Nitin P. Kurade

A review of the toxicosis and biological properties of the genus Eupatorium

Eupatorium genus grows wild in many parts of the world. A number of species of Eupatorium are toxic to grazing animals. Milk sickness in humans is caused by ingestion of milk of the animals reared on the pastures infested with Eupatorium rugosum (white snakeroot). While some information is available on the toxins in various species of Eupatorium, ambiguities still persist in extrapolation of the data to field incidence of toxicosis. Eupatorium genus has been used for its medicinal properties for many decades. A number of bioactive natural products have been reported in the extracts of Eupatorium spp. and the genus is a promising bioresource for preparation of drugs and value-added products.

Chemical composition and antibacterial activity of essential oils of Lantana camara, Ageratum houstonianum and Eupatorium adenophorum.

Essential oils have applications in folk medicine, food preservation, and as feed additives. The essential oils of Lantana camara Linn. (Verbenaceae), Ageratum houstonianum Mill. (Asteraceae) and Eupatorium adenophorum Spreng. (Asteraceae) were analyzed by Gas chromatography-mass spectrometry (GCMS). In L. camara oil, of the total identified (83.91%) volatile constituents, five constituents [3,7,11-trimethyl-1,6,10-dodecatriene (28.86%), β-caryophyllene (12.28%), zingiberene (7.63%), γ-curcumene (7.50%) and α-humulene (3.99%)] represented the major ones. In A. houstonianum oil, among the total identified volatile constituents (94.51%), three [precocene-II (52.64%), precocene-I (22.45%) and β-caryophyllene (9.66%)] represented the major ones. In E. adenophorum oil, of the total identified volatile constituents (84.95%), six [1-napthalenol (17.50%), α-bisabolol (9.53%), bornyl acetate (8.98%), β-bisabolene (6.16%), germacrene-D (5.74%) and α- phellandrene (3.85%)] represented the major ones. The antibacterial activity expressed as Minimum Bactericidal Concentration (MBC) (μg/mL) was determined by the broth dilution method. The essential oil of E. adenophorum had antibacterial activity against Arthrobacter protophormiae, Escherichia coli, Micrococcus luteus, Rhodococcus rhodochrous, and Staphylococcus aureus with MBC values of 200, 100, 100, 12.5, and 200, respectively. The essential oil of A. houstonianum showed antibacterial activity against M. luteus and R. rhodochrous with MBC of 100 and 12.5, but not against A. protophormiae, E. coli, and S. aureus. The essential oil of L. camara showed antibacterial activity against A. protophormiae, M. luteus, R. rhodochrous and S. aureus with MBC of 50, 25, 12.5, and 200, respectively, but not against E. coli. MBC was lowest for R. rhodochrous for all the three essential oils.

Topically applied substance P enhanced healing of open excision wound in rats.

Significant social and financial burden due to wounds need newer drugs/formulations to speed up the healing process. Substance P (SP), a neuropeptide, is associated with release of various cytokines and growth factors from inflammatory, epithelial and endothelial cells. In the present study, temporal effects of topically applied SP (10(-7)M in normal saline) were evaluated in the modulation of various cytokines and growth factors that participate in cutaneous wound healing. Gross examination of full thickness open excision wound in rats revealed that once daily topical application of SP significantly increased the wound closure, as compared to control group. SP treatment significantly increased tumor necrosis factor-α (TNF-α) and decreased interleukin 10 (IL-10) levels on day 3. On the contrary, on day 7 level of TNF-α decreased and that of IL-10 increased. The mRNA and protein expressions of vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1) increased on days 3 and 7, and decreased on day 14 in SP-treated wounds. Histopathological evaluation of hematoxylin and eosin stained wound sections showed that SP treatment produced increased early leukocytes infiltration, fibroblast proliferation, angiogenesis, collagen deposition and re-epithelialization. Results of the present study demonstrate that topical application of SP enhanced wound healing by modulating cytokines, growth factors and cells. Based on the results, it is suggested that SP could be of beneficial use in diabetic wounds where levels of VEGF, TGF-β1 and SP decrease along with impairment of inflammatory reaction.

Topical pluronic F-127 gel application enhances cutaneous wound healing in rats.

Pluronic F-127 gel is used as vehicle for various topical applications. In the present study, effects of topical application of pluronic F-127 gel were evaluated in cutaneous wound healing in Wistar rats. Normal saline solution and pluronic F-127 gel (25%) were applied topically on open excision wounds for 14 days. Photography, determination of percentage wound contraction, and collection of granulation tissue were done on days 3, 7, 11 and 14 post-wounding. Topical application of gel (once daily) significantly increased the wound closure on days 11 and 14. The gel application increased the expressions of vascular endothelial growth factor (VEGF) and transforming growth factor-beta1 (TGF-β₁) on days 3 and 7. Histopathologically, more leukocyte infiltration followed by well formed granulation tissue with marked fibroblast proliferation was evident in the gel-treated group, as compared to the saline-treated control group. Immunohistochemistry of CD31 on day 7 revealed significant higher microvessel density in gel-treated wounds. Picrosirius staining demonstrated higher collagen fraction in gel-treated wounds. Thus, from the results, it could be concluded that pluronic F-127 gel has a mild inflammatory nature and enhanced the healing by stimulating expression of VEGF and TGF-β₁.

Hepatotoxicity of Eupatorium adenophorum to rats.

Freeze dried Eupatorium adenophorum leaf powder mixed in rat feed at a level of 25% elicited hepatotoxicity. The affected animals were jaundiced and had marked increase in plasma bilirubin levels and activities of alkaline phosphatase, glutamate oxaloacetate transaminase and glutamate pyruvate transaminase. The liver of intoxicated animals had focal areas of necrosis and bile duct proliferation. Elevation in plasma bilirubin concomitant with alterations in enzyme profile and histopathological lesions are consistent with liver injury and cholestasis. This is the first report of the toxicity of E. adenophorum to rats.

Atorvastatin restores arsenic-induced vascular dysfunction in rats: Modulation of nitric oxide signaling and inflammatory mediators

We evaluated whether atorvastatin, an extensively prescribed statin for reducing the risks of cardiovascular diseases, can reduce the risk of arsenic-induced vascular dysfunction and inflammation in rats and whether the modulation could be linked to improvement in vascular NO signaling. Rats were exposed to sodium arsenite (100ppm) through drinking water for 90 consecutive days. Atorvastatin (10mg/kg bw, orally) was administered once daily during the last 30days of arsenic exposure. On the 91(st) day, blood was collected for measuring serum C-reactive protein. Thoracic aorta was isolated for assessing reactivity to phenylephrine, sodium nitroprusside and acetylcholine; evaluating eNOS and iNOS mRNA expression and measuring NO production, while abdominal aorta was used for ELISA of cytokines, chemokine and vascular cell adhesion molecules. Histopathology was done in aortic arches. Arsenic did not alter phenylephrine-elicited contraction. Atorvastatin inhibited Emax of phenylephrine, but it augmented the contractile response in aortic rings from arsenic-exposed animals. Sodium nitroprusside-induced relaxation was not altered with any treatment. However, arsenic reduced acetylcholine-induced relaxation and affected aortic eNOS at the levels of mRNA expression, protein concentration, phosphorylation and NO production. Further, it increased aortic iNOS mRNA expression, iNOS-derived NO synthesis, production of pro-inflammatory mediators (IL-1β, IL-6, MCP-1, VCAM, sICAM) and serum C-reactive protein and aortic vasculopathic lesions. Atorvastatin attenuated these arsenic-mediated functional, biochemical and structural alterations. Results show that atorvastatin has the potential to ameliorate arsenic-induced vascular dysfunction and inflammation by restoring endothelial function with improvement in NO signaling and attenuating production of pro-inflammatory mediators and cell adhesion molecules.

Biochemical alterations in the blood plasma of rats associated with hepatotoxicity induced by Eupatorium adenophorum.

Eupatorium adenophorum (Crofton weed), a native of Central America, has appeared as a major weed in several areas in different parts of the world. Horses that eat this plant are poisoned on prolonged exposure. Toxicity due to consumption of this plant by other grazing animals is not clear. Administration of freeze-dried leaf powder to mice results in hepatotoxicity. Earlier attempts to produce toxicity in rats using the leaves of this plant were not successful. In the present study, administration of oven-dried E. adenophorum leaves collected at the flowering stage elicited hepatotoxicity in rats. The affected animals had a marked increase in the concentration of plasma bilirubin and in the activities of 5′-nucleotidase, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase. There were no significant differences in plasma creatinine, urea or total protein values in the affected animals compared to controls. The livers of the affected animals had focal areas of necrosis throughout the parenchyma and hepatocytes showed megalocytosis. The bile ducts were dilated and the epithelium showed degenerative to necrotic changes. The alterations in bilirubin, enzymes and histopathological changes imply cholestasis and liver injury.