The effects of Buyang Huanwu Decoction on hemorheological disorders and energy metabolism in rats with coronary heart disease

Ethnopharmacological relevance

Buyang Huanwu Decoction (BYHWD), a traditional Chinese medicine (TCM) formula, has been recognized as a clinical treatment for coronary heart disease (CHD) with qi deficiency and blood stasis syndrome. The effects of BYHWD on hemorheological disorders and energy metabolism in CHD with qi deficiency and blood stasis syndrome are still unclear.

Aim of the study

To investigate whether the ameliorative effects of BYHWD on CHD rats with qi deficiency and blood stasis syndrome are associated with the regulation of hemorheological disorders and energy metabolism.

Materials and methods

The rats were lavaged with 25.68, 12.84 and 6.42 g/kg BYHWD (g weight of mixed crude drugs/kg body weight), respectively, once a day for 21 days. The body weight, exhaustive swimming time and tongue characters were observed and recorded. The whole blood viscosity and plasma viscosity were determined by hematology analyzer. The level of fibrinogen (Fbg) in plasma was determined by using Fbg assay kit. The platelet aggregation induced by adenosine diphosphatase was measured by semi-automatic whole blood platelet analyzer. The level of blood glucose (BG) was determined by LifeScan. The activity of Na⁺-K⁺-ATPase in heart tissues was detected by spectrophotometer.

Results

BYHWD improved the exterior signs of qi deficiency and blood stasis syndrome in rats with CHD, including the body weight, exhaustive swimming time and tongue quality. The whole blood viscosity in rats treated with 25.68 g/kg BYHWD decreased at the shear rate of 10 s⁻¹ (P < 0.05) and the plasma viscosity decreased in rats treated with 25.68 and 12.84 g/kg BYHWD (P < 0.05). The plasma Fbg level and the platelet aggregation decreased in rats treated with 25.68 g/kg BYHWD (P < 0.01). The results also revealed that the BG level decreased and the Na⁺-K⁺-ATPase activity in heart tissues increased in rats treated with 25.68 and 12.84 g/kg BYHWD (P < 0.01).

Conclusion

The results suggest that the ameliorative effects of BYHWD on CHD rats with qi deficiency and blood stasis syndrome are mediated by the improvement of hemorheological disorders and energy metabolism.     

Apocynum venetum leaf aqueous extract inhibits voltage-gated sodium channels of mouse neuroblastoma N2A cells

Ethnopharmacological relevance

Apocynum venetum Linn. (Apocynaceae family), also called Luobuma, is a shrub which grows widely in the Xinjiang Autonomous Region of China. Its leaves are used in herbal tea for the treatment of hypertension, anxiety and depression. Animal studies have also shown that Apocynum venetum leaf extract (AVLE) also exerts anti-depressant and anti-anxiety activities. The effects of AVLE on neuronal tissues in vitro are not fully understood.

Materials and methods

Using the whole-cell voltage-clamp method, we studied the effects of AVLE on ion channels in cultured mouse neuroblastoma N2A cells.

Results

AVLE inhibited voltage-gated inward Na⁺ current in a reversible and concentration-dependent manner (half-inhibitory concentration was 18 μg/ml and maximum inhibition at 100 μg/ml). AVLE specifically promoted steady-state inactivation of Na⁺ channels but did not affect voltage-dependence of activation. The inhibitory effect was not use-dependent and was not affected by 300 μM L-NAME, suggesting that NO was not involved in the action of AVLE in neuronal cells. AVLE also had a mild inhibitory effect on voltage-gated K⁺ channels, but did not affect ATP-sensitive K⁺ channels.

Conclusions

Since voltage-gated Na⁺ and K⁺ channels are associated with neuronal excitability and therefore affect neurotransmission, the modulation of neuronal ion channels by AVLE may exert neuropharmacological effects. In particular, the inhibition of voltage-gated Na⁺ currents by AVLE may in part account for the psychopharmacological effects of this herbal remedy.     

An aqueous birch leaf extract of Betula pendula inhibits the growth and cell division of inflammatory lymphocytes

Ethnopharmacological relevance

Leaf extracts of Betula pendula have been traditionally used for the treatment of patients with rheumatoid arthritis (RA) or osteoarthritis.

Aim of the study

We investigated the anti-proliferative capacity of an aqueous leaf extract of Betula pendula (BPE) on human primary lymphocytes in vitro, because activated lymphocytes play a major role in the initiation and maintenance of RA.

Materials and methods

Lymphocyte proliferation and cell division was measured by the activity of mitochondrial dehydrogenases and by using the membrane-permeable dye carboxyfluorescein diacetate succinimidyl ester (CFSE), respectively. Apoptosis was analyzed by surface staining of phosphatidylserine and intracellular activation of effector caspases 3 and 7 in comparison to the drug methotrexate using flow cytometric and photometrical analysis. In addition, the impact of the extract on cell cycle distribution was investigated by propidium iodide staining of DNA. For the bioassays BPE concentrations of 10-160 μg/mL were investigated. A phytochemical analysis, using LC-MS and HPLC, was conducted to identify the polyphenolic constituents of the birch leaf extract.

Results

Leaf extracts of Betula pendula inhibited the growth and cell division (CD8⁺: 40 μg/mL: 45%; 80 μg/mL: 60%; 160 μg/mL: 87%) (CD4⁺: 40 μg/mL: 33%; 80 μg/mL: 54%; 160 μg/mL: 79%) of activated, but not of resting T lymphocytes in a significant dose-dependent manner. The inhibition of lymphocyte proliferation due to apoptosis induction (compared to untreated control: 40 μg/mL: 163%; 80 μg/mL: 240%; 160 μg/mL: 348%) and cell cycle arrest was comparable to that of methotrexate. LC-MS analyses showed that the extract contains different quercetin-glycosides.

Conclusion

Our results give a rational basis for the use of Betula pendula leaf extract for the treatment of immune disorders, like rheumatoid arthritis, by diminishing proliferating inflammatory lymphocytes.     

Protective effects of Polygonatum sibiricum polysaccharide on ovariectomy-induced bone loss in rats

Ethnopharmacological relevance

To assess the ability of traditional Chinese medicine Polygonatum sibiricum polysaccharide to prevent bone loss in the ovariectomized rat.

Materials and methods

PSP was administered intragastrically to the rats. After 35 days, the total body bone mineral density (BMD) was assessed in all of the rats. All sections were processed for immunohistochemistry and hematoxylin-eosin staining (H.E.).

Results

BMD was lower in the ovariectomized group (OVX, 0.163 g/cm²), the group that received a moderate dose of PSP on OVX animals (OVX + MP, 0.163 g/cm²) and the group that received a low dose of PSP on OVX animals (OVX + LP, 0.162 g/cm²) than in the sham-operated group (SHAM, 0.180 g/cm²), the OVX + E₂ group (OVX + E₂, 0.176 g/cm²) and the group that received a high dose of PSP on OVX animals (OVX + HP, 0.174 g/cm²) (P < 0.05). Clear arrangements of bone trabeculae were observed in the OVX + E₂ and OVX + HP. The expression of bone morphogenetic proteins (BMP) and basic fibroblast growth factor (bFGF) in the OVX, OVX + MP and OVX + LP was down regulated compared to the SHAM, OVX + E₂ and OVX + HP (P < 0.05). The rats in the OVX + E₂ and OVX + HP had lower levels of bone Gla protein (BGP), bone alkaline phosphatase (BALP), tartrate-resistant acid phosphatase (TRAP) and tumor necrosis factor α(TNF-α) expression than the rats in the OVX, OVX + MP and OVX + LP (P < 0.05).

Conclusion

This experiment demonstrates that the administration of PSP to ovariectomized rats reverses bone loss and prevents osteoporosis.     

The effect of Nigella sativa alone, and in combination with dexamethasone, on tracheal muscle responsiveness and lung inflammation in sulfur mustard exposed guinea pigs

Ethnomedical relevance

The anti-inflammatory activity of both systemic and local administrations of essential oil from Nigella sativa L. has been shown.

Aim of the study

Therefore, the effect of Nigella sativa on tracheal responsiveness (TR) and lung inflammation of sulfur mustard (SM) exposed guinea pigs was examined.

Materials and methods

Guinea pigs were exposed to diluent solution (control group), inhaled SM (SME group), SME treated with Nigella sativa (SME + N), SME treated with dexamethasone (SME + D) and SME treated with both drugs (SME + N + D), (n = 7 for each group). TR to methacholine, total white blood cell (WBC) and differential WBC count of lung lavage, and serum cytokines were measured 14 days post-exposure.

Results

The values of TR, eosinophil, monocyte, lymphocyte, interleukine-4 (IL-4) and interferon gamma (IFN-γ) of SME group were significantly higher than those of controls (p < 0.05 to p < 0.001). The TR in SME + N, SME + D and SME + N + D was significantly lower compared to that of SME group (p < 0.01 for all cases). The percentage of eosinophil in SME + D, and the percentage of monocyte in SME + N + D (p < 0.05 to p < 0.01) were significantly lower than those in SME group. The neutrophil number was decreased in SME + N and SME + N + D groups compared to SME animals (p < 0.05 to p < 0.01). IL-4 levels in serum of SME + N (p < 0.01), SME + D (p < 0.05), SME + N + D (p < 0.01) and IFN-γ in SME + N (p < 0.05) were greater than those in SME animals.

Conclusions

These results showed a preventive effect of Nigella sativa on TR and lung inflammation of SM exposed guinea pigs.     

Mechanisms of the relaxant effect of a Danshen and Gegen formulation on rat isolated cerebral basilar artery

Aim of the study

Danshen (root of Salvia miltiorrhiza) and Gegen (root of Pueraria lobata) are two herbs used in traditional Chinese medicine, most commonly for their putative cardioprotective and anti-atherosclerotic effects. In this study, the actions of a Danshen and Gegen formulation (DG; ratio 7:3) were investigated on rat-isolated cerebral basilar artery.

Materials and methods

Rat basilar artery rings were precontracted with 100 nM U46619. Involvement of endothelium-dependent mechanisms was investigated by mechanical removal of the endothelium; K⁺ channels were investigated by pretreatment of the artery rings with various K⁺ channel inhibitors, and Ca²⁺ channels were investigated in artery rings incubated with Ca²⁺-free buffer and primed with 100 nM U46619 for 5 min prior to adding CaCl₂ to elicit contraction.

Results

DG produced concentration-dependent relaxation of the artery rings with an IC₅₀ of 895 ± 121 μg/ml. Mechanical removal of the endothelium or pretreatment with the BK_Ca channel inhibitor iberiotoxin (100 nM), the K_V channel inhibitor 4-aminopyridine (1 mM), or the K_IR channel inhibitor barium chloride (100 μM), all had no effect on the DG-induced response (P > 0.05 for all). However, pretreatment with the K_ATP channel inhibitor glibenclamide (1 μM), the non-selective K⁺ channel inhibitor tetraethylammonium (TEA, 100 mM), or a combination of all the K⁺ channel inhibitors (iberiotoxin + 4-aminopyrindine + barium chloride + glibenclamide + TEA) produced significant inhibition on the DG-induced response (P < 0.01 for all); its maximum vasorelaxant effect (Imax) was reduced by 37, 24, and 30%, respectively. Preincubation of the artery rings with DG for 10 min produced concentration-dependent (1, 3 and 7 mg/ml) and total inhibition on the CaCl₂-induced vasoconstriction.

Conclusions

These findings suggest the vasorelaxant effect of DG on rat basilar artery is independent of endothelium-derived mediators, whereas, inhibition of Ca²⁺ influx in the vascular smooth muscle cells is important, and a minor component is mediated by the opening of K_ATP channels. DG could be a useful cerebroprotective agent in some patients with occlusive cerebrovascular disease.     

Neuroprotective effect of Momordica charantia in global cerebral ischemia and reperfusion induced neuronal damage in diabetic mice

Ethnopharmacological relevance

Momordica charantia L. (Cucurbitaceae) fruits have been used traditionally for centuries, especially for treating diabetes and associated complications.

Aim of the study

The present study was performed to evaluate neuroprotective effect of lyophilized M. charantia fruit juice against global cerebral ischemia and reperfusion induced neuronal injury in diabetic mice.

Materials and methods

Global cerebral ischemia induced by occluding both common carotid arteries for 10 min followed by 24 h reperfusion was used to induce neuronal injury. Ischemia-reperfusion induced neuronal injury was evaluated in terms of cerebral infarct size, generation of free radicals measured as thiobarbaturic acid reactive substances (TBARS), and neurological functions measured as short term memory and motor activity.

Results

The cerebral oxidative stress and damage, and neurological deficits were dose dependently attenuated by pre-treatment with the lyophilized M. charantia juice (200-800 mg/kg, p.o., o.d.). Moreover, M. charantia also exhibited dose dependent antihyperglycemic activity in diabetic mice.

Conclusions

These results suggest that M. charantia has potent neuroprotective activity against global cerebral ischemia-reperfusion induced neuronal injury and consequent neurological deficits in diabetic mice.     

Xiao-Ban-Xia-Tang inhibits cisplatin-induced pica by down regulating obestatin in rats

Ethnopharmacological relevance

Xiao-Ban-Xia-Tang (XBXT), a traditional Chinese herbal medicine, has been used in China for more than 2000 years, and proved to be effective in various cases of vomiting in the clinic.

Objective

To investigate the inhibitive effect of XBXT on cisplatin-induced pica behaviour and its effective mechanism on obestatin, CCK and CGRP in the pica model of rat.

Materials and Methods

The inhibitive effect of XBXT was investigated in the pica model of rats induced by cisplatin (3 mg kg⁻¹, i.p.) in 72 h observation, the expression of obestatin in the area postrema and ileum was measured by immunohistochemistry and PCR, and the levels of CCK and CGRP in blood were measured by Elisa.

Results

The weight of kaolin eaten in rats induced by cisplatin was significantly reduced by pretreatment with XBXT in a dose-dependent manner during the 0-24 h and 24-72 h periods (P < 0.05). XBXT exhibited effective dose-dependent (P < 0.05) inhibition on the increase of expression levels of obestatin in both the ileum and area postrema, and markedly suppressed the increase levels of CCK and CGRP in blood induced by cisplatin in a dose-dependent manner (P < 0.05).

Conclusions

XBXT has good activity against cisplatin-induced eating kaolin in rats possibly by inhibiting central or peripheral increase of obestatin, or the levels of CCK and CGRP in blood.     

Permeability of rosmarinic acid in Prunella vulgaris and ursolic acid in Salvia officinalis extracts across Caco-2 cell monolayers

Ethnopharmacological relevance

Rosmarinic acid (RA), a caffeic acid-related compound found in high concentrations in Prunella vulgaris (self-heal), and ursolic acid (UA), a pentacyclic triterpene acid concentrated in Salvia officinalis (sage), have been traditionally used to treat inflammation in the mouth, and may also be beneficial for gastrointestinal health in general.

Aim of the study

To investigate the permeabilities of RA and UA as pure compounds and in Prunella vulgaris and Salvia officinalis ethanol extracts across human intestinal epithelial Caco-2 cell monolayers.

Materials and methods

The permeabilities and phase II biotransformation of RA and UA as pure compounds and in herbal extracts were compared using Caco-2 cells with HPLC detection.

Results

The apparent permeability coefficient (P_app) for RA and RA in Prunella vulgaris extracts was 0.2 ± 0.05 × 10⁻⁶ cm/s, significantly increased to 0.9 ± 0.2 × 10⁻⁶ cm/s after β-glucuronidase/sulfatase treatment. P_app for UA and UA in Salvia officinalis extract was 2.7 ± 0.3 × 10⁻⁶ cm/s and 2.3 ± 0.5 × 10⁻⁶ cm/s before and after β-glucuronidase/sulfatase treatment, respectively. Neither compound was affected in permeability by the herbal extract matrix.

Conclusion

RA and UA in herbal extracts had similar uptake as that found using the pure compounds, which may simplify the prediction of compound efficacy, but the apparent lack of intestinal glucuronidation/sulfation of UA is likely to further enhance the bioavailability of that compound compared with RA.     

Prevention of experimental diabetes by Uncaria tomentosa extract: Th2 polarization, regulatory T cell preservation or both?

Ethnopharmacological relevance

Uncaria tomentosa (Willd.) DC (Rubiaceae) is a species native to the Amazon rainforest and surrounding tropical areas that is endowed with immunomodulatory properties and widely used around the world. In this study we investigated the immunomodulatory potential of Uncaria tomentosa (UT) aqueous-ethanol extract on the progression of immune-mediated diabetes.

Materials and methods

C57BL/6 male mice were injected with MLDS (40 mg/kg) and orally treated with UT at 10-400 mg/kg during 21 days. Control groups received MLDS alone or the respective dilution vehicle. Pancreatic mononuclear infiltrate and β-cell insulin content were analyzed by HE and immunohistochemical staining, respectively, and measured by digital morphometry. Lymphocyte immunophenotyping and cytokine production were determined by flow cytometry analysis.

Results

Treating the animals with 50-400 mg/kg of UT caused a significant reduction in the glycemic levels, as well as in the incidence of diabetes. The morphometric analysis of insulitis revealed a clear protective effect. Animals treated with UT at 400 mg/kg presented a higher number of intact islets and a significant inhibition of destructive insulitis. Furthermore, a significant protection against the loss of insulin-secreting presented β-cells was achieved, as observed by a careful immunohistochemical evaluation. The phenotypic analysis indicated that the groups treated with higher doses (100-400 mg/kg) presented CD4⁺ and CD8⁺ T-cell values similar to those observed in healthy animals. These same higher doses also increased the number of CD4⁺CD25⁺Foxp3⁺ regulatory T-cells. Moreover, the extract modulated the production of Th1 and Th2, with increased levels of IL-4 and IL-5.

Conclusions

The extract was effective to prevent the progression of immune-mediated diabetes by distinct pathways.     

Lycium barbarum polysaccharides protect mice liver from carbon tetrachloride-induced oxidative stress and necroinflammation

Ethnopharmacological relevance

Lycium barbarum has been used as a traditional Chinese medicine to nourish liver, kidneys and the eyes.

Aim of the study

We investigated the protective mechanisms of Wolfberry, Lycium barbarum polysaccharides (LBP) in carbon tetrachloride (CCl₄)-induced acute liver injury.

Materials and methods

Mice were intraperitoneally injected with a 50 μl/kg CCl₄ to induce acute hepatotoxicity (8 h) and were orally fed with LBP 2 h before the CCl₄ injection. There were six experimental groups of mice (n = 7-8 per group), namely: control mice (vehicle only; 1 mg/kg LBP or 10 mg/kg LBP), CCl₄-treated mice and CCl₄ + LBP treated mice (1 mg/kg LBP or 10 mg/kg LBP).

Results

Pre-treatment with LBP effectively reduced the hepatic necrosis and the serum ALT level induced by CCl₄ intoxication. LBP remarkably inhibited cytochrome P450 2E1 expression and restored the expression levels of antioxidant enzymes. It also decreased the level of nitric oxide metabolism and lipid peroxidation induced by CCl₄. LBP attenuated hepatic inflammation via down-regulation of proinflammatory mediators and chemokines. Furthermore, LBP promoted liver regeneration after CCl₄ treatment. The protective effects of LBP against hepatotoxicity were partly through the down-regulation of nuclear factor kappa-B activity.

Conclusion

LBP is effective in reducing necroinflammation and oxidative stress induced by a chemical toxin, thus it has a great potential use as a food supplement in the prevention of hepatic diseases.     

Antinociceptive effect of the essential oil of Zingiber zerumbet in mice: possible mechanisms

Ethnopharmacological relevance

Zingiber zerumbet (L.) Smith, a wild edible ginger species or locally known as “lempoyang”, commonly used in the Malays traditional medicine as an appetizer or to treat stomachache, toothache, muscle sprain and as a cure for swelling sores and cuts.

Aim

The present study was conducted to investigate the possible mechanism of actions underlying the systemic antinociception activity of the essential oil of Zingiber zerumbet (EOZZ) in chemical-induced nociception tests in mice.

Materials and methods

Acetic acid-induced abdominal constriction, capsaicin-, glutamate- and phorbol 12-myristate 13-acetate-induced paw licking tests in mice were employed in the study. In all experiments, EOZZ was administered systemically at the doses of 50, 100, 200 and 300 mg/kg.

Results

It was shown that EOZZ given to mice via intraperitoneal and oral routes at 50, 100, 200 and 300 mg/kg produced significant dose dependent antinociception when assessed using acetic acid-induced abdominal writing test with calculated mean ID₅₀ values of 88.84 mg/kg (80.88-97.57 mg/kg) and 118.8 mg/kg (102.5-137.8 mg/kg), respectively. Likewise, intraperitoneal administration of EOZZ at similar doses produced significant dose dependent inhibition of neurogenic pain induced by intraplantar injection of capsaicin (1.6 μg/paw), glutamate (10 μmol/paw) and phorbol 12-myristate 13-acetate (1.6 μg/paw) with calculated mean ID₅₀ of 128.8 mg/kg (118.6-139.9 mg/kg), 124.8 mg/kg (111.4-139.7 mg/kg) and 40.29 (35.39-45.86) mg/kg, respectively. It was also demonstrated that pretreatment with l-arginine (100 mg/kg, i.p.), a nitric oxide precursor significantly reversed antinociception produced by EOZZ suggesting the involvement of l-arginine/nitric oxide pathway. In addition, methylene blue (20 mg/kg, i.p.) significantly enhanced antinociception produced by EOZZ. Administration of glibenclamide (10 mg/kg, i.p.), an ATP-sensitive K⁺ channel antagonist significantly reversed antinociceptive activity induced by EOZZ.

Conclusion

Together, the present results suggested that EOZZ-induced antinociceptive activity was possibly related to its ability to inhibit glutamatergic system, TRPV1 receptors as well as through activation of l-arginine/nitric oxide/cGMP/protein kinase C/ATP-sensitive K⁺ channel pathway.     

Bronchodilatory effect of Acorus calamus (Linn.) is mediated through multiple pathways

Aim of the study

This study was undertaken to provide a pharmacological basis for traditional use of Acorus calamus in airways disorders.

Materials and methods

Isolated guinea-pig trachea and atria were suspended in organ baths bubbled with carbogen and mechanisms were found using different parameters.

Results

In isolated guinea-pig tracheal segments, crude extract of Acorus calamus was more effective than carbachol in causing relaxation of high K⁺ (80 mM) precontractions, similar to verapamil, suggesting blockade of calcium channels. The n-hexane fraction was equipotent against both precontractions, similar to papaverine, while ethylacetate fraction was more potent against carbachol precontractions but had a negligible dilator effect against K⁺, similar to atropine and or rolipram. Pretreatment of tracheal preparations with n-hexane or ethylacetate fractions potentiated isoprenaline-induced inhibitory concentration-response curves, similar to papaverine or rolipram. Pretreatment of tracheal preparations with ethylacetate fraction caused a rightward parallel shift in carbachol response curve at lower concentration (0.003 mg/mL) similar to atropine and a non-parallel shift at higher concentrations (0.01 mg/mL), with reduction of maximum response, similar to rolipram. In isolated guinea-pig atrial preparations, crude extracts, its fractions and papaverine inhibited force and rate of contractions at higher concentrations than the smooth muscle while verapamil was equipotent.

Conclusion

These data indicate the presence of unique combination of airways relaxant constituents in crude extract of Acorus calamus, a papaverine-like dual inhibitor of calcium channels and phosphodiesterase in n-hexane fraction and a novel combination of anticholinergic, rolipram-like phosphodiesterase4 inhibitor in ethylacetate fraction and associated cardiac depressant effect, provide a pharmacological basis for traditional use of Acorus calamus in disorders of airways.     

Increased sexual motivation in female rats treated with Humulus lupulus L. extract

Aim of the study

To evaluate the influence of Humulus lupulus extract on sexual behavior in female rats.

Materials and methods

Ovariectomized rats hormonally primed with estradiol benzoate (1.5 μg/rat) and progesterone (500 μg/rat) were acutely treated by oral gavage with Humulus lupulus extract dosed at 5, 10 and 25 mg/kg and then tested for partner preference and sexual receptivity.

Results

The administration of Humulus lupulus extract at the highest dose significantly increased the preference for the stimulus male during the partner preference test and the number of proceptive behaviors during the receptivity test, without affecting the lordosis response.

Conclusions

Humulus lupulus extract increased sexual motivation in hormone-primed female rats.     

Characterisation of the anti-inflammatory and antinociceptive activities and the mechanism of the action of Lippia gracilis essential oil

Aim of the study

Lubricating gut pill (LGP), a traditional Chinese formula, had been conformed to improve the loperamide-induced rat constipation by stimulation of Cl⁻ secretion, but its mechanism has not been fully explored. Thus, the purpose of this study was to identify the action sites of LGP-stimulated Cl⁻ secretion across rat distal colonic mucosa.

Materials and methods

Rat distal colonic mucosa was mounted in Ussing chambers and short circuit current (I_SC), apical Cl⁻ current and basolateral K⁺ current were recorded. Intracellular cyclic adenosine monophosphate (cAMP) content and protein kinase A (PKA) activity were determined with ELISA kit and the non-radioactive PepTag test, respectively.

Results

LGP at 800 μg/ml elicited a sustained increase in Cl⁻ secretory response, which was inhibited by CFTR_inh172, a cystic fibrosis transmembrane conductance regulator (CFTR) inhibitor. Permeabilizing apical membrane with nystatin revealed that LGP-stimulated basolateral K⁺ current was significantly inhibited by KCNQ1 K⁺ channel inhibitor chromanol 293B. LGP-stimulated I_SC was markedly reduced by pretreatment with cis-N-[2-phenylcyclopentyl]-azacyclotridec-1-en-2amine (MDL-12,330A) and N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H-89), but not with inhibitors of Ca²⁺-dependent signaling pathway. Treatment of tissue with LGP resulted in an increase in intracellular cAMP level and the activation in protein kinase A. The E-prostanoid₄ (EP)₄ receptor antagonist L-161,982 completely eliminated LGP-induced response.

Conclusions

The results showed that LGP enhances Cl⁻ and fluid secretion via prostanoid receptor signaling and also cAMP and protein kinase A pathway, subsequently triggering the activation of apical Cl⁻ channels mostly CFTR and basolateral cAMP-dependent K⁺ channel.     

Sida rhomboidea.Roxb leaf extract ameliorates gentamicin induced nephrotoxicity and renal dysfunction in rats

Ethnopharmacological relevance

Sida rhomboidea.Roxb (SR) known as “Mahabala” in Ayurveda and marketed as “Shahadeyi” is used in ethnomedicine to treat ailments such as dysuria and urinary disorders.

Aim of the study

To evaluate nephroprotective potential of SR against gentamicin (GM) induced nephrotoxicity and renal dysfunction.

Materials and methods

Nephrotoxicity was induced in rats with GM (100 mg/kg bodyweight (i.p.) for 8 days) and were treated with SR extract (200 and 400 mg/kg bodyweight (p.o.) for 8 days) or 0.5% carboxymethyl cellulose (vehicle). Plasma and urine urea and creatinine, renal enzymatic and non-enzymatic antioxidants along with lipid peroxidation were evaluated in various experimental groups.

Results

GM treatment induced significant elevation (p < 0.05) in plasma and urine urea, creatinine, renal lipid peroxidation along with significant decrement (p < 0.05) in renal enzymatic and non-enzymatic antioxidants. SR treatment to GM treated rats (GM + SR) recorded significant decrement (p < 0.05) in plasma and urine urea and creatinine, renal lipid peroxidation along with significant increment (p < 0.05) in renal enzymatic and non-enzymatic antioxidants.

Conclusion

SR leaf extract ameliorates GM induced nephrotoxicity and renal dysfunction and thus validates its ethnomedicinal use.     

Diuretic and potassium-sparing effect of isoquercitrin-an active flavonoid of Tropaeolum majus L

Aim of the study

Previous studies have shown that the extracts obtained from Tropaeolum majus L. exhibit pronounced diuretic effects supporting the ethnopharmacological use of this plant as diuretic. In the present work, phytochemical investigation, guided by bio-assay in spontaneously hypertensive rats (SHR), was carried out in order to identify the compounds responsible for diuretic action.

Material and methods

Chromatographic fractionation of the hydroethanolic extract yielded an active fraction (TMLR) rich in isoquercitrin. TMLR (25-100 mg/kg) and isoquercitrin (5-10 mg/kg), as well the reference drug hydrochlorothiazide (10 mg/kg) were orally administered in a single dose or daily for 7 days to SHR. The urine excretion rate, pH, density, conductivity and content of sodium (Na⁺) and potassium (K⁺) electrolytes were measured in the urine of saline-loaded animals.

Results

The urinary excretion rate was dose-dependently increased in both TMLR and isoquercitrin groups, as well as Na⁺. Despite the changes in urinary excretion of electrolytes, the plasmatic levels of Na⁺ and K⁺ had not been changed. In addition, we did not find any evidence of renal toxicity or other adverse effects in these animals, even after prolonged treatment with TMLR or isoquercitrin.

Conclusion

This research supports and extends the ethnomedicinal use of T. majus as diuretic. This activity seems to be associated to the presence of the flavonol isoquercitrin.     

Inonotus obliquus extracts suppress antigen-specific IgE production through the modulation of Th1/Th2 cytokines in ovalbumin-sensitized mice

Ethnopharmacological relevance

Chaga mushroom (Inonotus obliquus, IO) has been used as a folk remedy for cancer, digestive system diseases, and other illnesses in Russia and Eastern Europe.

Aim of the study

In the present study, we investigated the immunomodulating effects of IO through in vivo and ex vivo studies.

Materials and methods

Serum immunoglobulins (IgE, IgG₁, and IgG_2a) and cytokines (interleukin (IL)-4, interferon (IFN)-γ, and IL-2) were measured in concanavalin A (ConA)-stimulated splenocytes and CD4⁺ T cells. The nitric oxide (NO) secretion of lipopolysaccharide (LPS)-stimulated peritoneal macrophages was also measured after oral administration of 50, 100, or 200 mg kg⁻¹ d⁻¹ IO hot water extract (IOE) to ovalbumin (OVA)-sensitized BALB/c mice.

Results

We found that the OVA-induced increase in serum IgE and IgG_2a was significantly suppressed when IOE was orally administered after the second immunization with OVA. ConA stimulation in spleen cells isolated from OVA-sensitized mice treated with 100 mg kg⁻¹ IOE resulted in a 25.2% decrease in IL-4 production and a 102.4% increase in IFN-γ, compared to the controls. Moreover, IL-4, IFN-γ, and IL-2 were significantly reduced after ConA stimulation in isolated CD4⁺T cells. We also determined that IOE inhibits the secretion of NO from LPS-stimulated peritoneal macrophages ex vivo.

Conclusions

We suggest that IO modulates immune responses through secretion of Th1/Th2 cytokines in immune cells and regulates antigen-specific antibody production.     

Hepatoprotective activity of the edible snail (Bellamia bengalensis) flesh extract in carbon tetrachloride induced hepatotoxicity in rats

Ethnopharmacological relevance

In the folk-traditional medicine, snails were used to purify blood, boost immune system, prevent conjunctivitis and to treat liver problems.

Objectives

To evaluate the hepatoprotective activity of the edible snail (Bellamia bengalensis) flesh extract in male Wistar rats treated with carbon tetrachloride as an hepatotoxicant.

Materials and methods

Live adult Bellamia bengalensis was collected commercially from the Kolkata market. Aqueous flesh extract (BBE) was prepared in 0.9% saline and expressed in terms of wet weight basis. The aqueous flesh extract was administered orally (1, 2 g kg⁻¹ day⁻¹) to male rats for 12 days. Liv52 was used as positive control. 24 h after administration of extract, the rats were given a single oral dose of CCl₄ (1.25 ml kg⁻¹), except vehicle control rats. After 24 h of CCl₄ administration, all the animals were sacrificed to collect the blood and liver tissue.

Results

BBE (1 and 2 g kg⁻¹ day⁻¹, p.o. × 12 days) significantly prevented CCl₄ induced elevation of SGOT, SGPT, γGT, ACP, ALP, bilirubin, LDH and CCl₄ induced decrease in total protein, triglyceride level in male Wistar rats. BBE treated rat liver anti-oxidant parameters (LPO, SOD, GSH, CAT, GPx) were significantly antagonized for the pro-oxidant effect of CCl₄. Histopathological studies also supported the protective effect of BBE.

Conclusion

This study validated the folk and traditional use of snail in liver disorder through CCl₄-induced rat experimental model.