Anti-inflammatory activity of Chrysanthemum indicum extract in acute and chronic cutaneous inflammation

Aim of the study

Although Chrysanthemum indicum Linné (Compositae) has long been used in traditional Korean, Chinese, Japanese medicine to treat various immune-related diseases the underlying mechanism(s) by which these effects are induced remains to be defined in vivo model system. We investigated the effects of 70% ethanolic extract from Chrysanthemum indicum Linné (CIE) on skin inflammation in mice.

Materials and methods

Production of pro-inflammatory cytokines (TNF-α and IL-1β), activation of myeloperoxidase, and histological assessment were examined in acute and chronic skin inflammation using 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced mouse ear edema.

Results

CIE inhibited topical edema in the mouse ear, following administration at 200 mg/kg (i.p.), leading to substantial reductions in skin thickness and tissue weight, inflammatory cytokine production, neutrophil-mediated myeloperoxidase activity, and various histopathological indicators. Furthermore, CIE was effective at reducing inflammatory damage induced by chronic TPA exposure.

Conclusions

These results demonstrate that CIE is an effective anti-inflammatory agent in murine phorbol ester-induced dermatitis, and suggest that the extract may have therapeutic potential in a variety of immune-related cutaneous diseases.     

In vitro and in vivo toxicological evaluation of extract and fractions from Baccharis trimera with anti-inflammatory activity

Ethnopharmacological relevance

Baccharis trimera (Less) DC. (Asteraceae), popularly known in Brazil as “carqueja”, have been used in folk medicine to treat gastrointestinal, hepatic and renal diseases, and inflammatory processes as rheumatism.

Aim of the study

To evaluate the in vitro and in vivo toxicological effects of anti-inflammatory Baccharis trimera aqueous extract and fractions.

Materials and methods

Aqueous extract of Baccharis trimera (AEBt) was produced by infusion in boiling water. After lyophylization AEBt was extracted with 80% ethanol, originating the ethanolic supernatant fraction (EFBt) and the aqueous sediment fraction (AFBt). Anti-inflammatory properties of AEBt, EFBt or AFBt (3, 30 or 300 μg/kg b.w.) were evaluated by the carrageenan-induced mouse paw edema using indomethacin (10 mg/kg) as positive control. The growth of rat hepatoma cells (HTC) and human embryo kidney epithelial cells (HEK) was determined by protein staining assay. Cytotoxicity was assayed by the tetrazolium salt (MTT) reduction. Cyclosporin was used as reference cytotoxic drug for spleen cells and doxorubicin for HTC and HEK cells. For in vivo toxicological evaluation SW male mice were daily and oral (gavage) treated with extract/fractions at 4.2 mg/kg or 42 mg/kg during 15 days. After treatment liver or kidney cells were submitted to comet assay to determine the DNA damage index, and the glutathione S-transferase activity was assayed towards ETHA (class Pi) and CDNB (several classes). Mutagenicity was evaluated by the Ames test using Salmonella typhimurium strains TA97, TA98, TA100, and TA102.

Results

The anti-inflammatory effects of EFBt were higher than those of AEBt or AFBt. Mice treatment (3-300 μg/kg) with AFBt reduced the paw edema (3 h) at lower levels (29.2-37.3%; P < 0.01), than those observed for AEBt (44.7-54.2%; P < 0.001), EFBt (49.3-58.2%; P < 0.001) or indomethacin (64.6%, P < 0.001, 10 mg/kg). The growth of kidney cells (HEK) was inhibited by AEBt (IC₅₀ 182.6 μg/ml), EFBt (IC₅₀ 78.1 μg/ml) and AFBt (IC₅₀ 86.2 μg/ml), with lower effects on HTC hepatic cell (IC₅₀ 308.8 μg/ml, 396.5 μg/ml and 167.9 μg/ml, respectively). As evaluated by MTT test, AFBt exhibited cytotoxicity for HEK cells (IC₅₀ 372.5 μg/ml), but none for HTC ones; by the way, AFBt stimulated spleen cells (EC₅₀ 2.2 μg/ml) while cyclosporine, a cytotoxic reference drug inhibited them with IC₅₀ of 0.42 μg/ml; the IC₅₀ for doxorubicin for HEK and HTC cells was 0.28 μg/ml and 14.4 μg/ml, respectively, at 96 h. No mutagenic potential was observed. Mice treatment with AEBt or AFBt at 42 mg/kg for 15 days altered the kidney relative weight, but not at 4.2 mg/kg. Baccharis trimera did not change liver, spleen or popliteal lymph node relative weight. DNA damage index of kidney cells was observed on mice treated with AEBt/AFBt, but not on animals treated with EFBt, while DNA lesions were detected on liver cells only after AFBt treatment. The general activities of hepatic GST and Pi GST were reduced by EFBt and AFBt treatment, respectively.

Conclusions

Baccharis trimera did not show mutagenicity, inhibited the GST activity, a hepatic detoxification enzyme, and induced in vivo (genotoxicity) and in vitro toxicological effects to kidney cells.     

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.     

Antipyretic, analgesic, and anti-inflammatory activities of the seaweed Sargassum fulvellum and Sargassum thunbergii in mice

Dichloromethane, ethanol, and boiling water extracts of the brown seaweeds Sargassum fulvellum and Sargassum thunbergii were examined for antipyretic, analgesic, and anti-inflammatory activities in mice. The activities were evaluated against yeast-induced pyrexia, tail-flick test, and phorbol myristate acetate-induced inflammation (edema, erythema, and blood flow). The dichloromethane extract (0.4 mg/ear) of Sargassum fulvellum inhibited an inflammatory symptom of mouse ear edema by 79.1%. The ethanol extract (0.4 mg/ear) of Sargassum thunbergii also inhibited edema by 72.1%. No acute toxicity was observed after p.o. administration of each extract (5 g/kg bw). These findings are consistent with various claims that these seaweeds can be used as remedies for inflammation-related symptoms.     

Anti-inflammatory activity of phylligenin, a lignan from the fruits of Forsythia koreana, and its cellular mechanism of action

AIM OF THE STUDY: The fruits of Forsythia koreana have long been used in Chinese medicine to treat inflammatory disorders. However, the pharmacological data is not sufficient to clearly establish a scientific rationale for the anti-inflammatory medicinal use of this plant material, and the search for its active principles has been limited so far. MATERIALS AND METHODS: Phylligenin (lignan) was isolated from the fruits of Forsythia koreana and its anti-inflammatory activity was examined. RESULTS AND CONCLUSION: Phylligenin (1-100 microM) and the methanol extract of Forsythia koreana fruits inhibited cyclooxygenase-2 (COX-2)-mediated prostaglandin E(2) and inducible nitric oxide synthase (iNOS)-mediated nitric oxide (NO) synthesis from lipopolysaccharide-treated RAW 264.7 cells. In the mechanism study, phylligenin inhibited iNOS expression and nuclear factor-kappaB (NF-kappaB) activation but had no effect on COX-2 expression. Moreover, phylligenin significantly inhibited mouse carrageenan-induced paw edema by intraperitoneal administration (22.1-34.7% inhibition at 12.5-100 mg/kg). These pharmacological properties indicate that phylligenin possesses significant anti-inflammatory activity in vitro and in vivo, and may provide the scientific rationale for anti-inflammatory use of the fruits of Forsythia koreana.     

Studies on the analgesic,anti-inflammatory and antipyretic effects of Parquetina nigrescens leaf extract

Ethnopharmacological relevance

Parquetina nigrescens is a shrub that is commonly used in different parts of West Africa for the treatment of several ailments which includes pain, fever and inflammatory conditions.

Aim of the study

The present study was designed to investigate the analgesic, anti-inflammatory and antipyretic effects of the aqueous extract of Parquetina nigrescens leaves in rats.

Materials and methods

Five groups were used for each study, groups 1 and 5 served as control (saline) and reference (indomethacine) respectively, while groups 2–4 received the extract (50–200 mg/kg) orally. Formalin paw licking and hot plate latency tests were used for analgesic studies. Carrageenan oedema, cotton pellet granuloma and formaldehyde arthritis models were used to quantify the anti-inflammatory activities while the brewer’s yeast was used for inducing pyrexia.

Results

The results of the analgesic study show that the extract produced significant (p < 0.05) analgesia in the hot plate and in the formalin tests. In the anti-inflammatory study, Parquetina nigrescens produced significant (p < 0.05) inhibition of the various types of inflammation. The extract also inhibited the pyrexia induced by brewer’s yeast.

Conclusion

The result justifies the traditional uses of Parquetina nigrescens for the treatment of fever, inflammatory and painful conditions.     

From popular use to pharmacological validation: A study of the anti-inflammatory,anti-nociceptive and healing effects of Chenopodium ambrosioides extract

Ethno-pharmacological relevance

Chenopodium ambrosioides (Amarantaceae) is an annual or perennial plant popularly known as ‘erva de Santa Maria’, ‘mastruço’ and ‘erva-do-formigueiro’. This herb is used in folk medicine in the form of teas, poultices and infusions for inflammatory problems, contusions and lung infections, and as an anthelmintic and anti-fungal.

Aim of the study

The aim of the present study was to further the understanding of the anti-nociceptive, anti-inflammatory and wound healing effects of ethanol extract (EE) obtained from the leaves and stems of Chenopodium ambrosioides in animal models of acute pain, inflammation and wound healing, thus supporting its medicinal use for the treatment of pain and inflammatory conditions

Materials and methods

The anti-nociceptive activity of EE (150–500 mg/kg) was evaluated using the nociception induced by formalin (2.5%), prostaglandin-E₂ (PGE2; 3 nmol/paw), capsaicin (CAP, 1.6 μg/paw) and bradykinin (BK, 10 nmol/paw). The anti-inflammatory activity of EE (150–500 mg/kg) was evaluated in carrageenan- (Cg, 300 μg/paw), PGE₂- (3 nmol/paw), substance P- (SP, 20 nmol/paw) and BK- (3 nmol/paw) induced paw oedema. The topical anti-inflammatory activity of EE (1%, 3% and 5%) was evaluated in arachidonic acid- (AA, 2 mg/ear), oil croton- (1 μg/ear) and CAP- (250 μg/ear) induced ear oedema. The effect of this extract in the inhibition of the influx of neutrophil, myeloperoxidase (MPO) and adenosine-deaminase (ADA) activities and nitric oxide (NO) and TNF-á levels was also determined using the mouse of pleurisy induced by Cg. The excision wound model in rats was used to evaluate the wound healing efficacy of EE (1%, 3% and 5%). To exclude the possible non-specific muscle relaxant or sedative effects of EE, mice motor performance was also evaluated with the rota-rod test.

Results

EE (5% per ear) was effective in reducing ear oedema induced by croton oil by 78.09%, CAP by 70.85% and AA by 77.02%. EE (500 mg/kg; p.o.) also significantly inhibited paw oedema induced by Cg by 40%, PGE₂ by 51%, SP by 56% and BK by 57%. EE (500 mg/kg; p.o.) inhibited the cell influx of leucocytes by 78% and neutrophils by 53%, MPO activity by 62.22% and ADA activity by 23.07%, as well as NO by 77.77% and TNF-á levels by 50% in the fluid leakage due to the carrageenan-induced pleurisy. EE also inhibited the formalin-induced nociceptive in both phases of pain (neurogenic and inflammatory) at a dose of 500 mg/kg, resulting in inhibitions of 77.39% and 95.60%, respectively. EE (500 mg/kg; p.o.) was also effective in inhibiting the nociception induced by PGE₂ (68%), CAP (53%) and BK (32%). Topical application of EE (5%) on excision wounds caused a significant reduction in wound area when compared with the untreated controls. Finally, treatment with EE (150–500 mg/kg) did not show any significant alterations in motor performance or body temperature compared with the control group.

Conclusions

The results, including the inhibition of mediators (BK, NO, SP, PGE₂ and TNF-á) and enzyme (MPO and ADA) activity, validate the use of the plant under study for therapeutic treatment of anti-inflammatory, painful and wound healing processes.     

Involvement of heme oxygenase-1 in the anti-inflammatory activity of Chrysanthemum boreale Makino extracts on the expression of inducible nitric oxide synthase in RAW264.7 macrophages

Aim of the study

This study is to elucidate the involvement of anti-inflammatory heme oxygenase-1 (HO-1) in the inhibitory activity of a Chrysanthemum boreale Makino (CB) extract on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages.

Materials and methods

Cell viability and NO assay were performed. In addition, iNOS expression was detected by Western blotting and real-time PCR. HO-1 expression was also evaluated by Western blotting, and blocking HO-1 activity on NO production was performed.

Results

The CB extract at the highest concentration (100 μg/ml) significantly inhibited NO production by approximately 90% and suppressed iNOS protein expression by approximately 84.8% compared to LPS-stimulated cells. Furthermore, the CB extract (100 μg/ml) inhibited iNOS mRNA expression in a concentration-dependant manner and suppressed iNOS mRNA expression by 94.8%. The CB extract induced the expression of HO-1 in a dose-dependent manner, and blocking HO-1 activity abolished the inhibitory effects of the CB extract. Moreover, the addition of carbon monoxide such as tricarbonyl dichlororuthenium (II) dimmer (RuCO), a byproduct derived from heme degradation, mimicked the inhibitory action of low concentrations of CB extract.

Conclusion

These results suggest that a CB extract has potent anti-inflammatory activity in RAW264.7 macrophages involving the induction of HO-1.     

Elucidation of the transport mechanism of baicalin and the influence of a Radix Angelicae Dahuricae extract on the absorption of baicalin in a Caco-2 cell monolayer model

Ethnopharmacological relevance

Angelicae Dahurica (Hoffm.) Benth. & Hook. f. ex Franch. & Sav combined with Radix Scutellariae baicalensis Georgi has been widely used in traditional Chinese medicine (TCM) as an antipyretic analgesic and anti-inflammatory drug. Modern pharmacological studies have demonstrated that the compatible application of these two drugs is an effective treatment for hepatitis. A previous study indicated that a Radix Angelicae Dahuricae extract enhanced the intestinal absorption of the baicalin found in Radix Scutellariae; however, the underlying compatibility mechanism of these two herbs remains unknown. In this study, we further examined the effect of a Radix Angelicae Dahuricae extract on the absorption and transport properties of baicalin in a Caco-2 cell model to determine the compatibility mechanism of these two herbs.

Aim of the study

The aim of this work was to study the transport properties of baicalin in Radix Scutellariae across cell membranes and the effects of a Radix Angelicae Dahuricae extract on baicalin absorption using the well-characterized, human-based intestinal Caco-2 cell model.

Materials and methods

We assessed the absorption, bidirectional transport and toxicity of baicalin using a range of parameters, including drug concentration, pH, a P-glycoprotein (P-gp) inhibitor (Verapamil), an MRP inhibitor (MK-571) and EDTA-Na₂ (tight junction modulator). Next, we studied the influence of a Radix Angelicae Dahuricae extract on the transport of baicalin under the same conditions. Drug concentration was measured by HPLC, and the apparent permeability coefficient (Papp) and apparent permeability ratio (PDR) were subsequently calculated.

Results

The results showed that baicalin is non-toxic within a concentration range of 800 µg/mL to 4800 µg/mL. The transport of baicalin showed time and concentration dependence. The absorption of baicalin was optimal at pH 7.4 in 37 °C; however, the absorption decreased at 4 °C. The Papp of baicalin transport through the Caco-2 cell monolayer model was altered when specific inhibitors of P-gp or MRP were added to the cells. However, there was no significant difference in the PDR value. The Papp of baicalin improved when it was combined with the Radix Angelicae Dahuricae extract. The influence of EDTA-Na₂ on the transport of baicalin showed that the permeability of baicalin significantly increased. The result further indicated that the mechanism of baicalin intestinal absorption in the Caco-2 cell monolayer involves passive transcellular diffusion.

Conclusions

Passive diffusion is the main mode of intestinal absorption of bacalin and it involved in the efflux of proteins. The enhanced intestinal absorption of baicalin by Radix Angelicae Dahuricae can be due to opening of the tight junctions between cells and inhibition of MRP efflux protein expression or function.     

Preliminary study of antidiabetic activity of the methanolic leaf extract of Axonopus compressus (P. Beauv) in alloxan-induced diabetic rats

Ethnopharmacological relevance

Axonopus Compressus is commonly used by the people of Southern Nigeria to treat different ailment such as common cold and diabetes. This study therefore, evaluated the anti-diabetic effect of the methanolic leaf extract of the plant.

Materials and methods

Diabetes was induced in the rats by intraperitoneal (i.p.) injection of alloxan monohydrate at the dose of 180 mg/kg. Three test doses of the extract (250, 500 and 1000 mg/kg) administered per os through gastric gavage to the rats were used in the study. The activity was compared to a standard reference drug (glibenclamide, 2 mg/kg) and a negative control. Blood from the tail snip was used to measure the effects of the extract and drug at 0, 1, 3 and 6 h using autoanalyzer (AccuCheck Active^®) glucose kit.

Results

Methanolic leaf extract of Axonopus compressus at all the doses (250, 500 and 1000 mg/kg) used caused a respective time dependent and significant (p < 0.0001) reduction (by 31.5%, 19.8% and 24.5%) of the blood glucose levels in the diabetic rats when compared to the negative control group at the 6th hour. However, the reference drug (glibenclamide, 2 mg/kg) decreased the blood glucose levels by 69.9% and the tween 20 solution (negative control) increased the blood glucose level by 15.2% at the 6th hour. Moreso, the extract at the different test doses caused various degrees of reduction of the blood glucose levels of the test rats at 1st, 3rd and 6th hours when compared to the negative control rats.

Conclusion

The findings suggest that Axonopus compressus may possess antidiabetic property.     

天王补心丸中生地黄、玄参、天冬和麦冬水提液对大鼠肝CYP450酶含量及其亚型CYP3A, CYP2E1和CYP1A2活性的影响

目的:研究天王补心丸中滋阴类药味生地黄、玄参、天冬和麦冬水提物对大鼠肝肝药酶(CYP450)含量及对亚型CYP3A,CYP2E1,CYP1A2活性的影响,探讨CYP450在天王补心丸代谢中的作用.方法:大鼠灌胃给予生地黄、玄参、天冬和麦冬水提物7d后取肝脏称重并制备肝微粒体,紫外分光光度法测定肝微粒体细胞色素b5(Cytb5),P450的含量及CYP3A的活性,高效液相法测定CYP2E1和CYP1A2的活性.结果:与空白对照组比较,各给药组大鼠肝指数无显著变化;生地黄组CYP450含量极显著减少(P＜0.01),CYP3A活性极显著增加(P＜0.01),CYP1A2活性显著增加(P＜0.05);玄参组CYP450含量减少(P＜0.05),CYP3A和CYP1A2活性均极显著增加(P＜0.01);天冬组Cytb5含量增加(P＜0.05),CYP2E1活性增加(P＜0.05),CYP1A2活性极显著增加(P＜0.01);麦冬组cytb5,CYP450含量无统计学差异,CYP3A活性增加(P＜0.05),CYP2E1活性增加(P＜0.05),CYP1A2活性极显著增加(P＜0.01).结论:天王补心丸中生地黄、玄参降低大鼠肝脏CYP450酶含量并均诱导CYP3A和CYP1A2活性,天冬诱导CYP2E1和CYP1A2活性,麦冬诱导CYP3A,CYP2E1和CYP1A2活性.由于抑制CYP450活性,减少对其他药代谢,滋阴药组在天王补心丸全方配伍中可增强其他各功能组比如补血养血、宁心安神类药味的作用,为探讨天王补心丸的配伍机制提供了理论基础.     

In vitro and in vivo anti-inflammatory effects of taheebo, a water extract from the inner bark of Tabebuia avellanedae

AIM OF STUDY: Tabebuia spp. (Bignoniaceae) are native to tropical rain forests throughout Central and South America and have long been used as a folk medicine to treat bacterial infection, blood coagulation, cancer and inflammatory diseases. In this study, we aimed to demonstrate the ethnopharmacological activity of Tabebuia avellanedae in various in vitro and in vivo inflammatory conditions. MATERIALS AND METHODS: To do this, LPS-stimulated macrophages and arachidonic acid or croton oil-induced mouse ear edema models were employed. RESULTS: The water extract (taheebo) of Tabebuia avellanedae significantly suppressed the production of prostaglandin (PG) E(2) and nitric oxide (NO), and blocked the mRNA expression of their catalyzing enzymes (cyclooxygenase [COX)-II] and inducible NO synthase [iNOS], respectively), in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The blockade of inflammatory mediators by taheebo seemed to be the result of the interruption of extracellular signal-related kinase (ERK) activation, according to immunoblotting analysis and the NO assay, where LPS strongly induced the phosphorylation (a hallmark of activation) of ERK, and U0126, a selective ERK inhibitor, was found to strongly inhibit PGE(2) production. Similarly, oral administration of taheebo (100mg/kg) for 1 week completely diminished mouse ear edema induced by arachidonic acid, an activator of COX-II, but not croton oil, an activator of lipoxygenase. CONCLUSIONS: These data suggest that the ethnopharmacological action of taheebo may be due to its negative modulation of macrophage-mediated inflammatory responses by suppressing PGE(2) production. Thus, this water extract may be developed as a new therapeutic remedy for various inflammatory diseases such as arthritis and atherosclerosis.     

Diuretic activity and kidney medulla AQP1, AQP2, AQP3, V2R expression of the aqueous extract of sclerotia of Polyporus umbellatus FRIES in normal rats

Aim of the study

Zhuling, sclerotia of Polyporus umbellatus FRIES, a Traditional Chinese Medicine, has long been used as a diuretic. The aim of the present study was to evaluate the diuretic effect on the urinary electrolyte concentration (Na⁺, K⁺, and Cl⁻) and regulation of the relative mRNA expression of aquaporin-1 (AQP1), aquaporin-2 (AQP2), aquaporin-3 (AQP3) and vasopressin V₂ receptor (V₂R) post-oral administration of sclerotia of Polyporus umbellata aqueous extract in normal rats.

Materials and methods

Aqueous extract of sclerotia of Polyporus umbellatus (50 mg/kg, 250 mg/kg, 500 mg/kg) or the reference drug, furosemide (10 mg/kg) were administrated orally to male SD rats and their urine output was quantified and collected 24 h and 8 days after the treatment. The kidney medulla AQP1, AQP2, AQP3 and V₂R mRNA relative expressions were measured with RT-PCR.

Results

After single dose of the exact of sclerotia of Polyporus umbellata, urine output was found to be significantly increased, which began at 4 h, and at 24 h after the treatment, the sclerotia of Polyporus umbellatus extract and furosemide treatment produced the similar total volume of urine excreted. The extract increases urinary levels of Na⁺, K⁺, and Cl⁻, to about the same extent, while furosemide increased urinary levels of Na⁺ and Cl⁻. After the 8-day doses, all two substances induced significant diuresis, natriuresis and chloriuresis. These two substances do not regulate the AQP1 and AQP3 mRNA level in normal rat kidney medulla. The AQP2 mRNA level of sclerotia of Polyporus umbellata extract was down-regulated significantly, the V₂R mRNA level of sclerotia of Polyporus umbellata extract 50 mg/kg dose group and 250 mg/kg dose group were down-regulated significantly too. Interestingly, the low-dose group had higher effect on regulation of AQP2 and V₂R mRNA level.

Conclusion

Aqueous extract of sclerotia of Polyporus umbellatus has conspicuous diuretic effect confirming its ethnopharmacological use. From the pattern of excretion of water, sodium, potassium, chlorine, AQP2 and V2R mRNA level, it may be logically concluded that it has effect from down-regulating AQP2, and down-regulate AQP2 by down-regulating V₂R.     

Antisecretory activity from the flowers of Chiranthodendron pentadactylon and its flavonoids on intestinal fluid accumulation induced by Vibrio cholerae toxin in rats

Ethnopharmacological relevance

The flowers of Chiranthodendron pentadactylon Larreat. (Sterculiaceae) has been traditionally used as folk medicine in Mexico for the treatment of gastrointestinal disorders such as diarrhea and dysentery.

Aim of the study

This study aimed to assess the antisecretory activity which supports the therapeutic use of Chiranthodendron pentadactylon and its flavonoids to treat diarrhea.

Materials and methods

The methanol extract of Chiranthodendron pentadactylon, subsequent fractions, and flavonoids were evaluated on cholera toxin-induced intestinal secretion in rat jejunal loops model.

Results

Three antisecretory flavonoids were isolated by bioassay-guided purification, namely, isoquercitrin 3, (+)-catechin 4 and (−)-epicatechin 5. Among them, epicatechin exhibited the most potent antisecretory activity with ID₅₀ of 8.3 μM/kg. Its potency was close that of to loperamide (ID₅₀ 6.1 μM/kg), drug used as control. Isoquercitrin (ID₅₀ 19.2 μM/kg) and catechin (ID₅₀ 51.7 μM/kg) showed moderate and weak activity, respectively.

Conclusion

The results of the present study lend some support to the anecdotal report for the traditional use of the flowers of Chiranthodendron pentadactylon in the control of dysentery.     

Anti-ulcer activity of Cyrtocarpa procera analogous to that of Amphipterygium adstringens, both assayed on the experimental gastric injury in rats

Ethnopharmacological relevance

The bark of Amphipterygium adstringens (Aa) is commonly mixed or adulterated with the bark of Cyrtocarpa procera (Cp) and sold in Mexican markets. Aa is a well known species in Mexico used as decoction to relieve ulcers. Scientific reports reinforcing the anti-ulcer activity of Aa have been previously described, but those describing the anti-ulcer properties of Cp as a substitute for Aa in folk medicine are scarce.

Aim of the study

To investigate anatomical and phytochemical differences between these species, as well as to assess the anti-ulcer effect of Cp extracts in comparison to the Aa extracts.

Material and methods

Anatomical micro-technique and physical and spectroscopic data were used to analyze differences between Cp and Aa. Regard to the pharmacological activity, it was assessed by using the ethanol-induced gastric damage model in rats.

Results

Whereas the bark anatomy of Aa was characterized by vertical canals in the periderm and the rare occurrence of fibers in its phloem, a periderm without vertical canals and abundant fibers in the phloem were distinctive features of Cp. Phytochemical analysis allowed the identification of tirucallane, masticadienonic and 3α-hydroxymasticadienonic acids as major components in Aa, while β-amyrin and β-sitosterol were obtained from Cp. Gastric lesions observed in the control group decreased in the presence of 100 mg/kg of hexane, ethyl acetate and methanol extracts from the normal or regenerated bark of Cp, thus resembling the anti-ulcer effect of Aa. Nevertheless, major anti-ulcer potency was observed with the most active methanol extract from Cp obtained from normal [the effective dose fifty ED₅₀ = 45.54 mg/kg] or regenerated (ED₅₀ = 36.68 mg/kg) bark in comparison to Aa (ED₅₀ = 115.64 mg/kg).

Conclusion

Chemical and anatomical differences were found between these species, but since the anti-ulcer activity of Cp is similar to that shown by Aa our results reinforce the use of both species for the relief of gastric ulcer in folk medicine.     

Fraxinus rhynchophylla ethanol extract attenuates carbon tetrachloride-induced liver fibrosis in rats via down-regulating the expressions of uPA,MMP-2, MMP-9 and TIMP-1

Aim of the study

To investigate the effect of Fraxinus rhynchophylla ethanol extract (FR_EtOH) on liver fibrosis induced by carbon tetrachloride (CCl₄) in rats.

Materials and methods

Rat hepatic fibrosis was induced by oral administration of CCl₄. Sixty SD rats were divided randomly into 6 groups: control, CCl₄ group, silymarin group and three FR_EtOH-treated groups. Except for the rats in control group, all rats were administered orally with CCl₄ (20%, 0.2 mL/100 g body weight) twice a week for 8 weeks. Rats in FR_EtOH groups were treated daily with FR_EtOH (0.1, 0.5 and 1.0 g/kg, p.o.) throughout the whole experimental period. Liver function parameters (such as activities of serum GOT and GPT levels), activities of liver anti-oxidant enzymes (such as catalase, SOD, GPx) and expressions of uPA, tPA, MMP-2, MMP-9 and TIMP-1, -2, -3, -4 in the liver fibrosis pathway were detected.

Results

The results showed that FR_EtOH (0.1, 0.5 and 1.0 g/kg BW) significantly reduced the elevated activities of sGOT and sGPT caused by CCl₄. FR_EtOH (0.1 and 0.5 g/kg BW) and significantly increased the activities of GSH-Px. The histopathological study showed that FR_EtOH (0.1 and 0.5 g/kg BW) reduced the incidence of liver lesions, including hepatic cells cloudy swelling, lymphocytes infiltration, cytoplasm vacuolization hepatic necrosis and fibrous connective tissue proliferated induced by CCl₄ in rats. In our study it was showed that CCl₄-treated group significantly increased the protein levels of uPA, MMP-2, MMP-9 and TIMP-1. FR_EtOH (0.1 and 0.5 g/kg BW) could inhibit the protein levels of uPA, MMP-2, MMP-9 and TIMP-1. Finally, the amount of esculetin in the FR_EtOH was 33.54 mg/g extract.

Conclusions

Oral administration of FR_EtOH significantly reduces CCl₄-induced hepatic fibrosis in rats, probably by exerting a protective effect against hepatocellular fibrosis by its free radical scavenging ability. FR_EtOH down-regulated the expressions of uPA, MMP-2 and MMP-9 in CCl₄-induced liver fibrosis in rats.     

肿节风总黄酮对巨核系细胞体外扩增的作用研究

目的: 探讨肿节风总黄酮(ZJF-HT)对小鼠骨髓巨核系细胞扩增的影响。 方法: 采用液体培养和半固体集落培养,观察ZJF-HT及其含药血清对成熟巨核细胞及巨核系祖细胞集落扩增的作用。 结果: 与模型组相比,ZJF-HT≥125 μg·mL^-1能使成熟巨核细胞数和巨核系祖细胞集落数增多(P<0.05);ZJF-HT含药血清在0.39%~6.25%体积浓度范围能增加成熟巨核细胞数,在0.20%~6.25%范围内能显著增加巨核系祖细胞集落数(P<0.01)。 结论: ZJF-HT及其含药血清均能促进巨核系细胞的体外扩增。     

Transport properties of puerarin and effect of Radix Angelicae Dahuricae extract on the transport of puerarin in Caco-2 cell model

Ethnopharmacological relevance

Angelicae Dahurica (Hoffm.)Benth.& Hook.f.ex Franch.&Sav combined with Pueraria labota (Willd.)Ohwi has been widely used as herb-pairs in traditional Chinese medicine (TCM) for utilization of antipyretic analgesic and anti-inflammatory drugs, and modern pharmacological studies have shown that application compatibility of the two drugs has the effects of cardiovascular disease treatment. The previous study has proved that Radix Angelicae Dahuricae extract could enhance the intestinal absorption of puerarin in Pueraria. But the underlying compatibility mechanism of the two herbs remains unknown. In this study we tried to further evaluate the improvement of Radix Angelicae Dahuricae extract on the puerarin using the Caco-2 cell model and explore the transport properties of puerarin through the above research to discuss the possible effect mechanism of Radix Angelicae Dahuricae extract on the transport of puerarin and the underlying compatibility mechanism of the two herbs.

Aim of study

The aim of this work was to study the transport properties of puerarin in Radix Pueraria across Caco-2 cell membrane and to explore how the Radix Angelicae Dahuricae extract affected the transport of puerarin using the well-characterized, human-based intestinal Caco-2 cell model as a platform.

Materials and methods

The bidirectional transport, and the effects of time, drug concentration, pH, P-gp inhibitors (Verapamil, Cyclosporin A), MRP inhibitor (MK-571) and EDTA-Na₂ (tight junction modulator) on the absorption of puerarin were observed. Then the influence of extract of Radix Angelicae Dahuricae on the transport of puerarin was studied. Drug concentration was measured by HPLC and the apparent permeability coefficients (Papp) and apparent permeability ratio (PDR) were calculated.

Results and conclusions

The results showed that the transport (Papp) of puerarin in Caco-2 cell monolayer model had time and concentration dependence, and the transport showed saturation characteristics with the time and concentration of puerarin to a certain degree. The Papp of puerarin transported on Caco-2 cell monolayer model was significantly changed when the specified inhibitors of P-gp were added to the model and the PDR decreased from 1.74 to 0.43. The absorption of puerarin was improved when combined with Radix Angelicae Dahuricae. The intestinal absorption of puerarin is by passive diffusion as the dominating process and active transportation was mediated by P-gp and MRP transporter in Caco-2 cell monolayer model, and Radix Angelicae Dahuricae could enhance the intestinal absorption of puerarin.     

Effects of unprocessed versus vinegar-processed Schisandra chinensis on the activity and mRNA expression of CYP1A2, CYP2E1 and CYP3A4 enzymes in rats

Ethnopharmacological relevance

Schisandra chinensis (SC) is a well-known traditional Chinese herbal medicine that has been used in clinical practices for thousands of years. However, the differences between the effects of unprocessed and vinegar-processed Schisandra chinensis (VSC) on cytochrome P450 (CYP450) activities are poorly understood.

Aim of the study

To evaluate the differences between processed and unprocessed SC on the metabolism of CYP1A2, CYP2E1 and CYP3A4 substrates in rats using a cocktail method based on a developed and validated HPLC method. We also investigate the influence of processing on the levels of CYP mRNA.

Materials and methods

Three probe substrates (theophylline, dapsone and chlorzoxazone) were delivered simultaneously into rats treated with single or multiple doses of processed or unprocessed SC extract. The plasma concentrations of the three probes were profiled by HPLC, and their corresponding pharmacokinetic parameters were calculated. Real-time RT-PCR was performed to determine the effects of processed and unprocessed SC on the mRNA expression of CYP1A2, CYP2E1 and CYP3A4 in the liver.

Results

Treatment with single or multiple doses of either extract of SC induced CYP3A4 enzyme activity and inhibited CYP1A2 enzyme activity in rats. Furthermore, the inhibitory effect of SC was more potent after vinegar processing than without vinegar processing. CYP2E1 enzyme activity was induced after treatment with a single dose but was inhibited after multiple doses. The mRNA expression results were in accordance with the pharmacokinetic results.

Conclusions

These results provide useful scientific data for the safe clinical application of either extract of SC in combination with other drugs, which should lack the side effects induced by other herb–drug interactions.