Pharmacological evaluation of cardioprotective potential of Berberis orthobotrys Bien.ex Aitch.

The resurgence of scrutiny in plant-based medicine is mainly due to the current widespread belief that "green medicine" is safe and more dependable than the expensive synthetic drugs. The current study was focused to evaluate the anti-myocardial ischemic potential of Berberis orthobotrys Bien ex Aitch against chemically induced myocardial ischemia in animal models. Myocardial ischemia was instigated in Sprague Dawley rats of either sex (250-450g) by administration of Isoproterenol (ISO) and doxorubicin (DOX) at doses of 25mg/kg b.w and 15mg/kg b.w. respectively. The protective effect of the plant extract was explored by pretreating a group of animals with aqueous methanolic extract of Berberis orthobotrys roots at a dose of 50mg/kg b.w. (orally) for 10 days in ISO-ischemic model while for doxorubicin ischemic model; the study was conducted for 14 days. The findings of the study revealed that serum levels of cardiac marker enzymes were significantly increased (p<0.0001) followed by the administration of Isoproterenol and doxorubicin whereas the pretreatment with aqueous methanolic plant extract had significantly (p<0.0001) prevented the rise in the same, as compared to both intoxicated groups. The statistical analysis of the study led to the conclusion that Berberis orthobotrys possesses cardio protective potential against chemically induced myocardial ischemia.

Pharmacological exploration of anti-arthritic potential of terbutaline through in-vitro and in-vivo experimental models.

Terbutaline have been reported to have anti-inflammatory activity. Present study aimed to check the anti-arthritic activity of terbutaline. The drug was tested using in vitro models (bovine serum albumin denaturation, egg albumin denaturation and HRBC membrane stabilization) and in vivo (formaldehyde induced arthritis). Results of bovine serum albumin denaturation assay illustrated that terbutaline inhibited 89.54±0.46% denaturation at 6400µg/ml concentration. Terbutaline resulted in dose dependent impediment of protein denaturation in egg albumin denaturation assay with 74.40±0.72% inhibition at concentration of 6400µg/ml. Terbutaline also showed protection of HRBC membrane against hypotonic stress in a dose dependent manner, with maximum 76.45±0.62% prevention at 6400µg/ml concentration. Results of formaldehyde induced arthritis model showed that paw volume was significantly declined by terbutaline with maximum percentage inhibition at 10th day of study period which implies immune inhibitory potential of terbutaline. Findings of present study concluded that terbutaline has arthritis reducing potential possible through inhibitory effects on synthesis and release of inflammatory mediators as well as limiting the formation of autoantigen. Thus, terbutaline might be the potential candidate for use in treatment of arthritis.

Cardioprotective potential of Thymus linearis Benth.

In developing countries, myocardial ischemia and the resulting impairments in heart function are the leading cause of illness and mortality. Thymus linearis Benth has been used as an antibiotic, antioxidant, and antihypertensive agent for centuries. The goal of this investigation was to see if Thymus linearis could protect isoproterenol and doxorubicin-induced myocardial ischemia in vivo at doses of 25 mg/kg s.c. and 15 mg/kg i.p., respectively. The level of cardiac enzymes (CK-MB, LDH, and AST) in the serum isolated from the experimental animal's blood was used to determine myocardial ischemia. The anti-ischemic potential was assessed by comparing the levels of the aforementioned cardiac biomarkers in the intoxicated and treated animal groups. The study found substantial increase (p0.0001) in the serum levels of CK-MB, LDH, AST when compared to intoxicated groups, while pretreatment of animals with crude extract of Thymus linearis significantly reduced the rise in serum cardiac indicators. The findings of the study indicated that the aqueous methanolic Thymus linearis crude extract has cardioprotective potential against Isoproterenol and Doxorubicin-induced cardiac necrosis in rats.

Therapeutic appraisal of ephedrine against rheumatoid arthritis: A new indication.

Ephedra, natural flora has been used traditionally to treat rheumatism since decades. The scientific evidence of anti-rheumatic effect of this plant has also been reported. But the anti-rheumatic activity of major constituent of this plant (ephedrine) has not been evaluated. Based on this, the current study was aimed to assess anti-arthritic activity of ephedrine by using in vitro and in vivo approaches. Correspondingly, enzyme linked immunosorbent assay was performed for the estimation of prostaglandins E2 (PGE2) and tumor necrosis factor-α (TNF-α) in serum of formaldehyde-induced arthritic animals. The results elaborated significant reduction in albumin denaturation and remarkable progress on stabilization of red blood cells outer membrane at higher concentration during in vitro experiments. The ephedrine (40mg/kg) revealed noteworthy (p<0.001) inhibition in paw swelling in animals intoxicated with albumin as well as formaldehyde as compared to animals of control group by in vivo results. In this assay, ephedrine (20 & 40 mg/kg orally) significantly suppressed the level of these inflammatory markers (PGE2 & TNF-α). Ephedrine exhibited anti-arthritic effect by decreasing pro-inflammatory cytokines (PGE2 & TNF-α). This experimental work pharmacologically supports the use of ephedrine as anti-rheumatic drug but limited to evaluate in immunological arthritic model.

Ethyl-acetate extract of tara mira (Eruca sativa) alleviates the inflammation and rheumatoid arthritis in rats.

Eruca sativa, member of family Brassicaceae, was evaluated for its anti-arthritic potential. Both in vitro and in vivo models were used to bring out a safe, effective and economical remedy. In vitro tests included egg albumin denaturation suppression, bovine serum albumin assay and human red blood cells maintenance assay. While in vivo formaldehyde-induced arthritic model was initiated to check effect on paw volume. Similarly, carrageenan produced inflammation was applied to check anti-inflammatory ability of the plant. Acute toxicity studies showed safety margin at 2000mg/kg. The plant showed concentration dependent denaturation protection and membrane stability in vitro assays. Likewise, the carrageenan and formaldehyde investigations revealed visible paw volume reduction in dose attributed manner, with maximum outcome at dose of 500mg/kg. Hence, it may be established on the ground of presented results that ethyl-acetate extract of Eruca sativa has significant anti-inflammatory and anti-arthritic effects and may be considered for further research to reveal the core mechanism.

Evaluation of phytochemical, anti-oxidant and cardiac depressant effect of Rumex dentatus by using Langendorff's isolated heart apparatus.

Rumex dentatus has been used traditionally for ailment of cardiovascular diseases. The aim of the present study was to assess cardiovascular effects in isolated perfused rabbit heart. Aqueous and n-butanolic fractions were assessed for their effect on perfusion pressure (PP), force of contraction (FC) and heart rate (HR) of rabbit heart using Langendorff's method. The possible mechanisms of action of extracts/fraction were assessed with and without application of different agonist/antagonist. Phytochemical, toxicity and anti-oxidant activities were also determined. Both extracts at 1mg/mL dose produced a highly significant decrease in FC and HR but PP remained unchanged. Moreover, aqueous fraction of Rumex dentatus at 0.001mg/mL dose produced a highly significant decrease in FC and HR but no significant change in PP was observed. Atropine 10-5 M did not inhibit the cardiac depressant response of both fractions. Furthermore, both fractions blocked the positive ionotropic and chronotropic effects of adrenaline and calcium chloride. Phytochemical studies have shown the presence of some phytochemicals. Acute and sub-chronic toxicity studies demonstrated that test extracts are safe and produced no significant changes in haematological and biochemical parameters. Crude extract showed significant antioxidant activity like ascorbic acid. This study revealed that this plant have good cardiac depressant effect.

Antihypertensive and safety studies of Cydonia oblonga M.

Antihypertensive studies on aqueous-methanolic extract prepared from seeds of Cydonia oblonga M. were carried out. The test extract in 200, 400 and 600 mg/kg doses was investigated in normotensive, high cholesterol and glucose fed hypertensive rats through non-invasive blood pressure measuring technique. Acute and sub-chronic toxicity studies were conducted in mice and rats, respectively. The test extract significantly decreased dose dependently the systolic, diastolic and mean arterial pressures. The test extract in 600mg/kg dose produced maximum effect and prevented rise in blood pressure of high cholesterol diet and glucose fed rats as compare to control in 21 days studies. The extract was found safe up to 4g/kg dose in mice. In sub-chronic toxicity study, no significant alteration in blood chemistry of extract treated rats was observed except reduction in the low density cholesterol levels. It is concluded that Cydonia oblonga seeds extract possess antihypertensive effect which supports its use in folklore.

Appraisal of hypotensive effect of natural monoterpene: Bornyl acetate and in-silico molecular docking analysis.

The modern trend of research is highly focused on finding new bioactive molecules from medicinal plants. As a functional bicyclic monoterpene, Bornyl acetate (BA) has displayed antioxidant and anti-inflammatory properties in different types of cells and tissues. The purpose of this research was to evaluate the probable hypotensive effect of BA, an underlying mechanism(s) backboned by in-silico studies. Mean arterial pressure and heart rate were recorded via invasive blood pressure measuring technique in normotensive Sprague-Dawley rats following the administration of BA (1-80mg/kg). Docking studies were carried out with various targets involved in the pathophysiology of hypertension.RO5 and ADMET properties were also evaluated. In the current study dose-dependent reduction in systolic, diastolic and mean arterial pressure was observed. Pretreatment with atropine and captopril significantly (p<0.001) reduced the hypotensive effect produced by BA. On the other hand docking studies showed pronounced interactions with M2 mAch receptor in an agonistic way and ACE protein in an antagonistic way. BA justified all cut-off limits of RO5 and had an acceptable predicted computational toxicity profile. Results postulate that dose-dependent hypotensive effect of BA is mediated through the muscarinic pathway and ACE inhibitory activity corresponding well with findings of in-silico studies.

Endothelium independent calcium channel blocking pathways mediate antihypertensive effect of morus nigra.

Present study was conducted to validate the folkloric claims of morus nigra l. (moraceae) using invasive blood pressure measuring and ex vivo vasorelaxant experimental techniques. Intravenous administration of mn. Aq in 0.01-30 mg/kg doses caused significant decrease in mean arterial pressure and heart rate in fructose-induced hypertensive rats. It also showed relaxation in high k+ [80 mm] and pe (1µm) mediated aortic contraction with ec50 1.25 and 3.72mg/ml values, respectively. Vaso-relaxant effect of mn.aq was partially blocked in presence of l-name with ec50, 5.32mg/ml value, but showed concentration dependent significant inhibition of ligand gated and voltage gated ca+2 channels and intracellular ca+2 release, similar to verapamil. Findings of current study designate that aqueous fraction of m. Nigra possesses antihypertensive activity with concentration-dependent vaso-relaxant effect predominantly mediated through endothelial-independent calcium channel blocking pathways accompanied by partial involvement of endothelium-dependent nos mediated relaxation.

Cardiotonic potential of Carissa opaca Stapf ex Haines: A study on isolated rabbit heart.

Carissa opaca (C.O) is a wild shrub, belonging to the family Apocynaceae. The medicinal virtues of this plant have long been known. The present study demonstrates the effects of aqueous-methanolic extract and various fractions (n-butanolic and aqueous) of Carissa opaca on cardiovascular parameters. The perfusion pressure (PP), force of contraction (FC) and heart rate (HR) were assessed on isolated heart of rabbit using Langendroff's technique for crude extract and fractions of C.O, followed by the elucidation of the mechanism of action after estimating toxicity of the plant. Negative ionotropic and positive chronotropic effects, with an increase in PP in isolated perfused rabbit heart were observed the with plant extract and fractions. The aqueous-methanolic extract exhibited maximum response at 1mg/ml while the n-butanolic and aqueous fractions showed a maximum response at 1mg/ml and 10µg/ml respectively. Both fractions produced the same response when treated with atropine (10-5 M), however the actions of adrenaline (10-5 M) and calcium chloride (10-5 M) remained unblocked. Acute toxicity studies indicated that the plant was safe up to 2000 mg/kg and sub-chronic studies demonstrated that no significant change in haematological and biochemical parameters observed. In conclusion, this study supports the folkloric claim of C.O extract.

Toxicity studies and anti-arthritic effect of mandelic acid (2-hydroxy -2-phenyl acetic acid) using in vitro and in vivo techniques.

The major concern to search for new anti-arthritic drugs is primarily to prevent systemic complications and to maintain quality of life. As these drugs are prescribed for long duration so the objective is to ensure their safety in terms of toxicity. By keeping in view this concept, the present study was investigated to determine new anti-arthritic potential using in-vitro and in-vivo methods. The in-vitro tests comprised of protein denaturation (BSA and egg albumin) and Human Red Blood cell (HRBC) membrane stabilization assays at 50-6400µg/mL, for in-vivo testing, formaldehyde-induced arthritic rats were treated with 40, 80 and 160mg/kg mandelic acid. Mandelic acid (MA) inhibited the protein denaturation and stabilized the membrane of HRBC in a concentration dependent manner. Likewise, mandelic acid exhibited dose dependent reduction in paw volume induced by formaldehyde. For acute and sub-acute treatment, MA did not show any sign of toxicity and mortality in each rat and LD50 might be greater than 2000mg/kg. In addition, histopathological assessment presented slight increased interstitial spaces in the kidney, disorganization of glomerulus, dilated sinusoids at highest dose 800mg/kg which were not observed in sub-chronic therapy. Hence, these results conclude that mandelic acid has the potential to treat rheumatoid arthritis with observed no significant signs of toxicity and should be tested further to determine anti-arthritic mechanism of drug action at cellular level.

Evaluation of anti-inflammatory and analgesic activities of aqueous methanolic extract of Ranunculus muricatus in albino mice.

Ranunculus muricatus (Ranunculaceae) is commonly used by inhabitants of Pakistan for the treatment of gout and rheumatism, both of which are inflammatory disorders. The present study attempts to evaluate the anti-inflammatory and analgesic activities of aqueous methanolic extract of R. muricatus in mice. The plant extract at doses of 50, 100 and 150 mg/kg was tested for anti-inflammatory activity against carrageenan and egg albumin induced paw edema in mice and analgesic activity was appraised against acetic acid induced writhing and formalin induced paw licking in mice models. The results designate that extract at the highest dose of 150 mg/kg significantly (p<0.001) and dose dependently inhibited carrageenan induced and egg albumin induced paw edema. Similarly, extract at the same dose of 150 mg/kg showed potent and dose dependent (p<0.001) suppression of formalin induced paw licking and abdominal constrictions / stretching of hind limbs induced by acetic acid. The anti-inflammatory and analgesic activity of plant extract was comparable to standard drug ibuprofen in all models. This study thus supports the use of R. muricatus in traditional medicine for conditions associated with inflammation and analgesia which might be attributed to its previously proven high alkaloid, flavonoids, phenol, tannins content and free radical scavenging activity.

Phytochemical analysis and hypotensive potential of Teucrium stocksianum Boiss.

Teucrium stocksianum Boiss. is an aromatic perennial herb. It has long been used traditionally in the treatment of hypertension in northern areas of Pakistan. The aim of this study was to evaluate its folkloric claim as hypotensive plant, phytochemical analysis and to predict potential phytoconstituent through in-silico studies. Hypotensive effect was investigated in anesthetized normotensive Sprague-Dawley rats. Recording of chronotropic and inotropic effect of plant extract in isolated right atria was done using tissue organ bath technique. Further, phytochemical characterization was performed through LC-MS. Whereas docking studies were carried out against M2 mAchR and Ca2+ Channel receptor. Dose dependent reduction in systolic, diastolic, mean arterial pressure and heart rate was observed. Pretreatment with atropine and amlodipine significantly (p<0.001) reduced the hypotensive and negative chronotropic and inotropic effect. Phytochemical studies revealed the presence of twenty active compounds including Luteolin, Sarmentosin epoxide and Quinic acid. Docking studies showed pronounced interactions of majority of these phytochemicals with M2 mAch receptor in agonistic way and Ca2+ Channel receptor in antagonistic way. Results speculate that dose dependent hypotensive and bradycardia effect of Teucrium stocksianum are mediated through muscarinic pathway and Ca2+antagonism and is also well predicted by in-silico studies.

Phytochemical analysis and reappraisal of diuretic activity of Delphinium brunonianum Royle and its mode of action in experimental rats.

The aim of this study was the evaluation of diuretic potential of Delphinium brunonianum. Acute diuretic effect in rats was evaluated 8 h after administration of various doses of crude extract, fractions and hydrochlorthiazide. While, prolonged effect of butanolic fraction was assessed after 7days of oral administration in rats. Thereafter, involvement of different pathways in diuretic activity was also appraised. Furthermore, polyphenolic contents in butanolic fraction were assessed using HPLC/UV-VIS technique. All doses of extract and fractions induced a prominent increase in urine and Na+ excretion with no effect on excretion of K+. Prior administration of indomethacin and atropine considerably avoided the diuretic effect of butanolic fraction. Regarding the quantitative chemical analysis the polyphenolic contents were recorded as 28.78 µg/mg. Thus results of present investigation suggested that Delphinium brunonianum possess remarkable diuretic potential.

Study on vascular mechanisms underlying the hypotensive effect of Sorghum halepense (L.) Pers.

Sorghum halepense L (Poaceae), ordinarily it is known as Johnson grass and locally as baru. This study was designed to find the vascular mechanisms underlying the hypotensive activity of S. halepense. In this study, effect of S. halepense seed extract/fractions on various blood pressure parameters were evaluated in normal and fructose induced hypertensive rats by invasive technique. Possible underlying hypotensive mechanism of active fraction was determined by using various pharmacological inhibitors. S. halepense extract/fractions vasorelaxant effect were also evaluated on rat aorta rings in organ bath and various intracellular signaling pathway inhibitors were used for determination of underlying mechanisms. S. halepense extract/fractions produced blood pressure lowering effect with most significant effect by its aqueous soluble fraction at dose of 10mg/kg. This effect was attenuated by pretreatment of atropine. Aqueous soluble fraction produced endothelium dependent vasorelaxation in rat aortic rings that was inhibited by pretreatment of atropine after phenylephrine induced contraction. The vasorelaxant effect of aqueous soluble fraction was attenuated by potassium channel blockers and also produced inhibitory effect on calcium entry through calcium channels. It also suppressed phenylephrine induced contraction like verapamil. By HPLC analysis found vanillic acid and naringinin in it. In conclusion, aqueous soluble fraction of S.halepense possess phytoconstituents which may be responsible for hypotensive and vasorelaxant effect of Sorghum halepense.

Blood pressure lowering effects of Ranunculus scleratus Linn. in normal and fructose induced hypertensive rats and estimation of underlying mechanisms.

Ranunculus scleratus Linn. is used in folk medicine to treat hypertension. This study was aimed at providing validation to its traditional use and to explore underlying mechanisms of action. Effects of hydro-ethanolic crude extract of the plant and its fractions on blood pressure was evaluated using direct surgical method in normotensive and in fructose induced hypertensive rats. Various doses of crude extract, RSC, (5, 10, 20, 30mg/kg) and all fractions (3, 5, 10, 20mg/kg) were studied. Results suggested that aqueous fraction of R. scleratus (RSA) produced most pronounced effects at 10mg/kg in normotensive and at 20mg/kg in hypertensive animals. Underlying mechanisms, using various pharmacological antagonists were also elucidated. Results suggested the involvement of muscarinic receptor, angiotensin converting enzyme (ACE) inhibition, ganglionic block and nitric oxide (NO) release in presenting hypotensive response.

Evaluation of effect of Acacia jacquemontii Benth. on blood pressure in normotensive and fructose induced hypertensive sprague dawley rats: An ethnopharmacological approach.

Acacia jacquemontii Benth. is used traditionally to treat hypertension but no scientific literature supports this claim. So, this study was aimed at validating this claim. This was done by injecting various doses of crude extract of Acacia jacquemontii, AJC (5, 10, 20, 30mg/kg) and all fractions (hexane, ethyl acetate, n-butanol and aqueous) (3, 5, 10, 20mg/kg) intravenously in anaesthetized rat. Based on the results, butanol fraction (AJB) at 20mg/kg was found to be the most potent, so it was selected for exploring mechanisms of action. For this purpose, different groups were injected with various pharmacological inhibitors (L-NAME, atropine, captopril, propranolol and hexamethonium) prior to AJB administration. Also, AJB at 20mg/kg was evaluated for prolonged hypotensive effect for the period of 40 min. Results showed a significant dose dependent reduction in BP in normotensive and in hypertensive rats. AJC and AJB produced a decline in SBP, DBP and MAP with p<0.05 - p<0.001 and p<0.001 respectively in normotensive animals. Whereas in hypertensive animals, AJC showed significant reduction at 5mg/kg with p<0.01 and at 10, 20 and 30 mg/kg with p<0.001. AJB produced a decline in hypertensive animals at all tested doses with p<0.001. AJB resulted in hypotensive effect mediated by ß receptors, ganglionic block operating central sympathetic neural responses and renin angiotensin aldosterone system (RAAS). This study supports the ethnomedicinal claim of Acacia jacquemontii Benth. in treating hypertension.

Comparative hypoglycemic activity of different fractions of Thymus serpyllum L. in alloxan induced diabetic rabbits.

The aim of present study was to evaluate and compare the hypoglycemic activity of different solvents extracts of Thymus serpyllum in rabbits. Diabetes was induced with single intravenous injection of alloxan monohydrate (150mg/kg). Glibenclamide and acarbose were used as standard drugs. The crude powder of Thymus serpyllum (500 mg/kg b.w) significantly reduced blood glucose level in both normal and diabetic rabbits. Various extracts of Thymus serpyllum were compared for their hypoglycemic activity in diabetic rabbits. Ether and aqueous extracts significantly reduced the blood glucose level with maximum effect (p<0.001) produced by aqueous extract, which was selected for further study. Aqueous extract significantly inhibited the rise in glucose level in oral glucose tolerance test. The extract showed synergistic effect with different doses of insulin; however serum insulin level of the diabetic rabbits was not significantly increased by the extract. HbA1c level was significantly (p<0.05) reduced whereas hemoglobin level was significantly increased in three months study. Phytochemical screening of the aqueous extract showed the presence of alkaloids, flavonoids, tannins, terpinoids, reducing sugar and cardiac glycosides. It is concluded that the aqueous extract might be used alone or in combination with insulin to manage diabetes and its associated complications.