A Study on the Extraction Process of Active Ingredients from Akebia Stem and an Analysis of their Anti-Gastric Cancer Activity

The study investigated the extraction process of active ingredients from akebia stem and an analysis of their anti-gastric cancer activity. Three different extraction methods were used to obtain extracts, namely the decoction method (group A), reflux extraction method (group B), and maceration method (group C), of which reflux extraction method and maceration method used ethanol as the extraction solvent, while decoction method used distilled water for extraction. The differences in anti-gastric cancer activity of the three extracts were compared. MTT assay was used to test and compare the inhibitory effects of extracts obtained in A, B, and C groups on gastric cancer cells. The results showed that the dry extract obtained by heat reflux extraction with “water-ethanol” ratio of 1:2, extractant volume of 70 ml, with ethanol as extraction solvent presented the best inhibitory activity on gastric cancer SGC-7901 cells in this study. Its inhibitory effect did not change over time, and was directly proportional to the concentration.     

Study on Anti-Osteosarcoma Activity of Ethanol Extract of Venenum Bufonis In Vitro

Background

Venenum bufonis is the dried white secretion of the auricular and skin glands of Bufo gargarizans Cantor, or Bufo melanostictus Schneider, Bufonidae. It is used in the treatment of deep-rooted carbuncle, boils and swelling; pain in the throat, heart stroke, coma, abdominal pain, vomiting and diarrhea. The objective of this paper is to preliminarily observe the effects of ethanol extract of Venenum bufonis on growth, and proliferation of human osteosarcoma U2OS cell lines, and to provide a theoretical basis for an in-depth study of the clinical application of Venenum bufonis for osteosarcoma inhibition, with its mechanism of action.

Materials and Methods

SRB assay was used to determine the effect of Venenum bufonis ethanol extract on U2OS cell line activity, and to detect its inhibitory dose-effect on osteosarcoma cells. FCM was applied to determine the effect of Venenum bufonis ethanol extract on U2OS cell apoptosis and to perform cell cycle analysis.

Results

As results, different Venenum bufonis ethanol extracts showed apparent concentration-effect relationships on U2OS cell lines. FCM analysis showed that it had a U2OS apoptosis promoting effect, which increased with increasing concentration. Cell cycle analysis revealed that the Venenum bufonis ethanol extract mainly arrested U2OS in the G0/G1 phase, preventing the cells from progressing to the S phase.

Conclusion

The study concluded that Venenum bufonis ethanol extract has an inhibitory effect on proliferation of osteosarcoma U2OS cells.     

A Study on the Anticancer Activity of Ethanol Extract of Aristolochia Mollissima Hance on Osteosarcoma Hos Cells

This paper mainly studied the extraction process of traditional Chinese medicine Aristolochia mollissima Hance (Aristolochiaceae) and the inhibitory effect of its extracts on osteosarcoma HOS cells. The extraction process included the ultrasonic extraction method, heat reflux method and decoction method to obtain three different extracts. MTT assay was used to test the effect of the extracts on proliferation of HOS cells, to compare the degree of inhibitory activity of three extracts, and to calculate cell survival rates. The results showed that among the three extracts obtained by the ultrasonic extraction method, heat reflux method and decoction method, the one obtained by ultrasonic extraction has the largest yield, but the extract obtained by heat reflux method has the strongest anticancer activity. Nevertheless, the three extracts all have a good inhibitory activity on the proliferation of osteosarcoma HOS cells.     

Study on In Vitro Anti-Tumor Activity of Bidens Bipinnata L. Extract

We studied the in vitro anti-tumor activity of Bidens Bipinnata L. extract. MTT assay was used to investigate the inhibitory effect of different concentrations of the extracts on human hepatocellular carcinoma (HepG2) cell lines and human cervical carcinoma (Hela) cell lines, and the IC₅₀ values were calculated. The Bidens Bipinnata L. extract had different degrees of inhibitory effects on these two cells, and when exposure time was 48 h, the inhibition rate reached its peak, with IC₅₀ values of 14.80 µg/mL and 13.50 µg/mL respectively. The Bidens Bipinnata L. extract had a good inhibitory effect on human HepG2 cell lines and Hela cell lines, and thus has certain development prospects.     

A Study on the Effect of Ethanol Extract of Radix Rhapontici on Erythrocyte Immune Function in Rats

This paper mainly studied the effect of ethanol extract of Radix rhapontici on erythrocyte immune function in SD rats with acute blood stasis. The methods used the effect on erythrocyte immune function. After intragastric administration of suspension of ethanol extract of Radix rhapontici to SD rats for 3 weeks, on the 21st day from intragastric administration, SD rats were made into blood stasis model and bloods were collected to determine the C3b, C3bRR, RFIR, and RFER in erythrocyte immune function. Meanwhile, serum total antioxidant activity (TAA), superoxide dismutase (SOD) activity, and serum malondialdehyde (MDA) level of rats were determined, and experimental results were analysed with analysis of variance and Q test. The results showed that the ethanol extract of Radix rhapontici had a very good effect on enhancement of erythrocyte immune function in SD rats.     

Immunostimulating Effect of Aqueous Extract of Amphypterygium Adstringens on Immune Cellular Response in Immunosuppressed Mice

Amphypterygium adstringens is a Mexican tree known as cuachalalate whose bark is habitually used for the treatment of fresh wounds, gastric ulcers, gastrointestinal cancer and various inflammatory conditions. The aim of this study was to evaluate the immunostimulant effect of the aqueous extract of A. adstringens on immune cellular response in immunosuppressed mice. An aqueous extract from the bark of cuachalalate was administered into BALB/c mice for 10 days. We assessed their immunostimmulant activity on cellular immune response by Delayed Type Hypersensitivity Response (DHT) to dinitrofluorobencene (DNFB) and by MTT assay. L5178Y lymphoma was used as immunossuppression model. An increase in DHT was observed after treatment with 10 and 100 mg/kg of the aqueous extract from A. adstringens oral treatment in lymphoma bearing mice. Splenocyte proliferation rate was significantly increased (2.5 time) in immunosuppresed mice treated with 10 mg/kg oral treatment compared with group that received vehicle only. The present study showed for the first time the aqueous extract from A. adstringens as a positive immunostimulant agent in lymphoma bearing mice and we demonstrated evidence to support the traditionally use of cuachalalate in conditions in which the immune system is depressed.     

A Study on the Effect of Aqueous Extract of Lobelia Chinensis on Colon Precancerous Lesions in Rats

This paper studies the effects of Lobelia chinensis on colon precancerous lesions and on colonic epithelial proliferation and apoptosis in DMH-induced rats. After two weeks of feeding, 50 Wistar rats were randomly divided into five groups, namely the normal group, model group, Lobelia chinensis low-dose group, medium-dose group and high-dose group. Lobelia chinensis was made into ACF model, and administered to experimental groups for 10 consecutive weeks. Control group was given equivalent amount of normal saline. After feeding for 10 weeks, the rats in each group were sacrificed and the changes in colonic ACF number of rats in experimental groups were observed, and the inhibition rates were calculated. The results showed that among the rats fed for 24 h and 48 h, the number of apoptotic cells in colonic crypts of rats in DMH group did not differ significantly from the control group, while the difference was obvious between the control group and Lobelia chinensis treatment groups. The medium and high doses, that is, 0.45 g/kg and 1.35 g/kg can significantly inhibit ACF formation (P<0.01). The inhibition rates of low, medium and high doses were 8.12%, 59.42% and 65.44%, respectively.     

Study on the Antibacterial Activity of Bergenia Purpurascens Extract

Background

Bergenia purpurascens has tonic, haemostatic and anti-tussive actions. Anti-inflammatory and anti-bacterial activities of Bergenia purpurascens have not been reported so far. The objective of this paper is to provide experimental basis for the clinical application of Bergenia purpurascens through the pharmacodynamic study on its anti-inflammatory and anti-bacterial effects.

Methods

Experimental models of xylene-induced ear edema in mice, cotton pellet granuloma in rats, and acetic acid-induced peritoneal capillary permeability in mice were used to investigate the anti-inflammatory effect of Bergenia purpurascens; bacteriostatic and bactericidal effects of Bergenia purpurascens extract on Staphylococcus aureus (SA), methicillin-resistant Staphylococcus aureus (MRSA), and β-lactamase positive Staphylococcus aureus (ESBLs-SA), were observed in vitro.

Results

The results show that Bergenia purpurascens extract could markedly inhibit xylene-induced mouse ear edema, cotton pellet granulation tissue hyperplasia, and increased capillary permeability. Bergenia purpurascens extract has an inhibitory effect on SA, MRSA and ESBLs-SA.

Conclusion

We conclude that Bergenia purpurascens extract has certain anti-inflammatory and anti-bacterial effects.     

Antidiarrhoeal Activity of an Ethanol Extract of the Stem Bark of Piliostigma Reticulatum (Caesalpiniaceae) in Rats

Piliostigma reticulatum (Caesalpiniaceae) is used in Africa as a traditional medicine for the treatment of many diseases, such as malaria, tuberculosis and diarrhoea. We investigated the antidiarrhoeal properties of a crude ethanol extract from the stem bark of Piliostigma reticulatum (EEPR) in Wistar albino rats to substantiate its traditional use and to determine its phytochemical constituents. The antidiarrhoeal activity of the plant extract was evaluated in a castor oil-induced diarrhoea model in rats and compared with loperamide. The effect of the extract on gastrointestinal motility was also determined by the oral administration of charcoal meal and castor oil-induced intestinal fluid accumulation (enteropooling). EEPR showed remarkable dose-dependent antidiarrhoeal activity evidenced by a reduction of defecation frequency and change in consistency. Extracts at 250, 500 and 1000 mg/kg body weight significantly reduced diarrhoeal faeces. EEPR also significantly inhibited gastrointestinal motility and castor oil-induced enteropooling at 500 and 1000 mg/kg, similar to the inhibition obtained in control rats treated by atropine. Phytochemical screening revealed the presence of tannins, flavonoids, polyphenols and reducing sugars in the stem bark of P. reticulatum. No mortality or visible signs of general weakness were observed in the rats following administration of the crude extract in doses up to 6000 mg/kg body weight in an acute toxicity study. Our results show that the stem bark of P. reticulatum possesses antidiarrhoeal activity and strongly suggest that its use in traditional medicine practice could be justified.     

A Study on Anticancer Activity of Caulis Spatholobi Extract on Human Osteosarcoma Saos-2 Cells

The objective of the present study was to investigate the anticancer activity of Chinese medicine Caulis Spatholobi extract on multicentric osteosarcoma cells. Ultraviolet spectrophotometry was used to determine the total flavonoid content in each sample; vanillin sulphuric acid assay was used to determine the condensed tannin content in each sample; and the varying degrees of inhibitory activities of ethanol, ethyl acetate and n-butanol extracts of Caulis Spatholobi on human osteosarcoma Saos-2 cells were studied. The results showed that the inhibitory activity of ethyl acetate extract was the highest among the four extracts. The condensed tannin contents of 1.2 mg/mL Caulis Spatholobi water extract, ethanol extract, ethyl acetate extract and petroleum ether extract were 26.23%, 48.36%, 70.18% and 40.51% respectively; and condensed tannin content of 1.5 mg/mL Caulis Spatholobi water extract, ethanol extract, ethyl acetate extract and petroleum ether extract were 4.15%, 5.81%, 8.76% and 7.30% respectively.     

Study on In-Vivo Anti-Tumor Activity of Verbena Officinalis Extract

We investigated the anti-tumor effects of Verbena officinalis extract on H22 tumor-bearing mice and its effect on immune function. Mice model of H22 solid tumor was established, the mice were divided into five groups and administered the extract, later, tumors were removed and inhibition rates were calculated; spleens were removed and spleen indices were calculated, and the sheep red blood cell-delayed-type hypersensitivity (SRBC-DTH) and the serum hemolysin level were determined. The Verbena officinalis extract had anti-tumor effect, with the inhibition rate reaching 38.78%, it also increased the spleen index to a certain extent, in addition, the changes in DTA and HA were not obvious compared with the model group. The Verbena officinalis extract had in vivo anti-tumor effect, while causing no damage on the immune function.     

Protective Activity of the Stem Bark Aqueous Extract of Musanga Cecropioides in Carbon Tetrachloride- and Acetaminophen-induced Acute Hepatotoxicity in Rats

The hepatoprotective activities and the mechanisms of actions of Musanga cecropioides stem bark aqueous extract (MCW) were investigated on acute hepatocellular injuries induced by intraperitoneal (IP) carbon tetrachloride (CCl₄) (20% CCl₄/olive oil, 1.5 mL/kg) and 800 mg/kg/IP of acetaminophen (APAP) in normal saline, in male Wistar rats. Among the Yorubas (South-West Nigeria), cold decoction of MCW is used as a natural antidote for oral gastric poisonings, infective hepatitis and other liver diseases. Its hepatoprotective activities were monitored by assaying for the serum aminotransferases, ornithine carbamoyl transferase and the toxicant-induced histopathological lesions in rat livers 24 hours post-induction. These enzymes are markers of acute hepatic injuries and their elevations are indications of acute liver injuries. Pretreatment of rats with graded doses (125 – 500 mg/kg) of MCW significantly attenuated the acute elevation of the liver enzymes and the hepatotoxin-induced histopathological lesions in the rat livers. The presence of two active natural antioxidants (flavonoids and alkaloids) in high concentrations in MCW may account for the hepatoprotective activities observed in this study. These results, thus, support the folkloric use of MCW for treatment of hepatic injuries resulting from acute gastric poisonings, infective hepatitis or other liver diseases.     

Antiulcerogenic Activity of Ethanolic Leaf Extract of Lasianthera Africana

The effect of ethanolic leaf extract of Lasianthera africana on experimentally induced ulcer was studied in rats. The extract (1000 – 3000mg/kg) inhibited ethanol-induced, indomethacin - induced and reserpine -induced ulcer models in a dose dependent fashion. The various degrees of inhibitions were statistically significant (p<0.01). The effect of the extract was comparable to that of the standard drugs used. Thus, Lasianthera africana extract demonstrated a good antiulcer activity which supports the antiulcer effect of this plant in traditional medicine.     

Spasmogenic effect of the Aqueous Extract of Tamarindus Indica L. (Caesalpiniaceae) on the Contractile Activity of Guinea-Pig Taenia Coli

The effect of aqueous extract of Tamarindus indica (AETI) was studied on the guinea pig taenia coli, due to its use for treatment of constipation in traditional medicines. AETI, at concentrations ranging from 10⁻⁸ mg/ml to 10⁻² mg/ml, increased the spontaneous contractile activity of guinea pig taenia coli in a dose-dependent manner (EC50= 4×10⁻⁶ mg/ml). This activity was unaffected by atropine. In high K⁺, Ca²⁺-free solution containing EDTA, AETI as well as acetylcholine, used as a control, induced tonic contraction. These results suggest that the plant extract exert a spasmogenic effect that would not involve cholinergic mechanism of action. However, these active principles could mobilize both extra cellular calcium and intracellular calcium from internal stores.     

Chemopreventive Effect of Cousinia Shulabadensis Attar & Ghahraman Ethanol Extract

Matrix metalloprotainases (MMPs) play an important role in several pathologic processes such as malignancy in which they facilitate invasion and metastasis and can be targets for anticancer therapies. Here, in this study, we investigated the cytotoxicity effect of Cousinia shulabadensis Attar & Ghahraman extract as well as its impact on MMPs activity using a model of cell line (Fibrosarcoma-Wehi164). To assess anti-invasiveness potentials, a modified zymoanalysis method was used to measure MMP-2 and MMP-9 activities in the conditioned-media. The concentration necessary to produce 50% cell death was >80µg/ml for ethanol extract of Cousinia shulabadensis, while a 23 µg/ml concentration of the diclofenac sodium produced the same effect. The invasion of WEHI 164 cells was considerably inhibited at concentrations > 20 µg/ml by total plant extract. The total extract of the plant did not show high toxicity at all tested concentrations, but demonstrated significant inhibition of MMP activity in dose-response fashion.     

Anti-Nociceptive and Anti-Inflammatory Activities of Aqueous Leaves Extract of Ocimum Gratissimum (Labiate) in Rodents

The aqueous leaves extract of Ocimum gratissimum was investigated for anti-nociceptive and anti-inflammatory effects in mice and rats. The models used to study the effect on nociception are the acetic acid-induced abdominal constriction test, hot- plate method in mice. The anti-inflammatory effect was investigated employing the formalin-induced hind-paw oedema in rats. The extract caused a significant (p<0.05), dose-dependent inhibition of acetic acid-induced writhing and hot-plate method. The extract also exhibited anti-inflammatory effect which was significant (P<0.001) at all the three doses. The intraperitoneal LD₅₀ value of the extract was 1264.9mg/kg body weight in mice. Preliminary phytochemical screening revealed the presence of alkaloids, saponins, tannins and flavonoids. The results suggest the extract contained pharmacologically active principles, and supports the local application of the plant in painful and inflammatory conditions.     

Analgesic and Anti-Inflammatory Properties of Aqueous Extract from Leaves of Solanum Torvum (Solanaceae)

Solanum torvum is used in Cameroonian traditional medicine for the management of pain and inflammation. The present work assesses the pain-killing and anti-inflammatory properties of the aqueous extracts of Solanum torvum leaves. Acetic acid- and pressure- induced pains were reduced by this extract while carrageenan-induced inflammation was inhibited at various doses of the extract. The extract therefore has both analgesic and anti-inflammatory properties.     

Evaluation of Acute and Subacute Oral Toxicity of the Ethanol Extract from Antidesma Acidum Retz.

Toxicity tests of 95% ethanol extract of the root of Antidesma acidum were studied in male and female rats. The oral acute toxicity test at 5,000 mg/kg revealed that the ethanol extract did not produce toxic effects on signs, general behavious, mortality and gross appearance of internal organs of rats. Furthermore, the oral sub-acute toxicity test at the dose of 1,000 mg/kg/day displayed no significant changes in body and internal organs'' weights, normal hematological and clinical blood chemistry values. Histological examination also showed normal architecture of all internal organs. In conclusion, the ethanol extract of Antidesma acidum did not produce any toxicity in oral acute and suba-cute toxicity studies.     

Effect of Administration of Aqueous Extract of Hippobromus Pauciflorus Leaves in Male Wistar Rats

The effect of administration of aqueous extract of Hippobromus pauciflorus (L.f.) Radlk (Sapindaceae) leaves at 50, 100 and 200 mg/kg body weight for 14 days on some biochemical parameters in male Wistar rats was investigated. The extract at all the doses tested did not significantly (P>0.05) alter the levels of white blood cells, red blood cells, mean corpuscular volume, platelets, neutrophils, monocytes, lymphocytes and large unstained cells. While the levels of haemoglobin, packed cell volume and basophils increased significantly (P<0.05) at specific doses, the mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration and eosinophils decreased significantly (P<0.05). Again, the extract did not significantly (P<0.05) alter the computed liver- and kidney-body weight ratios, sodium, chloride and total protein, though, the levels of potassium, inorganic phosphorus, globulin, urea, total and conjugated bilirubin increased significantly (P<0.05) at certain doses. In contrast, the levels of albumin and creatinine also decreased significantly (P<0.05) at specific doses. While the activities of alkaline phosphatase, gamma glutamyl transferase and alanine aminotransferase remained significantly (P<0.05) unaltered in the serum, aspartate aminotransferase activity increased only at 200 mg/kg body weight. The atherogenic index as well as the concentrations of cholesterol, high- and low-density lipoprotein cholesterol in the serum of the animals were not significantly (P>0.05) altered. However, the extract significantly (P<0.05) increased the concentration of triacylglycerol. The results suggest that the extract has mild and dose specific haemato-, hepato- and nephrotoxic effects and may not be completely safe as oral remedy at the doses investigated.     

Anthelminthic Efficacy of Aqueous Extract of Acanthus Montanus Leaf Against Strongylid Nematodes of Small Ruminants

The anthelminthic efficacy of the crude aqueous extract of Acanthus montanus (Nees) T. Anders (Acanthaceae) against strongylid nematodes of small ruminants was investigated using the in-vitro egg hatch and larval growth inhibition assays. Faecal samples collected per rectum from sheep and goats were subjected to parasitological examination using the McMaster counting technique with a yield of 700 eggs per gram (E.P.G.) of faeces. Crude aqueous leaf extract of Acanthus montanus was extracted using cold water extraction with a yield of 13.01% w/w. Egg hatch assay revealed a 91.75% reduction in egg hatch at concentration of 25mg/ml of the extract. The extract had a 100% inhibition, at 200mg/ml concentration which was equivalent to the activity of 3.125mg/ml albendazole. The distilled water control however, showed a 0% inhibition. Larval growth inhibition assay results showed the extract on Day 1 at 25mg/ml and 200mg/ml concentrations yielded 67.02% and 85.26% larval inhibition. On Day 2, 100% inhibition was obtained on all concentrations of the extract except for 25mg/ml that yielded 88.30%. Albendazole however, had a 100% larval inhibition for all concentrations on Day 2. A 100% larval inhibition was recorded on Day 3 from the 25mg/ml concentrations. The mean percentage larval inhibition of the extract at 200mg/ml (92.63%) was comparable to the standard anthelminthic (albendazole) at 12.50mg/ml (92.28%). These findings showed that there is a pharmacological basis for the folkloric claim of the anthelminthic effect of Acanthus montanus.