Hempseed oil induces reactive oxygen species- and C/EBP homologous protein-mediated apoptosis in MH7A human rheumatoid arthritis fibroblast-like synovial cells

Ethnopharmacological relevance

The medicinal efficacy of hempseed (Cannabis sativa L.), which is rich in polyunsaturated fatty acids, in atopic dermatitis, inflammation, and rheumatoid arthritis (RA) has been suggested for centuries. Hempseed has been used as a treatment for these diseases in Korean and Chinese folk medicine. The aim of the study is to investigate the effects of hempseed oil (HO) on MH7A human RA fibroblast-like synovial cells.

Materials and methods

MH7A cells were used to study the anti-rheumatoid effects of hempseed (Cannabis sativa L., cv. Cheungsam/Cannabaceae) oil by investigating cell viability, apoptosis, lipid accumulation, oxidative stress, and endoplasmic reticulum (ER) stress-induced apoptosis.

Results

HO treatment reduced the survival rate of MH7A cells and promoted apoptotic cell death in a time- and dose-dependent manner. Both lipid accumulation and the level of intracellular reactive oxygen species (ROS) increased in HO-treated MH7A cells. Co-treatment with the antioxidant Tiron effectively abrogated the cytotoxic effects of HO; the ROS level was reduced, cell viability was recovered, and apoptotic cell death was significantly diminished. Moreover, HO-treated cells exhibited increased expression of the major ER stress markers, glucose-regulated protein 78 and C/EBP homologous protein (CHOP). The siRNA-mediated knockdown of CHOP prevented HO-induced apoptosis.

Conclusions

Our results suggest that HO treatment induced lipid accumulation, ROS production, CHOP expression, and apoptosis in MH7A cells, and that CHOP functions as an anti-rheumatoid factor downstream of HO in MH7A cells.     

Interactions of pharmacokinetic profile of different parts from Ginkgo biloba extract in rats

Ethnopharmacological relevance

Extracts from Ginkgo biloba L. leaves confer their therapeutic effects through the synergistic actions of flavonoid and terpenoid components, but some non-flavonoid and non-terpenoid components also exist in this extract. In the study of this paper, an investigation was carried out to compare the pharmacokinetic parameters of fourteen compounds to clarify the influences of non-flavonoid and non-terpenoid fraction (WEF) on the pharmacokinetics profile of the flavonoid fraction (FF) and the terpene lactone fraction (TLF) from Ginkgo biloba extracts.

Materials and methods

A selective and sensitive UPLC–MS/MS method was established to determine the plasma concentrations of the fourteen compounds to compare the pharmacokinetic parameters after orally administration of FF, TLF, FF–WEF, FF–TLF, TLF–WEF and FF–TLF–WEF with approximately the same dose. At different time points, the concentration of rutin (1), isoquercitrin (2), quercetin 3-O-[4-O-(-β-D-glucosyl)-α-L-rhamnoside] (3), ginkgolide C (4), bilobalide (5), quercitrin (6), ginkgolide B (7), ginkgolide A (8), luteolin (9), quercetin (10), apigenin (11), kaempferol (12), isorhamnetin (13), genkwanin (14) in rat plasma were determined and main pharmacokinetic parameters including T_1/2, T_max, C_max and AUC were calculated using the DAS 3.2 software package. The statistical analysis was performed using the Student?s t-test with P<0.05 as the level of significance.

Results

FF and WEF had no effect on the pharmacokinetic behaviors and parameters of the four terpene lactones, but the pharmacokinetic profiles and parameters of flavonoids changed while co-administered with non-flavonoid components. It was found that C_max and AUC of six flavonoid aglycones in group FF–WEF, FF–TLF and FF–TLF–WEF had varying degrees of reduction in comparison with group FF, especially in group FF–TLF–WEF. On the contrary, the values of C_max, T_max and AUC of four flavonoid glycosides in group FF–TLF–WEF were significantly increased compared with those in group FF.

Conclusions

These results indicate that non-flavonoid components in Ginkgo biloba extracts could increase the absorption and improve the bioavailability of flavonoid glycosides but decrease the absorption and reduce the bioavailability of flavonoid aglycones.     

Acute and sub-chronic toxicity profile of methanol leaf extract of Gouania longipetala in rats

Ethnopharmacological relevance

Gouania longipetala leaves are commonly used in folkloric medicine in Africa and other parts of the world for treatment of edema, febrifuges, veneral diseases, lumbago, heart diseases, diabetes mellitus malaria, etc. This study therefore evaluated safety profile of the methanol leaf extract of the plant using acute and sub-chronic studies in rat model.

Materials and methods

Acute toxicity test of the plant lasted for 48 h with oral administration of graded doses (100–4000 mg/kg) of Gouania longipetala extract (GLE) in rats. The rats were observed for signs of toxicity and death. The sub-chronic toxicity was evaluated by administration of different doses (2.5, 5 and 10 mg/kg) of GLE daily in feed for 90 days. On days, 30, 60 and 90, blood samples collected from the retro-orbital plexus of the eye of the rats were used for evaluation of serum biochemistry, hematology, lipid peroxidation and in vivo antioxidant activities. Histopathological evaluations of the kidney, liver, lungs and heart were also done.

Results

The acute toxicity test revealed no observable signs of toxicity or morbidity. Sub-chronic study showed that GLE significantly (p<0.05) increased relative liver weight on day 90 at 10 mg/kg. There were no significant variations in the hematological parameters of both GLE treated and untreated rats. The extract significantly (p<0.05) reduced total cholesterol, triglycerides, very low density lipoproteins and increased high density lipoproteins which was more prominent on day 90 at the dose of 10 mg/kg. The extract significantly (p<0.05) increased liver enzyme markers at the doses used. GLE also significantly (p<0.05) increased serum urea at the dose of 10 mg/kg on day 90. The extract caused dose-dependent and significant (p<0.05) increase in superoxide dismutase and decrease in malondiadehyde. Histopathological studies revealed degenerative changes in the kidney and liver.

Conclusion

The results of the study suggest that Gouania longipetala is well tolerated in short term therapies, but may have long term toxic effects on the kidney and liver.     

Effect of Bridelia ferruginea Benth (Euphorbiaceae) ethyl acetate and acetone fractions on insulin resistance in fructose drinking mice

Ethnopharmacological relevance

The root of Bridelia ferruginea is traditionally used as a treatment for type 2 diabetes. The present study was investigated to evaluate the effect of Bridelia ferruginea root bark fractions on some markers of type 2 diabetes on fructose drinking mice.

Materials and methods

Mice received a solution of fructose 15% during 42 days ad libitum; at the 15th day to the 42nd day, they received distilled water for fructose drinking control group, metformin 50 mg/kg per day or fractions 50 mg/kg per day for treatment groups. The normal control group received only distilled water during the experiment. After 6 weeks of experiment, OGTT, fasting blood glucose, plasma insulin, triglycerides (TG), total cholesterol, AST and ALT levels were measured.

Results

Fructose drinking control group (F) showed significant (p<0.001) increase of glucose tolerance, plasma levels of total cholesterol, triglycerides and insulin index for insulin resistance (Homeostasis Model Assessment ratio HOMA-IR) as compared to normal control mice. In treated groups, there was a significant reduction of glucose intolerance respectively 74% (p<0.001), 25% (p<0.5) and 92% (p<0.001) for ethyl acetate fraction, acetone fraction and metformin at the same dose of 50 mg/kg per day during 4 weeks administration. In ethyl acetate fraction and metformin treated groups, biochemical parameters and insulin index were significantly (p<0.001) lower than that of fructose drinking control group.

Conclusions

This indicates that Bridelia ferruginea root bark ethyl acetate fraction improved insulin resistance as metformin significantly in type 2 diabetes.     

Plasma pharmacokinetics and lung distribution of tetrahydropalmatine after topical application of cold asthma recipe extract: Feishu (BL 13) versus Non-Feishu acupoint

Ethnopharmacological relevance

Acupoint application of cold asthma recipe (CAR) was a traditional Chinese medicine (TCM) method, widely used as an alternative medicine for clinical prevention of the common winter diseases of asthma and bronchitis. Tetrahydropalmatine (THP) was a main active ingredient of CAR extract. The aim of this study is to compare plasma pharmacokinetics and lung distribution of THP between Feishu (FS) acupoint (BL 13) and Non-Feishu (NFS) acupoint application of CAR extract by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).

Materials and methods

The extract of CAR was topically administrated in FS and NFS acupoint of rats for plasma pharmacokinetics, and topically administrated in FS and NFS acupoint of mice for lung distribution. The plasma and lung homogenates were pretreated by protein precipitation with acetonitrile. Chromatographic separation was performed on an ACQUITY UPLC BEH C18 column with a mobile phase consisted of 0.1% formic acid in acetonitrile and 0.1% formic acid in water. The detection was accomplished by multiple-reaction monitoring (MRM) scanning in the positive electrospray ionization (ESI⁺) mode. All pharmacokinetic parameters were estimated by non-compartmental analysis.

Results

A sensitive, accurate and precise UPLC-MS/MS method was successfully established for determination of THP in 100 μL plasma and lung homogenate. The lower limit of quantification (LLOQ) of THP was 0.05 ng/mL and 0.072 ng/mL, respectively. The pharmacokinetic results manifested that THP was absorbed and eliminated slowly in plasma. Additionally, it was found that there was significantly higher amount of THP absorbed into blood and lung after FS acupoint application compared to NFS acupoint application.

Conclusions

Both of the rat plasma pharmacokinetics and mice lung distribution of THP could support that FS acupoint application of CAR extract has greater advantages of absorption into the blood circulation and distribution in target tissue over NFS acupoint application. The results might be helpful in providing a rational explanation for why the TCM chose the acupoint application and elucidating the underlying mechanism of this treatment.     

Effects of hydroxysafflor yellow A on the activity and mRNA expression of four CYP isozymes in rats

Ethnopharmacological relevance

In traditional therapy with Chinese medicine, hydroxysafflor yellow A (HSYA), a main active component isolated from the dried flower of Carthamus tinctorius L., is the principal efficiency ingredient of Safflor Yellow Injection. Now HSYA has been demonstrated to have good pharmacological activities of antioxidation, myocardial and cerebral protective and neuroprotective effects. The purpose of this study was to find out whether HSYA influences the effect on rat cytochrome P450 (CYP) enzymes (CYP1A2, CYP2C11, CYP2D4 and CYP3A1) by using cocktail probe drugs in vivo; the influence on the levels of CYP mRNA was also studied.

Materials and methods

A cocktail solution at a dose of 5 mL/kg, which contained phenacetin (20 mg/kg), tolbutamide (5 mg/kg), dextromethorphan (20 mg/kg) and midazolam (10 mg/kg), was given as oral administration to rats treated with short or long period of intravenous HSYA via the caudal vein. Blood samples were collected at a series of time-points and the concentrations of probe drugs in plasma were determined by HPLC–MS/MS. The corresponding pharmacokinetic parameters were calculated by the software of DAS 2.0. In addition, real-time RT-PCR was performed to determine the effect of HSYA on the mRNA expression of CYP1A2, CYP2C11, CYP2D4 and CYP3A1 in rat liver.

Results

HSYA had significant inhibition effects on CYP1A2 and CYP2C11 in rats as oriented from the pharmacokinetic profiles of the probe drugs. Furthermore, HSYA had no effects on rat CYP2D4. However, CYP3A1 enzyme activity was induced by HSYA. The mRNA expression results were in accordance with the pharmacokinetic results.

Conclusions

HSYA can either inhibit or induce activities of CYP1A2, CYP2C11 and CYP3A1. Therefore, co-administration of some CYP substrates with HSYA may need dose adjustment to avoid an undesirable herb–drug interaction.     

New finding of an anti-TB compound in the genus Marsypopetalum (Annonaceae) from a traditional herbal remedy of Laos

Ethnopharmacological relevance

There is widespread use of traditional herbal remedies in the Lao PDR (Laos). It is common practice to treat many diseases with local plants. This research project documented and analysed some of these traditional remedies used to treat symptoms of tuberculosis (TB).

Materials and methods

This research was executed by interviewing healers about plants used traditionally to treat the symptoms of TB. Samples of some of the plants were collected, and extracts of 77 species were submitted to various in vitro assays in order to determine the amount of growth inhibition of virulent Mycobacterium tuberculosis H37Rv (Mtb), as opposed to other microbes and mammalian Vero cells.

Results

Interviews took place with 58 contemporary healers in 5 different provinces about plants currently used, giving a list of 341 plants. Bioassay-guided fractionation was performed on Marsypopetalum modestum (Pierre) B. Xue and R.M.K. Saunders (Annonaceae), leading to the isolation of dipyrithione, an anti-mycobacterial compound isolated for the first time from the genus Marsypopetalum through this research.

Conclusions

This research has helped to increase awareness of Laos’ rich diversity of medicinal plants and will hopefully provide incentive to preserve the undeveloped forested areas that remain, which still hold a wealth of medical information for future discoveries.     

Confocal mirco-Raman spectroscopic analysis of the antioxidant protection mechanism of the oligosaccharides extracted from Morinda officinalis on human sperm DNA

Ethnopharmacological relevence

Male infertility is a stressful and frustrating problem for the society, but a number of male infertility treatments are available as traditional Chinese medicine strategies which have been tried with variable success, while evidence is still limited on whether-or how much-herbs or supplements might help increase fertility, so the aim of this study was to investigate if the oligosaccharides extracted from Morinda officialis, a Chinese herb, is the active constituents to the fertility.

Materials and methods

In this study, we prepared the H₂O₂-demaged human sperm, cocultured with the oligosaccharides in vitro, then observed the changes of the DNA using confocal micro-Raman spectroscopy, and comparative analysis the differences of the spectra of different treated groups.

Results

The results showed that the oligosaccharides extracted from Morinda officialis can keep the “Raman fingerprints” of the human sperm DNA almost the same as those of the control groups, but very different from the H₂O₂-induced groups, especially the intensity of bands at 787, 993, 1094, 1254, 1340, 1376, 1421, 1443, 1487, 1577 and 1662 cm⁻¹ which could be as potential targets for the drugs finding, and further principal component analysis was successfully used to classify the Raman spectra of normal control and model groups.

Conclusion

This results suggested that the oligosaccharides can protect the DNA of human sperm from being damaged by H₂O₂, and which was one of the active constituents of Morinda officialis on treating infertility. It was also demonstrated that Morinda officialis as a tonifying and replenishing natural herb medicine can be used to enhance reproductive functions, and the Raman spectroscopy could be an applicable technology for screening active components in vitro from herbs.     

Arnebin-1 promotes the angiogenesis of human umbilical vein endothelial cells and accelerates the wound healing process in diabetic rats

Ethnopharmacological relevance

Zicao is a traditional wound healing herbal medicine that has been used for several hundred years in China. A survey of the published literatures revealed that arnebin-1, one of the naphthoquinone derivatives, played the most important role in wound healing property of this plant. However, whether arnebin-1 affects angiogenesis in vitro and has an effect on wound healing process in diabetic rats remains enigmatic. To investigate the effect of arnebin-1 with or without VEGF on proliferation, migration and tube formation of HUVECs in vitro and the effect of its topical application in the form of ointment on wound healing in a cutaneous punch wound model of alloxan-induced diabetic rats in vivo.

Materials and methods

The pro-angiogenic functions of arnebin-1 on HUVECs including proliferation, migration and angiogenesis were evaluated through MTT assay, wound healing assay, transwell assay and tube formation assay in vitro. Male Sprague-Dawley rats were injected intraperitoneally with alloxan to induce type ? diabetic rats. Three wounds were created in each rat on the dorsal surface, and then divided to be basement treated, arnebin-1 ointment treated and untreated group correspondingly. The indicators including wound closure rate and histological evaluation were investigated on day 4 and 7 post-wounding.

Results

Without VEGF, arnebin-1 did not affect the proliferation of HUVECs significantly, but had a positive effect on cell migration and tube formation. However, in the presence of minimal VEGF, Arnebin-1 could increase the proliferation, enhance the migration and promote the tube formation of HUVECs significantly. The wound closure rate was increased significantly in arnebin-1 treated group compared to that of untreated and basement treated groups in diabetic rats, and the histological evaluation also showed well organized dermal layer, reduced number of macrophages, increased number of fibroblasts, remarkable degree of neovascularization and epithelization in arnebin-1 treated group.

Conclusion

These findings suggest that arnebin-1 has a pro-angiogenic effect, and a synergetic effect with VEGF promotes the wound healing process in diabetic rats.     

Ethnobotanical study of medicinal plants of Sirjan in Kerman Province,Iran

Ethnopharmacological relevance

From ancient time human beings have used different plants, animals and minerals to prevent and treat various diseases. In this respect, plants have been of particular importance. Ethnobotany is the science of reviewing how indigenous people and local tribes have used their regional plants for particular purposes such as treating diseases in the past. The information gathered from such studies can help to improve national health systems and even lead to the discovery of new medicines. Keeping this in mind, in this study a survey of Sirjan area ethnobotany was conducted during 2011–2012.

Materials and methods

At the beginning, thirteen local people were interviewed about their region?s medicinal plants and their consumptions. These plants were collected and identified using identification keys. The data collected was analyzed using quantitative value indices F_IC, RFC and CI.

Result

Many plants collected have medicinal properties and have been used by local people to treat various ailments. Of these plants, nineteen families, 37 genera and 43 species belonged to medicinal plants. Among them, Lamiaceae with 8 species and Malva L. with 3 species were the largest medicinal plant families and genera, respectively. These plants are often used as decoction (28%) and as powder (21%). Also, the fruit of these plants are used most often. Besides being used as medicinal plants, they have other uses such as food, fuel, etc. Malva sylvestris has the largest value of relative frequency of citation and cultural importance indices. The most ailment categories have the highest level of informant agreement (mean F_IC=0.92).

Conclusion

Despite the semi-desert climate and lack of rich vegetation, many medicinal and economic plants are found in Sirjan region. Uncontrolled harvesting of the medicinal plants such as Bunium persicum, Cuminum cyminum, Zataria multiflora and Satureja bachtiarica in this region by local people has increased the risk of their extinction and calls for a restrict control over their protection by the authorities.     

Inhibition of the DHT-induced PSA secretion by Verbascum xanthophoeniceum and Serenoa repens extracts in human LNCaP prostate epithelial cells

Ethnopharmacological relevance

Verbascum xanthophoeniceum is a mullein plant, typical of Balkan region and some parts of Turkey, traditionally used as phytotherapeutic agent due to its anti-inflammatory properties. It is rich in phenylethanoid and iridoid metabolites whose anti-inflammatory properties are under characterization.

Materials and methods

The role of Verbascum xanthophoeniceum crude methanolic extract and its isolated phenylethanoid glycoside verbascoside have been evaluated, in comparison to a saw palmetto extract, on a human in vitro model of androgen-regulated prostate epithelium, the LNCaP cell line. Cytotoxicity and DHT-induced free and total PSA secretion have been thoroughly studied.

Results

We have found that similar to saw palmetto, Verbascum xanthophoeniceum extract and its isolated phenylethanoid glycoside verbascoside have no cytotoxicity in human LNCaP prostate epithelial cells, whereas an inhibitory effect on the DHT-induced free and total PSA secretion, a recognized anti-androgen like activity, has been shown in case of both Verbascum xanthophoeniceum extract and pure verbascoside. Furthermore, in the absence of the endogenous androgen DHT, an androgen-like activity in Verbascum xanthophoeniceum is detectable as it is for saw palmetto, suggesting that a mixed androgen-antiandrogen activity is present.

Conclusions

For the first time, Serenoa repens and Verbascum xanthophoeniceum extracts have shown an absence of cytotoxicity and an inhibitory effect on DHT-induced PSA secretion in an in vitro model of human prostate epithelium, whereas the phenylethanoid glycoside verbascoside appeared to explain only part of the Verbascum xanthophoeniceum inhibitory activity on PSA secretion.     

Cyto-/genotoxic effects of the ethanol extract of Chan Su,a traditional Chinese medicine,in human cancer cell lines

Ethnopharmacological relevance

Chan Su, an ethanolic extract from skin and parotid venom glands of the Bufo bufo gargarizans Cantor, is widely used as a traditional Chinese medicine for cancer therapy. Although the anti-cancer properties of Chan Su have been investigated, no information exists regarding whether Chan Su has genotoxic effects in cancer cells. The aim of the present study was to examine the cyto-/genotoxic effect of Chan Su in human breast carcinoma (MCF-7 cells), human lung carcinoma (A-549 cells), human T cell leukemia (Jurkat T cells), and normal human lymphocytes.

Materials and methods

Effects on the viability of MCF-7, A-549, Jurkat T cells, and normal lymphocytes were evaluated by Trypan blue exclusion assays. The DNA content in the sub-G1 region was detected by propidium iodide (PI) staining and flow cytometry. The genotoxicity of Chan Su was assessed by single-cell gel electrophoresis (comet assay) and the cytokinesis-block micronucleus assay (CBMN assay).

Results

Chan Su significantly inhibited the viability of MCF-7, A-549, and Jurkat T cells dose dependently, but had no effect on normal human lymphocytes. Apoptotic death of the cancer cells was evident after treatment. Chan Su also induced genotoxicity in a dose-dependent manner, as indicated by the comet and cytokinesis-block micronucleus assays.

Conclusions

These findings suggest that Chan Su can induce apoptotic death of, and exert genotoxic effects on, MCF-7, A-549, and Jurkat T cells.     

Antigenotoxic and antioxidant effects of the Mongolian medicinal plant Leptopyrum fumarioides (L): An in vitro study

Ethnopharmacological relevance

Leptopyrum fumarioides has been used in the traditional medicine of Mongolia for the treatment of various diseases, including drug intoxications. However, since there is only sparse information about its chemistry, active components, and pharmacological and toxicological effects, the major aim of the present study employing mouse lymphoma cells was to evaluate the genotoxic and antigenotoxic/antioxidative effects of extracts and components isolated from this plant.

Material and methods

A crude methanol extract was separated into three different sub-extracts: dichloromethane, n-butanol, and water. The major constituent of the n-butanol extract, i.e., the flavone luteolin-7-O-glucoside and a mixture of the most abundant compounds in the dichloromethane sub-extract were then isolated. DNA damage was evaluated using the comet assay; the antioxidant activity was evaluated using the DPPH radical scavenging assay.

Results

The crude methanol extract, the dichloromethane sub-extract and the mixture of compounds isolated from the latter fraction, increased the level of DNA damage after three hours of exposure. In contrast, no increase in DNA damage was observed in the cells that had been exposed to the n-butanol and water sub-extracts, or to the pure flavone. When non-DNA damaging concentrations of extracts and compounds were tested together with the DNA damaging agent catechol, all sub-extracts were found to reduce the catechol-induced DNA damage (the flavone was then found to be the most effective protective agent). The n-butanol sub-extract and the flavone were also found to have the most prominent antioxidative effects.

Conclusion

Based on the results from the present study, components in Leptopyrum fumarioides were found to protect the DNA damage induced by catechol, probably by acting as potent antioxidants.     

玛咖质量标准研究

为了建立玛咖定性、定量标准,参照2010年版药典附录相关方法,分别对玛咖的水分、浸出物、总灰分、酸不溶成分、重金属含量进行了测定;以玛咖中特征性酰胺类成分亚油酰胺为指标性成分,分别采用TLC和HPLC进行定性鉴别和含量测定研究。结果显示,玛咖中亚油酰胺的薄层色谱鉴别特征明显,专属性强;含量测定线性范围为0.01~2.00 μg(r=0.999 8),平均回收率(n=9)为99.27%(RSD 2.0%) 。所建立的玛咖定性鉴别和定量测定方法简单准确,能够为玛咖的质量控制提供有效数据。     

Role of human gut microbiota metabolism in the anti-inflammatory effect of traditionally used ellagitannin-rich plant materials

Ethnopharmacological relevance

Ellagitannin-rich plant materials are widely used in traditional medicine as effective, internally used anti-inflammatory agents. Due to the not well-established bioavailability of ellagitannins, the mechanisms of observed therapeutic effects following oral administration still remain unclear. The aim of the study was to evaluate if selected ellagitannin-rich plant materials could be the source of bioavailable gut microbiota metabolites, i.e. urolithins, together with determination of the anti-inflammatory activity of the metabolites produced on the THP-1 cell line derived macrophages model.

Materials and Methods

The formation of urolithins was determined by ex vivo incubation of human fecal samples with aqueous extracts from selected plant materials. The anti-inflammatory activity study of metabolites was determined on PMA differentiated, IFN-γ and LPS stimulated, human THP-1 cell line-derived macrophages.

Results

The formation of urolithin A, B and C by human gut microbiota was established for aqueous extracts from Filipendula ulmaria (L.) Maxim. herb (Ph. Eur.), Geranium pratense L. herb, Geranium robertianum L. herb, Geum urbanum L. root and rhizome, Lythrum salicaria L. herb (Ph. Eur.), Potentilla anserina L. herb, Potentilla erecta (L.) Raeusch rhizome (Ph. Eur.), Quercus robur L. bark (Ph. Eur.), Rubus idaeus L. leaf, Rubus fruticosus L. and pure ellagitannin vescalagin. Significant inhibition of TNF-α production was determined for all urolithins, while for the most potent urolithin A inhibition was observed at nanomolar concentrations (at 0.625 μM 29.2±6.4% of inhibition). Urolithin C was the only compound inhibiting IL-6 production (at 0.625 μM 13.9±2.2% of inhibition).

Conclusions

The data obtained clearly indicate that in the case of peroral use of the examined ellagitannin-rich plant materials the bioactivity of gut microbiota metabolites, i.e. urolithins, has to be taken under consideration.     

魟鱼软骨多糖对Lewis肺癌MMP-9表达的影响

目的:鱼软骨多糖(RCG)对Lewis肺癌基质金属蛋白酶9(MMP-9)表达的影响。方法:建立小鼠Lewis肺癌模型,将动物分为生理盐水组、不同剂量鱼软骨多糖组、环磷酰胺组,观察肿瘤生长情况,绘制肿瘤生长曲线,计算原发瘤抑制率;采用RT-PCR和Western blot检测瘤组织MMP-9基因和蛋白表达。结果:该多糖各剂量组小鼠的肿瘤生长曲线平缓,小鼠原发瘤抑制率、肺转移数与生理盐水组比较差异有显著性(P<0.05;P<0.01);与生理盐水比较,鱼软骨多糖各剂量组,MMP-9 mRNA和蛋白表达水平显著降低(P<0.01)。结论:鱼软骨多糖明显抑制小鼠Lewis肺癌原发瘤的生长和转移,可能与其抑制肿瘤MMP-9基因和蛋白表达有关。     

Antihypertensive effects of extract from Picrasma quassiodes (D. Don) Benn. in spontaneously hypertensive rats

Ethnopharmacologica relevance

Picrasma quassiodes (D. Don) Benn. (PQB) is a widely used herbal medicine used for gastroenteritis, snakebite, infection and hypertension in China. The aim of the study was to investigate the possible antihypertensive mechanisms on spontaneously hypertensive rats (SHR) of the extract from Picrasma quassiodes (D. Don) Benn.

Materials and methods

In the in vivo study, extract from Picrasma quassiodes (D. Don) Benn. at the dose of 50, 100, 200 mg/kg and captopril (12.5 mg/kg) were administrated to different group of SHR rats by gavage for six consecutive weeks after the blood pressures were firstly measured. At the end of the study, rats serum nitric oxide (NO) was measured by the nitrate reductase method; superoxide dismutase (SOD) and malondialdehyde (MDA) activities were measured by the colorimetric method; the expression of aorta endothelial nitric oxide synthase (eNOS) was measured by immunohistochemical analysis.

Results

The results showed that the oral administration of PQB could lower the systolic blood pressure (SBP) of SHR rats. In addition, the serum level of NO in SHR treated with PQB (100 and 200 mg/kg) was increased dramatically (P<0.05, P<0.01), but administration with captopril had no significant effect. The expression of aorta eNOS was markedly increased when treated with PQB. The serum SOD levels were increased with treatment of PQB (100 and 200 mg/kg; P<0.05, P<0.01). All the effects of these parameters were comparable to that of the SHR control group.

Conclusions

Our results disclosed that PQB is effective to lower blood pressure of SHR, its antihypertensive effect is probably associated with lowering oxidative stress by reducing SOD activity, preserving endothelial function and increasing the expression of eNOS to regulate NO and directly relax artery.     

Immuno-enhancement effects of ethanol extract from Cyrtomium macrophyllum (Makino) Tagawa on cyclophosphamide-induced immunosuppression in BALB/c mice

Ethnopharmacological relevance

Cyrtomium macrophyllum (Makino) Tagawa has been traditionally used as a herbal medicine for the treatment of various infectious diseases such as tapeworm infestation, colds, and viral diseases. However, no systematic study of the immunity of Cyrtomium macrophyllum ethanol extracts (CM) has yet been reported. The present work evaluates these traits.

Materials and methods

120 male BALB/c mice were divided into 6 groups of 20 mice each: (1) normal group (sterile physiological saline), which served as a blank control; (2) model group (Cyclophosphamide, CY) group (sterile physiological saline), which served as a negative control; (3) low-dose CM (50 mg/kg BW); (4) intermediate-dose CM (100 mg/kg BW); (5) high-dose CM (200 mg/kg BW); (6) CM group (200 mg/kg BW). CY (0.2 ml) was administered via intraperitoneal injection. The other regimens were administered via gavage in 0.2 ml solution. Phytochemical of CM was characterized by HPLC–LTQ-Orbitrap. The acute toxicity effect of the ethanol extract of Cyrtomium macrophyllum was also investigated.

Results

The spleen and thymus indices of mice receiving low, intermediate, and high doses of CM recovered more quickly than those of CY mice, and they did so in a dose-dependent manner. These mice also showed higher T cell and B cell proliferation responses and macrophage function than those of CY mice, and their serum levels of interleukin-6 and interferon-γ had become normal. In acute toxicity test, CM exhibited no mortality and behavioral changes in mice. Quantitative phytochemical analysis showed flavonoids, polyphenols, and tannins to be the major compounds present in the extract, at 27.64%, 30.87%, and 11.22%, respectively. We found that 16 compounds were characterized by the interpretation of their mass spectra obtained by the MS/MS.

Conclusion

The current study demonstrates that Cyrtomium macrophyllum ethanol extract improved immune function in CY-treated mice.