Evaluation of anti-amnesic effect of Conyza bonariensis in rats.

OBJECTIVE: Conyza bonariensis is an ornamental medicinal weed. This experiment was planned to explore the outcome of petroleum ether extract of C. bonariensis (PECB) leaves on scopolamine-induced amnesia in rats. MATERIALS AND METHODS: For impairing memory, 0.4 mg/kg (i. p.) of scopolamine was given. Fifty to 200 mg/kg of PECB was fed orally to rats and 3 mg/kg (i. p.) of tacrine was given as a standard drug. Anti-amnesic property was evaluated in Barnes maze using ANY-maze software. Following a behavioral study, acetylcholinesterase (AChE), ß-amyloid1-41, antioxidant enzymes, and cytokine levels were measured. Furthermore, reverse transcription-polymerase chain reaction was done for expression of the marker genes such as AChE, Nrf2, NF-κB, PP2A, and HO-1, whereas BDNF, TrkB, caspase-3, and Bax were measured by Western blotting. RESULTS: PECB and tacrine significantly improved memory dysfunction by decreasing escape latency in Barnes maze. At the highest dose, treatment with PECB altered the scopolamine-induced hyperactivation of AChE and ß-amyloid1-41 activity. PECB elevated the levels of superoxide dismutase, glutathione, and catalase and decreased lipid peroxidation and nitric oxide dose dependently. PECB attenuated scopolamine-induced increase of tumor necrosis factor-α and interleukin (IL)-1ß concentrations in the hippocampus with reversed diminished IL-10 level toward normal in the brain. Nrf2, HO-1, PP2A, BDNF, and TrkB were significantly upregulated with downregulation of AChE, NF-κB, Tau, Bax, and caspase-3. Different components such as beta-amyrin and alpha-amyrin were isolated from leaves of the plant. CONCLUSION: The results indicated that PECB might be a potential curative drug for the treatment of cognitive impairment.

Zanthoxylum alatum Roxb. seed extract ameliorates stress aggravated DSS-induced ulcerative colitis in mice: Plausible role on NF-κB signaling axis.

ETHNOPHARMACOLOGICAL RELEVANCE: Zanthoxylum alatum (ZA) Roxb (family: Rutaceae) plant has been traditionally used for multiple indications by local healers among different communities of South Asian countries mainly in India and Bangladesh. The extracts of ZA have reported strong anti-inflammatory and anti-oxidant activities, but no scientific report is available on its efficacy in intestinal inflammatory disorders like ulcerative colitis. AIM OF THE STUDY: The overall objective of our study was to evaluate the anti-inflammatory potency of hydro-ethanolic extract of Zanthoxylum alatum seed (ZAHA) using both in-vitro NF-κB-luciferase translocation assay and in-vivo stress aggravated dextran sodium sulfate (DSS)-induced ulcerative colitis model. MATERIALS AND METHODS: The in-vitro anti-inflammatory effect of ZAHA extract was evaluated by luciferase assay in HEK293 cells. Parameters such as body weights, behavioural, colonoscopy, colon lengths and spleen weights were measured and recorded in stress aggravated DSS-induced colitis model in C57BL/6 mice. Biochemical, histological and immunoblot analysis in the colon tissues were determined to prove its anti-inflammatory and anti-oxidant activities. Characterization of the extract was done by LC-MS/MS study. RESULTS: Initial in vitro NF-κB-luciferase translocation assay showed that the hydroalcoholic extract of ZA (ZAHA) showed potent inhibitory activity for NF-κB translocation by TNF-α stimulation and hence this particular extract was further evaluated in stress aggravated DSS-induced ulcerative colitis model in C57BL/6 mice. Treatment of ZAHA for two weeks at a dose of 200 mg/kg significantly ameliorated the stress aggravated DSS-induced colitis in mice. Histological alterations, infiltration of inflammatory cells, and the levels of IL-1ß, IL-6, TNF-α in colon tissue and serum samples were significantly decreased in ZAHA treatment groups compared to the stress aggravated DSS induced colitis animals. Moreover, the protein expressions of p-NF-κB, p-IκBα, p-STAT3, COX-2, and TNF-α were significantly reduced in colon tissues of ZAHA treated groups and also increased anti-oxidant markers like SOD-1, Nrf2 significantly when compared with disease control group. Characterization of the extract further by LC-MS/MS revealed the presence of several active compounds which could be responsible for its anti-inflammatory activity. CONCLUSIONS: Thus from the above findings it can be concluded that ZAHA ameliorates stress aggravated DSS-induced ulcerative colitis due to its anti-inflammatory and anti-oxidant activity.

Effect of seeds of Entada phaseoloides on chronic restrain stress in mice.

BACKGROUND: Entada phaseoloides is a well-known medicinal plant traditionally used in Ayurvedic medicine for centuries. OBJECTIVE: To evaluate the anti-stress activity of seeds of E. phaseoloides in endoplasmic reticulum stress during chronic restrain stress in mice, based on our preliminary screening. MATERIALS AND METHODS: Mice (n = 6/group) were restrained daily for 6 h in 50 ml polystyrene tubes for 28 days. Methanolic extract of E. phaseoloides (MEEP) (100 and 200 mg/kg, p.o.) and standard drug, imipramine (10 mg/kg i.p.) were administered daily 45 min prior to restrain from day 22-28. Then, forced swim test (FST) was performed to assess despair behavior. Lipid peroxidation (LPO) and antioxidant enzymes Reduced glutathione (GSH), Superoxide dismutase (SOD) were measured in the hippocampus of mice. 78 kDa Glucose-regulated Protein, 94 kDa Glucose-regulated Protein, C/EBP homologous protein, Caspase-12 expression were quantified by Real Time PCR. RESULTS: MEEP significantly reduced the immobility time in FST (P < 0.001). Significant reduction of LPO (P < 0.05) level and restored antioxidant enzymes viz. GSH (P < 0.001) and SOD towards vehicle control group were observed. Down-regulation of genes GRP 78, GRP 94 (P < 0.001), CHOP and Caspase-12 (P < 0.001) as compared to the chronic restrain stress group was evident, which were upregulated following treatment. Isolation of the active components of the seeds revealed the presence of Oleic acid (1), Entadamide A (2), Entadamide A-beta-d-glucopyranoside (3) and 1-O-protocatechuoyl-ß-d-glucose. CONCLUSION: MEEP altered endoplasmic reticulum stress in chronic restrain stressed mice; however, as an antidepressant it showed a weaker response.

Zanthoxylum alatum abrogates lipopolysaccharide-induced depression-like behaviours in mice by modulating neuroinflammation and monoamine neurotransmitters in the hippocampus.

CONTEXT: Depression is an inflammatory, commonly occurring and lethal psychiatric disorder having high lifetime prevalence. Zanthoxylum alatum Roxb. (Rutaceae), commonly called Timur, has high medicinal value and is used ethnomedicinally for the treatment of various diseases. OBJECTIVE: To evaluate the effect of hexane extract of Z. alatum seeds (ZAHE) on lipopolysaccharide (LPS)-induced depression-like behaviour in Swiss albino mice. MATERIALS AND METHODS: Mice were treated with ZAHE (100 and 200 mg/kg, p.o.) and imipramine (10 mg/kg injected i.p.) for 14 days. On 14th day of the treatment, depression-like behaviour was induced by LPS (0.83 mg/kg injected i.p.) and after 24 h of LPS administration, it was assessed by measuring behavioural parameters and biochemical estimations. RESULTS: Behavioural tests, including the open field test, forced swimming test, tail suspension test and sucrose preference test revealed that ZAHE (100 and 200 mg/kg, p.o.) and imipramine (10 mg/kg injected i.p.) alleviated the depression symptoms of LPS-induced mice. Moreover, ZAHE treatments reversed the LPS-induced alterations in the concentrations of norepinephrine and serotonin (5-HT) and inhibited the expression of brain-derived neurotrophic factor, pro-inflammatory cytokines and oxido-nitrosative stress in the mice. Acute toxicity was calculated to be LD50 > 2500 mg/kg. DISCUSSION AND CONCLUSIONS: This study showed that LPS-induced depression in mice was significantly prevented by ZAHE at both the dosages. In conclusion, ZAHE exhibited an antidepressant activity by altering monoaminergic neurotransmitters in the brain combined with its anti-inflammatory potential. Thus, it could be an effective therapeutic against inflammation-induced depression and other brain disorders.

Role of Elsholtzia communis in counteracting stress by modulating expression of hspa14, C/EBP homologous protein, nuclear factor (erythroid-derived 2)-like-2 factor, Caspase-3, and brain-derived neurotrophic factor in rat hippocampus.

OBJECTIVE: Elsholtzia communis (Collett and Hemsl.) Diels has been widely distributed and is reported for many therapeutic effects. The present study aims to investigate the antistress activity of the leaf extract and its possible molecular mechanism. MATERIALS AND METHODS: Hydroethanolic extract of leaves of E. communis (100 and 200 mg/kg, p.o.) were administered for 7 days to stress-induced male Wistar rats. The experimental animals were divided into five groups (n = 6). The mRNA/protein profile of few stress responsive chaperones (hspa14), endoplasmic reticulum stress markers (C/EBP homologous protein [CHOP]), antioxidant regulating genes (nuclear factor (erythroid-derived 2)-like-2 factor [Nrf2]), apoptotic factors (Caspase-3) in rat hippocampus were studied by polymerase chain reaction and immunoblotting. RESULTS: The stress-related genes such as hspa14, CHOP, antioxidant gene Nrf2, apoptotic gene Caspase-3 which were overexpressed in the stress control group were significantly suppressed following administration of the extract at both the doses and the standard drug Ginseng. Likewise, brain-derived neurotrophic factor which is closely related with stress, was downregulated in the stress control group, was found to be upregulated following treatment with the extract and the standard drug Ginseng. CONCLUSION: Our findings clearly indicate that E. communis was able to counteract stress. Hence, it has the potential to develop as adaptogen and also as a replacement/substitute of the popularly used drug, Ginseng or Ashwagandha, which is on the verge of extinction or becoming endemic due to overuse.

Anticholinergic, antihistaminic, and antiserotonergic activity of n-hexane extract of Zanthoxylum alatum seeds on isolated tissue preparations: An ex vivo study.

OBJECTIVES: The aim of this study was to evaluate anticholinergic, antihistaminic, and antiserotonergic activity of the n-hexane extract of the seeds of Zanthoxylum alatum (ZAHE) on isolated ileum of rat and guinea pig and fundus of rat. MATERIALS AND METHODS: ZAHE was prepared using soxhlet extraction and cumulative concentration response curves were constructed using various doses on the tissues for acetylcholine (ACh), 5-hydroxytryptamine (5-HT), and histamine with or without n-hexane extract. Atropine, ketanserin, and pheniramine maleate were used as antagonists for ACh, serotonin, and histamine, respectively. RESULTS: ZAHE-induced concentration-dependent inhibition of isolated ileum and fundus in rat and ileum of guinea pig. The half maximal effective concentration (EC50) of ACh in the presence of atropine (10-6 M; P < 0.05) and ZAHE (1000 µg/ml; P < 0.01) was significantly higher than EC50of ACh alone. The EC50of 5-HT in the presence of ketanserin (10-5 M; P < 0.01) and ZAHE (1000 µg/ml; P < 0.05) was higher than EC50of 5-HT alone. Similarly, the EC50of histamine in the presence of pheniramine maleate (10-6 M; P < 0.01) and ZAHE (300 µg/ml; P < 0.01 and 1000 µg/ml; P < 0.05) was also significantly higher than EC50of histamine alone. CONCLUSION: From the study, it was observed that ZAHE shows significant anticholinergic, antiserotonergic, and antihistaminic activity. The study provides sufficient evidence that the seeds can be used in gastric disorders, cough, chest infection, etc., as per folklore claims.

Hematobiochemical and pathological alterations due to chronic chlorpyrifos intoxication in indigenous chicken.

OBJECTIVE: The present study investigates the effect of oral administration of chlorpyrifos (CPF) in indigenous chicken. MATERIALS AND METHODS: The birds were divided into two groups I and II. Group I served as control and group II was treated with CPF (0.36 mg/kg) orally daily up to 12 weeks. Blood samples were assayed for hemoglobin (Hb), total erythrocyte count (TEC), total leukocyte count (TLC), differential leukocyte count, and biochemical constituents like alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholinesterase (CHE), total protein and uric acid. Representative pieces of tissues from liver and kidney were collected weekly for histopathological examination. RESULTS: A significant (P < 0.01) increase of Hb, TEC, TLC, and heterophil percent and decrease of lymphocyte percent was observed. Serum ALP, AST, ALT, and uric acid increased significantly and CHE values decreased significantly in CPF treated birds. The protein level remained similar. Uric acid level was found to be increased significantly in the treated group. The results indicate that chronic CPF intoxication produces hematological, biochemical, and pathological changes in treated birds.

Influence of Alternanthera brasiliana (L.) Kuntze on Altered Antioxidant Enzyme Profile during Cutaneous Wound Healing in Immunocompromised Rats.

Alternanthera brasiliana (L.) Kuntze (Amaranthaceae) is a herbaceous plant used against inflammation, cough, and diarrhea in Brazilian popular medicine. In our preliminary study, promising wound healing activity of methanol extract of leaves of A. brasiliana (MEAB) was observed in normal excision and incision wound models. Therefore, the present study was designed to investigate the wound healing activity along with the antioxidant enzyme profile during cutaneous excision immunocompromised wound after topical application of 5% w/w ointment of MEAB in rats. Immunocompromised state was induced by pretreatment with hydrocortisone (HC) at 40 mg/kg body weight (i.m.) in male rats. Following one-week pretreatment with HC, wounds were created. The vehicle, 5% (w/w) ointment of MEAB, or standard drug (Himax) was applied topically twice daily. Healing potential was evaluated by the rate of wound contraction, estimation of enzymatic and nonenzymatic antioxidants like catalase, SOD, GSH, protein, vitamin C, and hydroxyproline content, which was supported by histopathological study on the 8th day following wounding. There was significant increase in the enzymatic and nonenzymatic antioxidant parameters in the extract-reated group as compared to control group. Histopathological study revealed collagen deposition, fibroblast proliferation, angiogenesis, and development of basement membrane in A. brasiliana group. The results of the present investigation revealed significant wound healing activity of MEAB.

In vivo wound-healing efficacy and antioxidant activity of Achyranthes aspera in experimental burns.

CONTEXT: The floral richness of the North-East Indian region cannot be neglected in context to its medicinal importance. Achyranthes aspera Linn. (Amaranthaceae; Prickly Chaff flower) is an indigenous plant species of this region. Although the local traditional healers have ethnomedical knowledge on the use of this plant, there is no scientific study on wound-healing activity of this plant. OBJECTIVE: The healing efficacy of methanol leaf extract of A. aspera (MEAA) in granulation tissue of burn wound and its antioxidant activity are investigated. MATERIALS AND METHODS: Methanol extract of leaves of A. aspera was used for compounding 5% (w/w) ointment, which was applied topically twice daily in experimental burn wound in rats. Healing potential was assessed by rate of wound contraction, antioxidant and biochemical assay which was supported by gelatin zymography and histopathology. RESULTS: In the present study, 5% ointment of A. aspera showed significant (p < 0.05) wound healing, which was evident by wound contraction, elevation of various antioxidant enzymes viz. SOD, catalase, vitamin C and prohealing and biochemical parameters like hydroxyproline and protein content than the control animals. Up-regulated expression of matrix metalloproteinases (MMP-2 and 9) was also observed by gelatin zymography. Histopathological examination of the granulation tissues in the A. aspera-treated animals showed collagen deposition, fibroblast proliferation and formation of epidermis. DISCUSSION AND CONCLUSION: The methanol leaf extract of A. aspera showed excellent wound-healing activities which has great potential for development of plant-based product.

Healing efficacy of methanol extract of leaves of Alternanthera brasiliana Kuntze in aged wound model.

The methanol extract of Alternanthera brasiliana Kuntze (Family: Amaranthaceae) leaf was investigated for its wound healing effect by excision wound model (in vivo) in aged Sprague Dawley rats. In excision wound model, compared to the control group, percent contraction of wound was significantly (P < 0.01) higher in A. brasiliana-treated group (5% w/w ointment). The collagen, elastin, and hydroxyproline contents of the granulation tissue of A. brasiliana-treated group increased significantly (P < 0.01) compared to the control group, indicating better wound healing activity of the test plant. These findings were also confirmed by histopathological examination. The results suggested that methanol extract of A. brasiliana possesses significant wound healing potential in aged animal wound model.

Analgesic and anti-nociceptive activity of hydroethanolic extract of Drymaria cordata Willd.

OBJECTIVES: To study the analgesic and anti-nociceptive activity of hydroethanolic extract of Drymaria cordata Willd. MATERIALS AND METHODS: Wistar rats and Swiss albino mice were used for studying analgesic and anti-nociceptive activity of Drymaria cordata hydroethanolic extract (DCHE) at doses 50, 100 and 200 mg/kg p.o. Various models viz. acetic acid induced writhing model (female mice), Eddy's hot plate (mice) and tail flick model (rat) for analgesic study and formalin-induced paw licking model (mice) were used for anti-nociceptive study. RESULTS: In acetic acid induced writhing model, effect of DCHE was better than the standard drug- indomethacin 10 mg/kg (p.o.). In the hot plate model, the maximum effect was observed at 60 min at a dose of 200 mg/kg p.o., which was higher than the standard drug morphine sulfate (1.5 mg/kg i.p.), whereas in the tail flick model, effect was comparable with morphine sulfate. In formalin-induced paw licking model, administration of DCHE completely abolished the early phase at 100 and 200 mg/kg p.o. and in the late phase, the effect of DCHE (200 mg/kg p.o.) was higher than indomethacin (10 mg/kg p.o.). CONCLUSION: DCHE was effective in both non-narcotic and narcotic models of nociception, suggesting its possible action via peripheral and central mechanism. It also abolished the early phase in formalin-induced paw licking model, suggesting complete inactivation of C-fiber at higher dose. The activity can be attributed to the phyto-constituents viz tannins, diterpenes, triterpenes and steroids present in the DCHE extract. In conclusion, DCHE can be developed as a potent analgesic and anti-nociceptive agent in future.

Studies on the anti-inflammatory properties of Plantago erosa leaf extract in rodents.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves of Plantago erosa ex Roxb are used traditionally in Northeast India in different illnesses which include wounds, cuts, bruises, insect bites, poison-ivy rashes, minor sores and snakebite, etc. AIM OF THE STUDY: Plantago erosa is one of the commonly used medicinal plants in various inflammatory conditions in this region; however, due to paucity of scientific literature on its anti-inflammatory property, the present study was aimed at evaluating its anti-inflammatory activity in the leaves using in vivo models of inflammation. MATERIALS AND METHODS: Different models like carageenan induced paw edema in rat and mice, formalin induced paw licking in rats and cotton pellet induced granuloma in rats were used for studying the anti-inflammatory activity in methanol extract of Plantago erosa (PEME) leaves. RESULTS: The PEME at the oral doses from 300 to 600 mg/kg showed anti-inflammatory activity in various models. The extract (PEME) reduced carageenan induced paw edema in rat and mice, inhibited the formation of granulomatous tissue in cotton pellet induced granuloma after treatment and also decreased the reaction time in both early and late phases in formalin induced paw licking in rats. CONCLUSION: The study evidently confirmed anti-inflammatory activity of PEME and thus supported the traditional claim. The anti-inflammatory activity could be attributed to the phytoconstituent (flavonoids, alkaloids and steroid) present in the methanol extract of the plant.