HPLC测定方法。方法:采用单因素考察法优化桑椹中芦丁的提取条件,采用HPLC法测定芦丁的含量,色谱柱为Agilent Zobax C18(4.6 mm×250 mm,5μm),以乙腈-0.1%磷酸(19∶81)为流动相,流速1.0 mL.min-1,检测波长360 nm,柱温30℃。结果:芦丁最佳提取条件为料液比1∶25,50%乙醇回流60 min,芦丁在0.1～2.0μg有良好线性关系(r=0.999 9)。平均回收率为102.4%,RSD 1.8%(n=6)。结论:该方法简便、准确、可靠,可作为桑椹中芦丁含量测定的方法。     

Vascular effects of Tanacetum vulgare L. leaf extract: in vitro pharmacological study

Aim of the study

Tanacetum vulgare L. (Asteraceae/Compositae) is principally used in traditional Moroccan medicine as antihypertensive remedy. The aim of the present study was to investigate the in vitro vascular activity of the aqueous extract of Tanacetum vulgare L.

Materials and Methods

The activity of Tanacetum vulgare L. extract was tested on contractile response of Wistar rat aorta to high KCl and noradrenaline and on endothelium-dependent relaxation evoked by acetylcholine.

Results

The addition of Tanacetum extract during the plateau phase of noradrenaline-evoked contraction produced a rapid relaxation that reached a maximum of 30% of the contraction and was suppressed by the NO synthase inhibitor N^G-nitro-l-arginine. At higher extract concentrations this rapid relaxation was followed by a slowly developing, N^G-nitro-l-arginine-resistant, relaxing effect. Tanacetum extract also depressed KCl-evoked contraction by 30% at maximum. This effect was abolished in the presence of N^G-nitro-l-arginine. The endothelium-dependent relaxation induced by acetylcholine was depressed in the presence of Tanacetum extract in the bathing solution.

Conclusion

:This study indicates that the aqueous extract of Tanacetum possesses NO-mediated and NO-independent vasorelaxing properties in vitro.     

Evaluation of selected tropical seaweeds for in vitro anti-trichomonal activity

Ethnopharmacological relevance

Human parasitic infections are a serious problem in tropical and sub-tropical developing countries. Trichomoniasis, responsible for the annual infection of 180 million people, is a common sexually transmitted disease caused by the protozoan Trichomonas vaginalis. Traditionally seaweeds have been used in folk medicine by coastal people in Asia and the Caribbean to treat parasitic infections and are a valuable source of novel anti-trichomonals.

Aim of the study

In our search for therapeutical alternatives to anti-protozoal chemotherapy, we collected a selection of 25 tropical seaweeds (12 Rhodophyta, 5 Phaeophyta and 8 Chlorophyta) from the coast of Yucatan (Mexico) in order to undertake ethnopharmacological and chemotaxonomic investigations.

Materials and methods

Organic algal extracts were tested for their anti-trichomonal properties on the growth inhibition of Trichomonas vaginalis. The cytotoxicity of seaweed extracts on mammal cell lines was also assessed.

Results

The results indicated that 44% of the seaweeds studied had high to moderate anti-trichomonal activity. Lobophora variegata and Udotea conglutinata showed the maximal anti-trichomonal activity with IC₅₀ values of 1.39 and 1.66 μg/ml, respectively, with good selectivity.

Conclusions

Lobophora variegata and Udotea conglutinata demonstrated promising anti-trichomonal potential and have been selected for further bio-guided fractionation and isolation of active anti-trichomonal compounds.     

Anticancer potential of aqueous extract of alocasia macrorrhiza against hepatic cancer in vitro and in vivo

Ethnopharmacological relevance

Alocasia macrorrhiza has been used as a folk medicine for cancer treatment in the Southwest of China.

Aim of the study

The purpose of this study is to confirm the anticancer activity of aqueous extract of alocasia macrorrhiza against hepatic cancer and to elucidate its mechanism of action.

Materials and methods

Human normal liver cells and hepatocellular carcinoma cells were tested in vitro for cytotoxicity, colony formation inhibition, EdU incorporation, AO/EB staining apoptotic cells, apoptotic DNA fragmentation, and cell cycle distribution in response to alocasia macrorrhiza extract. The mRNA and protein expressions of PPARγ, Cyclin D1, Rb, P21, Bax, Bcl-2 and caspase-3 were detected through RT-PCR and Western blotting; the tumor growth inhibition in vivo was tested by oral administration of the extract.

Results

Alocasia macrorrhiza aqueous extract exhibited proliferation inhibition and apoptosis effects on human hepatocellular carcinoma cells in vitro, inhibited hepatoma growth in vivo.

Conclusion

Alocasia macrorrhiza extract has potential cytotoxic and apoptotic effect on human hepatocellular carcinoma cells and inhibits hepatoma growth in vivo, its mechanism of action might be associated with the inhibition of DNA synthesis, cell cycle (G₀/G₁) arrest, apoptosis induction through up-regulation the mRNA and protein expressions of PPARγ, Rb, Bax and capase-3genes and down-regulation of the expressions of Cyclin D1 and Bcl-2 genes.     

In vitro and in vivo antioxidant effects of the ethanolic extract of Swertia chirayita

Ethnopharmacological relevance

Swertia chirayita, a medicinal herb endemic to the Tibetan region, is used as a special remedy for liver disorders. The hepatoprotective activity of its plant extracts has been associated with its antioxidant activity. This paper aims to investigate the in vitro and in vivo antioxidant effects of Swertia chirayita extracts (SCE).

Materials and methods

Antioxidant ability of Swertia chirayita was investigated by employing several established in vitro methods. In vivo antioxidant activity was tested against CCl₄-induced toxicity in mice. The levels and activities of malondialdehyde (MDA) and antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH), were then assayed using standard procedures.

Results

SCE exhibited strong antioxidant ability in vitro. The liver and kidney of CCl₄-intoxicated animals exhibited a significant (p < 0.001) decrease in SOD, CAT, and GSH levels. Additionally, these organs exhibited a significant (p < 0.001) increase in MDA level. CCl₄ did not exhibit toxicity on mice treated with SCE and Vitamin E. The effects of Swertia chirayita (three dosages) were comparable to those of Vitamin E, except in MDA level in the liver and GSH level in the kidney (p < 0.05).

Conclusion

This study suggests that the ethanolic extract of Swertia chirayita possesses in vitro and in vivo antioxidant effects. This supports the traditional use of Swertia chirayita in Tibetan medicine to cure liver diseases.     

In vitro and in vivo antioxidant activity of the ethanolic extract from Meconopsis quintuplinervia

Aims of the study

Meconopsis quintuplinervia, a medicinal herb endemic to the Tibetan region, is used to treat hepatitis. The aim of this study is to evaluate the antioxidant potential of the ethanolic extract of this herb using different assays.

Materials and methods

The antioxidant capacity of Meconopsis quintuplinervia was investigated using various established in vitro systems. An in vivo study of carbon tetrachloride (CCl₄)-induced antioxidant activity in mice was also conducted by examining the levels of malondialdehyde (MDA) and the activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH).

Results

The extract showed strong in vitro antioxidant ability. In the in vivo study, CCl₄-induced oxidative stress caused significant decreases in the SOD, CAT, and GSH levels and a significant increase in the MDA level, most of which were significantly reversed (except for SOD in the liver.) by treatment with the extract and standard Vitamin E.

Conclusion

This study clearly indicates that the ethanolic extract of Meconopsis quintuplinervia is a valuable source of natural antioxidants. These findings provide scientific support for the traditional use of this herb as a Tibetan medicine for liver diseases.     

Evaluation of anti-inflammatory potential of Chloranthus erectus (Buch.-Ham.) Verd. leaf extract in rats

Aim of the study

Chloranthus erectus (Buch.-Ham.) Verdcourt (Chloranthaceae) is a shrub native to tropical and temperate zone of Eastern Himalaya of India and South-East Asia and have traditionally been used as a folklore medicine to treat localised swelling, joint pain, skin inflammation, fever and bodyache. In this study, an attempt has been made to demonstrate the anti-inflammatory activity of methanol extract obtained from Chloranthus erectus leaves (MECEL) in acute, sub-acute and chronic mouse models.

Materials and methods

Inflammation in the hind paw of Wistar albino rat was induced by carrageenan, histamine and serotonin, and tissue granuloma pouch was induced by cotton pellet method. Anti-inflammatory drug—phenylbutazone was used as standard drug for comparison.

Results

In acute carrageenan-induced rat hind paw edema, oral administration of MECEL at 200 mg/kg produced significant inhibition of edema by 38.34% (p < 0.01) while the histamine- and serotonin-induced sub-acute model, the inhibition of paw edema reached 52.54% (p < 0.001) and 25.5% (p < 0.01), respectively. In a 7-day study, MECEL at 20 and 50 mg/kg produced significant suppression of cotton pellet-induced tissue granuloma formation in rats.

Conclusions

This preliminary study revealed that the methanol extract of Chloranthus erectus exhibited significant anti-inflammatory activity in the tested models, and may provide the scientific rationale for its popular folk medicine as anti-inflammatory agent.     

Genotoxic assessment of Rubus imperialis (Rosaceae) extract in vivo and its potential chemoprevention against cyclophosphamide-induced DNA damage

Ethnopharmacological relevance

Rubus imperialis Cham. Schl. (Rosaceae) is frequently used in traditional medicine as hypoglycemic, antinociceptive and antiviral remedy.

Materials and methods

Swiss albino mice were distributed in eight groups for acute treatment with Rubus imperialis extract (24 h). The extract doses selected were 50, 250 and 500 mg/kg b.w. administered by gavage alone or plus to CPA (50 mg/kg b.w.) administered by intraperitoneal injection. Control groups were treated in a similar way. Analyses were performed using the comet assay, on leukocytes (collected 4 and 24 h after treatment) and liver (collected 24 h after treatment), and using the micronucleus test (MN) in bone marrow cells. Cytotoxicity was assessed by scoring 200 consecutive polychromatic (PCE) and normochromatic (NCE) erythrocytes (PCE/NCE ratio).

Results and conclusion

The main compounds identified in the Rubus imperialis extract were saponins and steroidal compounds, with niga-ichigoside and tormentic acid being the major compounds. Tested doses of Rubus imperialis extract showed no genotoxic effects on leukocytes from peripheral blood or liver cells by the comet assay. However, the MN test showed an increase in the frequency of micronucleated cells at the two higher doses tested, indicating that this extract has clastogenic/aneugenic effects on bone marrow cells at higher doses. On the other hand, for all cells evaluated, the three tested doses of the Rubus imperialis extract promoted inhibition of DNA damage induced by CPA. Despite the chemoprevention observed, the clastogenicity/aneugenicity observed suggested caution about either continuous or high-dose usage of Rubus imperialis aerial parts extract by humans.     

Typhonium flagelliforme inhibits cancer cell growth in vitro and induces apoptosis: an evaluation by the bioactivity guided approach

AIM OF THE STUDY: Typhonium flagelliforme (Lodd.) Blume (Araceae) is a Malaysian plant used locally to combat cancer. In order to evaluate its antiproliferative activity in vitro and to possibly identify the active chemical constituents, a bioactivity guided study was conducted on the extracts of this plant. MATERIALS AND METHODS: The active extracts of Typhonium flagelliforme were fractionated by flash column chromatography and each fraction was evaluated for antiproliferative activity using MTT assay. The apoptotic effect of the active fraction was determined microscopically and by using TUNEL colorimetric assay. GC-MS and NMR were used to determine the chemical constituents of this active fraction. RESULTS: Several fractions of the hexane and dichloromethane extracts were found to inhibit the growth of NCI-H23 non-small cell lung carcinoma cell line significantly, with IC(50)<15 microg/ml. However, most of these active fractions were also found to inhibit the growth of non-tumorigenic BALB/c 3T3 mouse fibroblast cell line except for fraction 21 of the dichloromethane extract (D/F21). This particular fraction was not only less cytotoxic to the non-tumorigenic cells, where the IC(50) was 48.6 microg/ml compared to IC(50) 7.5 microg/ml for NCI-H23, but it was also found to induce apoptosis in the cancer cell line. GC-MS analysis revealed that D/F21 contains hexadecanoic acid, 1-hexadecene, phytol and a derivative of phytol. The presence of non-saturated fatty acids in this fraction was confirmed by nuclear magnetic resonance spectroscopy. CONCLUSIONS: D/F21 was found to be the active and cancer cell line specific fraction of Typhonium flagelliforme. Its major chemical constituents had been determined spectroscopically.     

Hypotensive effect of aqueous extract of Averrhoa carambola L. (Oxalidaceae) in rats: an in vivo and in vitro approach

Aim of the study

Averrhoa carambola L. (Oxalidaceae) leaves are used in Brazilian traditional medicine to treat hypertension. This study was conducted to evaluate the hypotensive effect of the aqueous extract of Averrhoa carambola (AEAc) and its underlying mechanisms in the isolated rat aorta.

Materials and methods

The effect of AEAc on the mean arterial pressure (MAP) was determined in vivo in anesthetized rats. In vitro, thoracic aortic rings were isolated and suspended in organ baths, and the effects of AEAc were studied by means of isometric tension recording experiments. In HPLC analysis, the fingerprint chromatogram of AEAc was established.

Results

In normotensive rats, AEAc (12.5-50.0 mg/kg, i.v.) induced dose-dependent hypotension. In vitro, AEAc caused a depression in the E_max response to phenylephrine without a change in sensibility. Also, in a depolarized Ca²⁺-free medium, AEAc inhibited CaCl₂-induced contractions and caused a concentration-dependent rightward shift of the response curves, indicating that AEAc inhibited the contractile mechanisms involving extracellular Ca²⁺ influx.

Conclusions

These results demonstrate the hypotensive effects of AEAc, and these effects may, in part, be due to the inhibition of Ca²⁺, which supports previous claims of its traditional use.     

Antidiabetic potential of unripe Carissa carandas Linn. fruit extract

Ethnopharmacological relevance

Carissa carandas commonly known as Karanda have a long history of use in traditional system of medicine. It is used by tribal healers of Western Ghat region of Karnataka as hepatoprotective and antihyperglycemic. However, no scientific data is available to validate the folklore claim. The present study has been designed to evaluate its unripe fruit for the antidiabetic activity.

Aim

In the present study, methanol extract of unripe fruits and its fractions were studied for its antidiabetic potential.

Materials and methods

The methanol extract and its fractions were screened for antidiabetic activity in alloxan induced diabetic rats. The polyphenolic, flavonoid and flavanone contents of methanolic extract and its fractions were also determined and correlated with its antidiabetic activity.

Results

The experimental data indicated that the methanol extract and its ethyl acetate soluble fraction has significantly lowered the elevated blood glucose levels by 48% (p < 0.001) and 64.5% (p < 0.001) respectively at dose level of 400 mg/kg per oral after 24 h as compared to diabetic control. In order to assess the role of polyphenolic components in the relevant activity, polyphenolic and flavonoid contents were determined. The polyphenolic and flavonoid content of methanol extract and its ethyl acetate soluble fraction were found to be 15.8 ± 1.2 mg and 18.55 ± 0.34 mg (gallic acid equivalent/g extract) and flavonoid content 2.92 ± 0.03 mg and 1.534 ± 0.30 mg (rutin equivalent/g extract) respectively.

Conclusion

The increased antidiabetic potential of ethyl acetate fraction over methanol extract is due to its partial purification achieved by fractionation which resulted in increase in degree of polymerization and segregation of secondary metabolites.     

Ficus spp. (fig): ethnobotany and potential as anticancer and anti-inflammatory agents

This review explores medieval, ancient and modern sources for ethnopharmacological uses of Ficus (fig) species, specifically for employment against malignant disease and inflammation. The close connection between inflammatory/infectious and cancerous diseases is apparent both from the medieval/ancient merging of these concepts and the modern pharmacological recognition of the initiating and promoting importance of inflammation for cancer growth. Also considered are chemical groups and compounds underlying the anticancer and anti-inflammatory actions, the relationship of fig wasps and fig botany, extraction and storage of fig latex, and traditional methods of preparing fig medicaments including fig lye, fig wine and medicinal poultices.     

Evaluation of the genotoxicity of Orthosiphon stamineus aqueous extract

Aim of the study

Orthosiphon stamineus, Benth, also known as Misai Kucing in Malaysia and Java tea in Indonesia, is traditionally used in Southeastern Asia to treat kidney dysfunctions, diabetes, gout and several other illnesses. Recent studies of Orthosiphon stamineus pharmacological profile have revealed antioxidant properties and other potentially useful biological activities thereby lending some scientific support to its use in folk medicine. So far the genotoxicity of Orthosiphon stamineus extracts has not been evaluated. In this study the genotoxic potential of Orthosiphon stamineus aqueous extract was investigated by the Salmonella/microsome mutation assay and the mouse bone marrow micronucleus test.

Materials and methods

Chemical composition of Orthosiphon stamineus aqueous extract was analyzed by High Performance Liquid Chromatography-Diode Array Detector (HPLC-DAD). The Salmonella/microsome assay (TA97a, TA98, TA100 and TA1535; plate incorporation method) was performed in the presence or in the absence of extrinsic metabolic activation (S9 mixture). In the mouse micronucleus assay, Orthosiphon stamineus extract was administered by gavage (0, 500, 2000 and 4000 mg/kg body weight/day for 3 days) to male and female Swiss Webster mice (N = 6 per dose per sex) and bone marrow cells were harvested 24 h after the last dose. Ethoxy-resorufin-O-dealkylase (EROD) and benzyloxy-resorufin-O-dealkylase (BROD) activities were determined in mouse liver microsomes.

Results

The chemical analysis revealed that the Orthosiphon stamineus extract contained flavonoids (sinensetin, eupatorin), caffeic acid, and rosmarinic acid (44.00 ± 1.879 μg/mg), the latter seemed to be one of its major constituent. Tested at doses up to 5000 μg/plate, the Orthosiphon stamineus extract was not toxic to Salmonella tester strains and did not increase the number of revertant colonies over the background incidence. In the mouse bone marrow assay, the extract did not alter the polychromatic:normochromatic erythrocytes (PCE:NCE) ratio, nor did it increase the incidence of micronucleated polychromatic erythrocytes (MNPEs). No overt toxicity and no change of CYP1A (EROD) and 2B9/10 (BROD) activities were noted.

Conclusions

Based on the aforementioned findings, it is concluded that the use of Orthosiphon stamineus in the traditional medicine poses no genotoxic risk.     

Evaluation of in vivo wound healing activity of methanol extract of Achyranthes aspera L

Ethnopharmacological relevance

The leaves of Achyranthes aspera L. (Amarenthacea) has been used traditionally for the treatment of wound in various parts of Ethiopia. However, the plant has not been explored scientifically for its wound healing activity. Therefore, this study was designed to investigate the wound healing activity of methanol extract of Achyranthes aspera L. leaves in rats.

Materials and methods

Incision and excision wounds were inflicted on albino rats of either sex, under diethyl ether anesthesia. Group I served as positive control and was treated with 1% silver sulphadiazine, group II, III, IV treated with simple ointment containing 2.5%, 5% and 10% (w/w) methanol extract of the leaves of Achyranthes aspera L., respectively, whereas group V served as negative control and was treated with simple ointment. All the animals were treated topically once a day. Wound healing potential was assessed with excision and incision wound model. Excision wound model was used to assess the change in percentage contraction of wound, epithelization time, DNA content and histological features whereas rats inflicted with the incision wounds were used to determine breaking strength.

Results

Based on the results of percentage wound contraction, the DNA content and epithelization time, all groups of rats treated with methanol extract of the leaves of Achyranthes aspera L. showed significant (p<0.05) wound healing activity compared to group of rats treated with simple ointment (negative control) group. The difference in breaking strength was, however, significant (p<0.05) only for the 5% and 10% methanol extract of Achyranthes aspera (w/w) ointment treated groups. Histological evaluation showed well organized epidermal layer, increased number of fibrocytes, remarkable degree of neovascularization and epithelization which was comparable to the standard on the 21st day after treatment; especially in the 5% and 10% (w/w) extract treated group.

Conclusion

The present study provides a scientific rationale for the traditional use of the leaf extracts of Achyranthes aspera L. in the treatment of wound.     

Evaluation of anticancer activity of ethanol extract of Sesbania grandiflora (Agati Sesban) against Ehrlich ascites carcinoma in Swiss albino mice

Ethno pharmacological relevance

The plant Sesbania grandiflora (Fabaceae) is commonly known as “Sesbania” and “agathi” in ayurvedic system of medicine and reputed in the indigenous medicine in India. It is also known as “Agati Sesban” or “humming bird tree” in English. All parts of this unique plant are useful and have a wide spectrum of medicinal properties. The plant has various uses in folk and traditional medicines for headache, swellings, anemia, bronchitis, pains, liver disorders and tumors.

Aim of the study

The objective of the present study was to explore the anticancer activity of the ethanol extract of Sesbania grandiflora against Ehrlich ascites carcinoma (EAC)-bearing Swiss albino mice.

Materials and methods

Anticancer activity of ethanol extract of Sesbania grandiflora (EESG) of both leaves and flowers were evaluated in Swiss albino mice against Ehrlich Ascites Carcinoma (EAC) cell line at the doses of 100 and 200 mg/kg body weight intraperitoneally. The extracts were administered for 14 consecutive days. Twenty-four hours of last dose and 18 h of fasting, the mice were sacrificed and the anticancer effect of EESG was assessed by evaluating tumor volume, viable and nonviable tumor cell count, tumor weight, hematological parameters and biochemical parameters of EAC bearing host.

Results

Sesbania grandiflora extracts showed significant decrease in (p < 0.01) tumor volume, viable cell count, tumor weight and elevated the life span of EAC bearing mice. Hematological profile such as RBC, hemoglobin and lymphocyte count reverted to normal level in EESG treated mice. The extracts significantly (p < 0.05) decreased the levels of lipid peroxidation and significantly (p < 0.05) increased the levels of GSH, SOD and CAT.

Conclusion

The results showed that the ethanol extract of Sesbania grandiflora was effective in inhibiting the tumor growth in ascitic models and that is comparable to 5-Fluorouracil.     

Intracellular replication inhibitory effects of Galla Rhois ethanol extract for Brucella abortus infection

Ethnopharmacological relevance

Galla Rhois (GR) has long been applied in traditional Korean and Oriental medicine. Although GR has an anti-bacterial effect, the anti-bacterial mechanism and therapeutic efficiency of GR for intracellular parasitic Brucella infection are still unclear.

Aim of the study

The objective of this study was to investigate the antibacterial and therapeutic effects of GR ethanol extract (GRE), which is a natural antibacterial component for the treatment of Brucella abortus infection.

Materials and methods

The antibacterial activity of GRE towards Brucella abortus was evaluated by incubating Brucella abortus with GRE. Following treatment with GRE, Brucella abortus adherence, uptake, intracellular growth, and intracellular trafficking in macrophages were monitored. Mice were infected intraperitoneally with Brucella abortus and treated orally with GRE for 14 days, and then the weight and CFUs from each spleen were monitored.

Results

The viability of Brucella abortus was markedly decreased in a dose-dependent manner. Moreover, Brucella abortus internalization and intracellular growth within macrophages were reduced in GRE-treated cells. The number of bacteria that adhered to GRE-pretreated cells was significantly lower than that of untreated cells. With regards to intracellular trafficking, treatment with GRE augmented the colocalization of Brucella abortus-containing phagosomes with LAMP-1. GRE-treated mice showed considerably decreased weight and bacterial burdens in the spleen compared to untreated mice.

Conclusion

GRE exhibits antibacterial and protective effects on Brucella abortus in vitro and in vivo. These results highlight the beneficial effects of GRE in the prevention and treatment of brucellosis.     

In vitro antitrypanosomal and antileishmanial activity of plants used in Benin in traditional medicine and bio-guided fractionation of the most active extract

Ethnopharmacological relevance

The aim of the study was to evaluate the in vitro antitrypanosomal and antileishmanial activity of crude extracts of 10 plant species traditionally used in Benin to treat parasitic infections.

Materials and methods

For each species, dichloromethane, methanol and aqueous extracts were tested. Their antitrypanosomal and antileishmanial activities were evaluated in vitro on Trypanosoma brucei brucei (strain 427) (Tbb) and on promastigotes of Leishmania mexicana mexicana (MHOM/BZ/84/BEL46) (Lmm).

Results

The best growth inhibition was observed with the dichloromethane extracts of aerial parts of Acanthospermum hispidum DC. (Asteraceae) (IC₅₀ = 14.5 μg/ml on Tbb and 11.1 μg/ml on Lmm), twigs of Keetia leucantha (K. Krause) Bridson (syn. Plectronia leucantha Krause) (IC₅₀ = 5.8 μg/ml on Tbb), aerial parts of Byrsocarpus coccineus Schumach. & Thonn (syn. Rourea coccinea (Schumach. & Thonn.) Hook.f.) (IC₅₀ = 14.7 μg/ml on Tbb) and aerial parts of Carpolobia lutea G.Don. (IC₅₀ = 18.3 μg/ml on Tbb). All these extracts had a low cytotoxicity. It is not the case for the methanolic and water extracts of roots of Anchomanes difformis (Blume) Engl. (IC₅₀ = 14.7 and 13.8 μg/ml on Tbb) which were toxic at the same concentration range on WI38, human cells. A bio-guided fractionation of the most active extract of Keetia leucantha allowed to identify oleanolic acid and ursolic acid as responsible for the observed activities.

Conclusion

Our study gives some justification for antiparasitic activity of some investigated plants.     

Comparative “in vitro” evaluation of the antiresorptive activity residing in four Ayurvedic medicinal plants. Hemidesmus indicus emerges for its potential in the treatment of bone loss diseases

Ethnopharmacological relevance

Four Indian plants, traditionally used in Ayurvedic medicine: Asparagus racemosus Willd., Emblica officinalis Gaertn., Hemidesmus indicus R. Br., and Rubia cordifolia L. were selected on the basis of their ethnobotanical use and of scientific evidence that suggests a potential efficacy in the treatment of bone-loss diseases. The antiresorptive properties of the four plants have been investigated. The aim was to provide adequate evidence for the exploitation of natural compounds as alternative therapeutics for the treatment of diseases caused by increased osteoclast activity.

Materials and methods

Decoctions were prepared from dried plant material according to the traditional procedure and standardization by HPLC was performed using marker compounds for each species. Total polyphenols, flavonoids and radical scavenging activity of the decoctions were also determined. The bioactivity of the plant decoctions was evaluated in subsequent phases. (1) A cytotoxicity screening was performed on the mouse monocytic RAW 264.7 cell line to define the concentrations that could be utilized in the following step. (2) The antiresorptive properties of plant decoctions were compared with that of a “gold standard” drug (alendronate) by measuring osteoclastogenesis inhibition and osteoclast apoptosis. (3) The toxic effect on bone forming cells was excluded by evaluating the impact on the proliferation of osteogenic precursors (mesenchymal stem cells, MSC).

Results

All the decoctions inhibited osteoclastogenesis similarly to alendronate at the highest doses, but Hemidesmus indicus and Rubia cordifolia were also effective at lower concentrations. Apoptosis increased significantly when cells were exposed to the highest concentration of Emblica officinalis, Hemidesmus indicus, and Rubia cordifolia. All concentrations of Emblica officinalis tested inhibited the proliferation of osteogenic precursors, while only the highest doses of Asparagus racemosus and Rubia cordifolia were toxic. On the contrary, Hemidesmus indicus did not affect osteogenic precursor growth at any concentration tested.

Conclusion

Among the medicinal plants included in the study, Hemidesmus indicus showed the greatest antiosteoclastic activity without toxic effect on osteogenic precursors. Therefore, Hemidesmus indicus exhibits the properties of an antiresorptive drug and represents the ideal candidate for further clinical investigations.     

Enhancing oral moisture using an extract of Capparis masaikai Levl

The purpose of this study was to examine the effect of a Capparis masaikai Levl. extract on enhancing oral moisture. Solutions of Capparis masaikai extract, citric acid, sodium chloride, and sucrose were dropped on the tongue dorsum of 20 healthy subjects aged 23-34 years. After swallowing each solution, the oral moisture was measured for 60 min using a saliva wetness tester and a moisture checker. The subjects recorded the degree of taste using a visual analog scale to examine the stimulating effect of each solution on salivation. The Capparis masaikai extract had a long-lasting moistening effect on both the tongue dorsum and buccal mucosa for up to 60 min. The weakly bittersweet taste of the extract was perceived stronger than the other taste elements. The results suggest that the Capparis masaikai extract is useful for enhancing oral moisture.     

Hepatoprotective effect of Hypericum japonicum extract and its fractions

OBJECTIVE: To investigate the hepatoprotective activity of different parts of Hypericum japonicum against carbon tetrachloride(CCl(4))-induced hepatitis and alpha-naphthyl-isothiocyanate (ANIT)-induced cholestasis. MATERIALS AND METHODS: Mice were divided into groups and then administrated orally with solutions extracted from herbs before they were modeled in the experiments. Levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (T-BIL) in serum were evaluated. HPLC fingerprint was used for phytochemical analysis of the extracts. RESULTS: The total aqueous extract of Hypericum japonicum had an obvious effect on the decreasing of AST, ALT and T-BIL levels in serum. The isolated fraction IV (F4) exhibited a preferable activity of ameliorating cholestasis, while Fraction V (F5) was more efficacious in protecting liver from injury. Chemical fingerprint indicated that F5 contained several flavonoids which might be the active chemicals against hepatotoxicity. CONCLUSION: Different fractions of Hypericum japonicum manifest different effect, indicating their different potentials as candidacies of new drugs.