Pharmacological basis for the antihypertensive activity of Grewia asiatica fruit extract.

In the management of cardiovascular disorders, medicines from herbal sources have played a vital role through centuries. The following study was commenced in order to lay possible pharmacological foundation associated with medicinal uses of edible fruit of Grewia asiatica in hypertension through in-vitro method. In this study isolated atrial preparation of Guinea pig was used where crude ethanolic extract of Grewia asiatica fruit (Ga.Cr) decreased the force and rate of spontaneous atrial contractions (0.03-10mg/kg). In isolated rat aortic ring preparations previously vasoconstricted by phenylephrine and High K+, it also resulted in dose dependent vasodilation (0.01-10 mg/kg).In the presence of L-NAME, the relaxation curve of Ga.Cr was partially inhibited showing involvement of Nitric oxide (NO) mediated pathway. The speculative analysis contemplated that Ga.Cr has blood pressure reducing potentials through inhibition of Ca++ influx via Ca++ channels, its release from intracellular stores and through other means like NO mediated pathways.

Pharmacologic targeting of Cdc42 GTPase by a small molecule Cdc42 activity-specific inhibitor prevents platelet activation and thrombosis.

Gene targeting of Cdc42 GTPase has been shown to inhibit platelet activation. In this study, we investigated a hypothesis that inhibition of Cdc42 activity by CASIN, a small molecule Cdc42 Activity-Specific INhibitor, may down regulate platelet activation and thrombus formation. We investigated the effects of CASIN on platelet activation in vitro and thrombosis in vivo. In human platelets, CASIN, but not its inactive analog Pirl7, blocked collagen induced activation of Cdc42 and inhibited phosphorylation of its downstream effector, PAK1/2. Moreover, addition of CASIN to washed human platelets inhibited platelet spreading on immobilized fibrinogen. Treatment of human platelets with CASIN inhibited collagen or thrombin induced: (a) ATP secretion and platelet aggregation; and (b) phosphorylation of Akt, ERK and p38-MAPK. Pre-incubation of platelets with Pirl7, an inactive analog of CASIN, failed to inhibit collagen induced aggregation. Washing of human platelets after incubation with CASIN eliminated its inhibitory effect on collagen induced aggregation. Intraperitoneal administration of CASIN to wild type mice inhibited ex vivo aggregation induced by collagen but did not affect the murine tail bleeding times. CASIN administration, prior to laser-induced injury in murine cremaster muscle arterioles, resulted in formation of smaller and unstable thrombi compared to control mice without CASIN treatment. These data suggest that pharmacologic targeting of Cdc42 by specific and reversible inhibitors may lead to the discovery of novel antithrombotic agents.

Efficiency of proteolytic enzymes in treating lumbar spine osteoarthritis (low back pain) patients and its effects on liver and kidney enzymes.

Lumbar spine osteoarthritis with 40-85% prevalence, degeneration of spine with remarkable narrowing of disc space and osteophytes formation trigger pain in lower back. Pain in lower portion of back is now considered to be the second most commonly treated health issue in primary health care setups. This pain causes disability, functional loss and job absentees. Commonly pain is managed pharmacologically by NSAIDS but resulted in severe gastric side effects. The purpose of this trial was to appraise the properties of bromelain and papain, the vegetal proteolytic enzymes, in comparison with standard drug on LBP patients. Forty men and women with lumbar spine osteoarthritis were recruited and divided into group 1, received aceclofenac 100mg tablet b.i.d as standard treatment, group 2, patients treated with aceclofenac 100 mg tablet b.i.d and enzyme supplements 250 mg b.i.d for 6 weeks. All the participants were evaluated for their body mass index, vital signs and liver/kidney enzymes before and after treatment. Moreover intensity of pain were also measured through visual analogue scale (VAS) and oswestry low back pain questionnaire (ODI) before treatment (0 week), 3rd week and 6th week of treatment. The enzyme group patients showed significantly diminished pain scores VAS from 7.10±1.29 to 5.85±1.531*** (P=0.001), ODI score from 56.2±8.70 to 51.6±8.125*** (P=0.000), significantly diminished enzymes; ALP from 210.00±55.24 to 196.90±51.02 (P=0.054*) and serum creatinine from 0.97±0.153 to 0.87±0.139 (P=0.035*) and improved quality of life. Hence, this study suggested that the enzyme supplements for 6 weeks have prolonged beneficial carry-over effects in comparison to standard treatment without producing any change in BMI (P>0.05) and vital signs (P> 0.05).

Histopathological effects on stomach, liver and kidney associated with Cedrus deodara root oil in ulcer induced rats (Wistar strain).

Cedrus deodara have been used traditionally in ayurvedic system against peptic ulcer. Present work is concerned with the determination of histopathological effects in ethanol induced ulcer on rats (Wistar Strain) treated with Cedrus deodara root oil at a dose of 200mg/kg and comparison of its antiulcer activity against control, positive control and standard anti-ulcer drug (Omeprazole). The aim was to find out the antiulcer effect of Cedrus deodara root oil and to observe histopathology of liver, kidney as well. 120 Albino rats were taken and divided into four groups i.e. A, B, C and D designated as control, positive control, standard and treated groups respectively. Normal and intact general architecture of mucosa and submucosa layers of stomach observed. No significant changes observed in thickness of epithelium, no inflammatory cells were present on the mucosa and submucosal layer and gastric glands were normal. Liver of albino rats, showed no dilation and congestion in central as well as portal vein. Kidney of albino rats exhibited no shrinkage in glomeruli, no congested and dilated renal corpuscles, neither hemolysis nor congested and dilated renal tubules were seen. It is concluded that C. deodara root oil has anti-ulcer properties without effecting kidney and liver tissues.

Effects of aqueous leaves extract of Holoptelea integrifolia (Roxb) Planch on liver and kidney histopathology of albino rats.

Histopathological studies are an essential element to ascertain comprehensive safety profile of a drug. Unfortunately limited data are available about the toxicity of herbal remedies. Since a popular medicinal plant Holoptelea integrifolia (Roxb) Planch. contains various bioactive molecules, the present study is aimed to assess the histopathological alterations induced by aqueous extract of Holoptelea integrifolia on liver and kidney of wistar albino rat. In this study 60 rats divided in two groups; control and treated with aqueous extract of Holoptelea integrifolia (250mg/kg body weight) for 5 days. Histopathlogical studies by hematoxylin and eosin (H&E) staining were done on the liver and kidney tissues at the end of dosing by using standard procedure. Microscopic examination was then carried out to observe any pathological changes in the animals. The result showed that there is no significant variation in the basic architecture of liver and kidney as compared to control male wistar albino rats. In conclusion, aqueous extract of leaves of H. integrifolia may be safe and nontoxic. Further work on pharmacological aspects is required to evaluate the clinical potential of this plant for different ailments.

Hepatoprotective and antidiabetic effect of Guaiacum officinale in diabetes induced male albino wistar rats.

The aim of present study is to evaluate the antidiabetic and hepatoprotective effect of Guaiacum officinale in streptozotocin induced male albino rats. The methanolic bark extract of Guaiacum officinale was administered at a dose of 500 mg/kg and Glibenclamide was used as a standard drug at a dose of 0.5mg/kg for 28 days. The animals were divided in to four groups. Control group n=12, received distilled water. Positive control group (STZ group) n=12 received streptozotocin at 30 mg/kg dose through I/P route. Standard group (STZ+ GLB group): n=12 received Glibenclamide. Treated group (STZ+ extracted group): n=12 received bark extract of Guaiacum officinale. Blood glucose level was significantly reduced after oral adminstration of bark extract in streptozotocin induced diabetic rats. The SGOT level significantly reduced in Guaiacum officinale treated group as compare to control, pronounced reduction of ALT level as compared to GLB and the ALP levels was highly significantly reduced in Guaiacum officinale treated group while GLB is unable to improve ALP levels in GLB treated diabetic albino rats. The level of direct bilirubin in Guaiacum officinale treated group was found to be insignificant as compared to control and STZ treated group while the level of indirect bilirubin was significantly reduced in STZ treated group as compare to control. Histopathological studies showed that Guaiacum officianle have hepatoprotective effect in experimental induced male albino rats.

Report: effects of Camellia sinensis L. (green tea) extract on the body and testicular weight changes in adult Wistar rate.

This research was aimed to study the effects of oral administration of Camellia sinensis L. on the testicular and body weights of adult Wistar rats for short and long time periods. The adult Wistar rats were divided into 3 groups (A, B and C). Every group had ten rats. Green tea extract 0.692% (w/v) was given to groups A and B on daily basis. The extracts were prepared fresh and given for a period of ten and thirty days, respectively, while distilled water was given to the group C rats only. The adult Wistar rats were sacrificed on eleventh and thirty-first day of experiment for the particular groups. The testes were dissected out cautiously, free from the supporter tissues and weighed to the adjacent 1 mg. There is no significant difference in the body weight in all 3 groups. Moreover, it was observed that Wistar rat's testicular weight was considerably increased in group B but no major changes were seen in group A. Our results indicated that green tea when given for short period of time may be effective to the testes but has no consequence on Wistar rat's body weight. However, it is indistinct if these alterations are reversible.

Histopathologic changes in liver and kidney of male sprague dawley rats treated with extract of Cardiospermum halicacabum L.

In traditional medicine Cardiospermum halicacabum L. (Sapindeaceae) is used against various ailments such as rheumatism, nervous diseases, stiffness of the limbs and snakebite. Leaves are crushed and made into a tea, which aids itchy skin. Salted leaves are used as a poultice on swellings. Young leaves can be cooked and used as vegetables. The leaf juice has been used as a treatment for earache as well. In this study we evaluate acute toxicity (10, 50, 100 and 500 mg/kg) and pathologic changes in esophagus, stomach, liver and kidney tissues with a magnifying glass and microscope in a row to mark changes to both morphological and histological in comparison to control with the treatment of ethyl acetate extract (dose of 40mg/kg) in male Sprague Dawley rats. The rats were divided into 4 groups consisting of 3 rats per group for acute toxicity and histopathological change. In conclusion, no lethality was observed in acute toxicity study for 7 days. The treatment of ethyl acetate extracts at 40 mg/kg did not show lethal toxicological changes as observed by histopathological examination in the kidney and liver tissues.

Assessment of Cedrus deodara root oil on the histopathological changes in the gastrointestinal tissues in rats.

The present study was designed to investigate the effect of Cedrus deodara root oil on the histopathology of different gastrointestinal organs of Wistar rats. This oil was used traditionally as an anti-ulcer agent in the Indus Unic System and extracted from the plant root by destructive distillation method. A total of 90 rats were taken and divided into groups A, B and C, each comprising of 30 animals. The animals of group B and C were given 0.5 ml/kg and 2.5 ml/kg of C. deodara oil respectively while group A served as control and administered vehicle only. The treatment was given to the animals ones only for 24 hours. All animals were sacrificed and the organs like esophagus, stomach and ileum were taken out. Tissue processing and staining procedure was then carried out for any pathological changes in the animal tissues during microscopic examination. The results indicated that Cedurs deodara root oil at both doses 0.5ml/ kg and 2.5 ml/kg exhibited some adverse effects such as erosion of epithelium, edema on sub-mucosal and mucosal layers, congestion of blood vessels as well as presence of inflammatory cells on esophagus, stomach and ileum were seen. Moreover shortening of villi was also seen at both doses. A study conducted on mammalian toxicity previously on rats revealed that the C. deodara root oil used is not very toxic and comes under least toxic group as standardized by toxicologists. Based on the results obtained it was concluded that C. deodara root oil produced some adverse changes in the tissues of GIT when given at 0.5 ml/kg and 2.5 ml/kg doses but the effects were not lethal therapeutically at this dose LC50 16.5 ml/kg. The plant oil showed some toxicity and needs further detailed studies to assess its potential toxicity and therapeutic status before using this material as drug.

Analgesic activity of various extracts of Holoptelea integrifolia (Roxb.) Planch leaves.

The various extracts of leaves Holoptelea integrifolia (Ulmaceae) were investigated for analgesic activity in mice by tail flick method. The fresh plant leaves of H. integrifolia were collected, dried, cleaned, weighed and chopped into small pieces and percolated in ethanol. The fractionation of crude extract, followed by the addition of distilled water, ethyl acetate and n-butanol to an aqueous portion of each solvent, to obtain the dried masses of each four layers. Qualitative chemical examination indicates the presence of secondary metabolites such as alkaloids, flavones, phenol, steroids, tannins and triterpenoids. No acute oral toxicity was observed and extracts considered being safe at the dose of 50-2000 mg/kg body weight. At the dose of 500 mg/kg various extracts of leaves of H. integrifolia were found statistically significant (P<0.05). A maximum effect was established at 150 min, after drug administration. Diclofenace sodium used as a standard.

A novel small molecule 1,2,3,4,6-penta-O-galloyl-α-D-glucopyranose mimics the antiplatelet actions of insulin.

BACKGROUND: We have shown that 1,2,3,4,6-penta-O-galloyl-α-D-glucopyranose (α-PGG), an orally effective hypoglycemic small molecule, binds to insulin receptors and activates insulin-mediated glucose transport. Insulin has been shown to bind to its receptors on platelets and inhibit platelet activation. In this study we tested our hypothesis that if insulin possesses anti-platelet properties then insulin mimetic small molecules should mimic antiplatelet actions of insulin. PRINCIPAL FINDINGS: Incubation of human platelets with insulin or α-PGG induced phosphorylation of insulin receptors and IRS-1 and blocked ADP or collagen induced aggregation. Pre-treatment of platelets with α-PGG inhibited thrombin-induced release of P-selectin, secretion of ATP and aggregation. Addition of ADP or thrombin to platelets significantly decreased the basal cyclic AMP levels. Pre-incubation of platelets with α-PGG blocked ADP or thrombin induced decrease in platelet cyclic AMP levels but did not alter the basal or PGE(1) induced increase in cAMP levels. Addition of α-PGG to platelets blocked agonist induced rise in platelet cytosolic calcium and phosphorylation of Akt. Administration of α-PGG (20 mg kg(-1)) to wild type mice blocked ex vivo platelet aggregation induced by ADP or collagen. CONCLUSIONS: These data suggest that α-PGG inhibits platelet activation, at least in part, by inducing phosphorylation of insulin receptors leading to inhibition of agonist induced: (a) decrease in cyclic AMP; (b) rise in cytosolic calcium; and (c) phosphorylation of Akt. These findings taken together with our earlier reports that α-PGG mimics insulin signaling suggest that inhibition of platelet activation by α-PGG mimics antiplatelet actions of insulin.

Gene targeting implicates Cdc42 GTPase in GPVI and non-GPVI mediated platelet filopodia formation, secretion and aggregation.

BACKGROUND: Cdc42 and Rac1, members of the Rho family of small GTPases, play critical roles in actin cytoskeleton regulation. We have shown previously that Rac1 is involved in regulation of platelet secretion and aggregation. However, the role of Cdc42 in platelet activation remains controversial. This study was undertaken to better understand the role of Cdc42 in platelet activation. METHODOLOGY/PRINCIPAL FINDINGS: We utilized the Mx-cre;Cdc42(lox/lox) inducible mice with transient Cdc42 deletion to investigate the involvement of Cdc42 in platelet function. The Cdc42-deficient mice exhibited a significantly reduced platelet count than the matching Cdc42(+/+) mice. Platelets isolated from Cdc42(-/-), as compared to Cdc42(+/+), mice exhibited (a) diminished phosphorylation of PAK1/2, an effector molecule of Cdc42, (b) inhibition of filopodia formation on immobilized CRP or fibrinogen, (c) inhibition of CRP- or thrombin-induced secretion of ATP and release of P-selectin, (d) inhibition of CRP, collagen or thrombin induced platelet aggregation, and (e) minimal phosphorylation of Akt upon stimulation with CRP or thrombin. The bleeding times were significantly prolonged in Cdc42(-/-) mice compared with Cdc42(+/+) mice. CONCLUSION/SIGNIFICANCE: Our data demonstrate that Cdc42 is required for platelet filopodia formation, secretion and aggregation and therefore plays a critical role in platelet mediated hemostasis and thrombosis.