Protection against osteoarthritis in experimental animals by nanogold conjugated snake venom protein toxin gold nanoparticle-Naja kaouthia cytotoxin 1.

BACKGROUND & OBJECTIVES: Increased severity of osteoarthritis (OA) and adverse side effects of its treatment led to the search for alternative therapies. It was previously reported that snake venom protein toxin Naja kaouthia cytotoxin 1 (NKCT1) and gold nanoparticle (GNP) individually have potential against excremental arthritis. In this study, we analyzed the protective activity of GNP conjugated protein toxin NKCT1 (GNP-NKCT1) against experimental OA. METHODS: Gold nanoparticle conjugation with NKCT1 (GNP-NKCT1) was done and its physiochemical properties were studied. OA was induced in male albino rats by intra-articular injection of bacterial collagenase and treatment was done with NKCT1/GNP-NKCT1/standard drug (indomethacin). Physical parameter (ankle diameter), urinary markers (hydroxyproline, glucosamine, pyridinoline, deoxypyridinoline), serum and synovial membrane pro-inflammatory markers [tumour necrosis factor-alpha (TNF-α), interleukin-1ß (IL-1ß), IL-17, vascular endothelial growth factor (VEGF)] and matrix metalloproteinase 1 (MMP1) were measured. Joint histopathology and scanning electron microscopy imaging of articular cartilage surface were also done. RESULTS: Physical parameters, urinary markers, serum and synovial membrane pro-inflammatory makers and MMP1 were increased in arthritic rats and significantly restored after GNP-NKCT1/NKCT1 treatment. Joint histopathology and scanning electron microscopy imaging of articular cartilage surface also indicated the protective effect of GNP-NKCT1 against inflammatory response and cartilage degradation in osteoarthritic rats. INTERPRETATION & CONCLUSIONS: In this study restoration of the arthritic markers and bone degradation by GNP-NKCT1 treatment indicated the anti-osteoarthritic property of GNP-NKCT1. Further studies need to be done to confirm these findings.

Effect of Naja naja Laurenti shed skin extract on estrous cycle, hormone - cytokine profiles, histopathology of ovary and uterus of Swiss albino mice.

The snake shed skin though considered as biological waste products have been mentioned in folk and traditional medicine for treatment of ailments like skin disorders, parturition problems etc. Shedded skin extract (5 mg.kg-1, sc) did not produce any change in the estrous cycle of normal cycling female mice. However in 10 mg.kg-1, sc dose, the extract caused a temporary cessation of the estrous cycle at diestrous phase in normal cycling female mice for 10 days. SSAE (10 mg.kg-1, sc) caused a significant change in the level of LH, FSH, progesterone, estradiol, IL-beta, IL-6 and TNF-alpha. Histopathology of uterus and ovary showed structural disorientation in both. The results substantiate the influence of snake shed skin in mice reproductive cycle.

Ethno biological usage of zoo products in rheumatoid arthritis.

Rheumatoid arthritis (RA) is one of the most common autoimmune disorder which causes swelling, redness, pain, stiffness, restriction of limb movements, decreases life expectancy and early death of the patients. Available drugs include non steroidal anti-inflammatory and analgesics, disease modifying anti-rheumatic drugs and steroids (glucocorticoids etc). All these drugs have their own limitations such as gastrointestinal irritations, cardiovascular problems, and drug dependency. Search for alternative therapy from natural products are being ventured throughout the world. Zoo therapy in arthritis, a common practice of the ancient times that have been mentioned in traditional and folk medicine. The scientific basis of some of the zoo products are being explored and have been showing promising results in experimental rheumatoid arthritis. These therapies have minimum side effects and many of them have potential to give rise to drug development clues against rheumatoid arthritis. The present review is an effort to establish the folk and traditional treatment of rheumatoid arthritis using zoo products.

Anticancer potential of animal venoms and toxins.

Anticancer drug development from natural resources are ventured throughout the world. Animal venoms and toxins a potential bio resource and a therapeutic tool were known to man for centuries through folk and traditional knowledge. The biodiversity of venoms and toxins made it a unique source of leads and structural templates from which new therapeutic agents may be developed. Venoms of several animal species (snake, scorpion, toad, frog etc) and their active components (protein and non protein toxins, peptides, enzymes, etc) have shown therapeutic potential against cancer. In the present review, the anticancer potential of venoms and toxins from snakes, scorpions, toads and frogs has been discussed. Some of these molecules are in the clinical trials and may find their way towards anticancer drug development in the near future. The implications of combination therapy of natural products in cancer have been discussed.

Gold Nanoparticles (AuNPs) Conjugated with Andrographolide Ameliorated Viper (Daboia russellii russellii) Venom-Induced Toxicities in Animal Model.

Andrographolide, a diterpenoid compound found in the aerial parts of Andrographis paniculata (a well known anti snake venom plant) was conjugated with gold nanoparticle (andrographolide-AuNPs) and its efficacy against Daboia russellii russellii venom (DRRV) induced local damage, organ toxicity and inflammatory response was evaluated in animal models. Ethical clearance was obtained before animal experiments. Andrographolide-AuNPs was formed by adsorption method. Physico-chemical characterization of particle was done by dynamic light scattering (DLS), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM) and X-ray diffraction (XRD). Swiss albino male mice were divided into 5 groups: Gr. 1-Sham control, Gr. 2-DRRV control, Gr. 3-anti snake venom serum treated, Gr. 4-andrographolide treated and Gr. 4-andrographolide-AuNPs treated. 1/5th minimum lethal dose of DRRV (10 µg/s.c./20 g mice) was induced in animals of group 2, 3, 4 and 5 animals, followed by treatment with anti snake venom serum (2 mg/20 g mice, i.v.) andrographolide (50 µg/20g mice, i.p.) and andrographolide-AuNPs (50 µg/20 g mice, i.v.) in group 3, 4 and 5 animals, respectively. Blood was collected after 18 h, serum was prepared and organ toxicity markers (transaminases, phosphatases, lactate dehydrogenase, creatine phosphate, urea, creatinine, Ca2+, phosphorous), inflammatory markers (interleukin 1ß, 6, 17a, 10, tumor necrosis factor α) and local damage testings (defibrination, edema, hemorrhage) were assessed. Values were expressed as mean ± SEM (n = 4), one way analysis of variance was done, P < 0.05 was considered as statistically significant. Formed andrographolide-AuNPs were pink in color with hydrodynamic diameter 30-50 nm, polydispersity index 0.412 and zeta potential -16.21 mV. XRD data confirmed the presence of crystalline gold in andrographolide-AuNPs. TEM (20-50 nm) and FE-SEM (20-25 nm) indicated the presence of nearly spherical particle. DRRV envenomation followed by treatment with andrographolide-AuNPs provided protection against venom induced edema, hemorrhage, defibrination, organ toxicity and inflammation in animal model. Venom neutralization by andrographolide-AuNPs was > andrographolide, which confirmed the increased efficacy of andrographolide after gold nanoparticle conjugation, may be due to anti-oxidant/anti-inflammatory activity of andrographolide, showing increased efficacy after gold nanoparticle tagging. Thus, andrographolide-AuNPs may serve as a supportive therapy in snakebite (against venom induced local damage, organ toxicity and inflammatory response) subject to further detail studies.

Anti-fibrotic effect of black tea (Camellia sinensis) extract in experimental pulmonary fibrosis.

There is no effective therapy exists for Idiopathic pulmonary fibrosis (IPF) till now. Few studies have been done on protective effects of green tea in pulmonary fibrosis but there is no single report on black tea extract (BTE) in pulmonary fibrosis so far. This study aims to investigate the anti-fibrotic effect of BTE against experimental pulmonary fibrosis. Four groups of animals were selected for this study. Group 1: control group mice. Group 2: mice exposed to bleomycin for 21 days, Group 3 and Group 4: bleomycin exposed mice treated with 25 mg BTE/kg b.w./day, p.o and 50 mg BTE/kg b.w./day, p.o. respectively for 21 days. Bleomycin exposed mice showed increased collagen deposition and wet/dry weight ratio, which were attenuated upon 50 mg BTE/kg b.w. treatment. The increased level of histopathological parameters in bleomycin-induced mice was significantly decreased after 50 mg BTE/kg b.w. treatment. Furthermore, 50 mg BTE/kg b.w. administration also decreased the expression of α-SMA in bleomycin-induced mice. This treatment with 50 mg BTE/kg b.w. also down regulated the expression of TGF-ß and up regulated IFN-γ expression in experimental pulmonary fibrosis. The results of the present study put-forward BTE as a potential anti-fibrotic agent due to its attenuating effect on potential fibrotic markers.

Black Tea (Camellia sinensis) Extract Induced Prenatal and Postnatal Toxicity in Experimental Albino rats.

BACKGROUND: Tea (Camellia sinensis) being the most widely drank beverage and despite having numerous beneficial role toward health and disease, its safety evaluation during pregnancy and prenatal, postnatal developmental period need to be monitored. OBJECTIVE: This study was to evaluate the toxicity of black tea extract (BTE) in experimental pregnant rats and on their pups during prenatal and postnatal developmental periods. MATERIALS AND METHODS: Pregnant female (120 ± 10 g) Wister albino rats were chosen for this study. Group 1 was control group where pregnant female rats were treated with saline. Group 2 and Group 3 were pregnant female rats treated with 50 mg and 100 mg BTE/kg/day, respectively, throughout prenatal and postnatal periods. All three groups of rats were provided food and drinking water ad libitum. Animals were examined through their urinary and serum parameters, histopathological studies, and biomorphometric studies in pups. All data were expressed as mean ± standard deviation with significance between the controls and the treated groups (n = 6). Collected data were subjected to the analysis of variance and Tukey test; P < 0.05 was considered as statistically significant. RESULTS: BTE produced significant alterations in urinary calcium, creatinine, and urea during prenatal period; exhibited proteinuria, ketonuria, and histology showed nephrotoxicity during postnatal period, and BTE also showed a significant increase in serum proinflammatory cytokines and decreased anti-inflammatory cytokines level compared to control group. BTE caused significant changes in biomorphometric parameters in the pups as compared with pups of control mothers. CONCLUSION: This study confirmed the BTE-induced toxicity in pregnant rats and their pups. SUMMARY: Black tea (Camellia sinensis) is the most widely drank beverage. This study was to evaluate the toxicity BTE in experimental pregnant rats and on their pups during prenatal and postnatal developmental periods. Animals were examined through their urinary and serum parameters, histopathological studies, and biomorphometric studies in pups. BTE.induced toxicity in pregnant rats and their pups. Abbreviations used: BTE: Black tea extract, IL-1α: Interleukin 1 alpha, IL-1 ß: Interleukin 1 beta, IL-6: Interleukin 6, IL-10: Interleukin 10, TNF-α: Tumor necrosis factor alpha.

Anti-osteoarthritic activity of Bungarus fasciatus venom fraction BF-F47 involving molecular markers in the rats.

A heat stable protein BF-F47 was purified from the crude venom of Bungarus fasciatus by CM cellulose ion exchange chromatography and HPLC. Osteoarthritis (OA) was developed in male albino Wistar rats by collagenase injection. BF-F47 treatment significantly restored urinary hydroxyproline and glucosamine in OA rats. Serum acid phosphatase, alkaline phosphatase, creatinine and serum molecular markers TNF-α, IL-1ß, IL-17, cytokine induced neutrophil chemoattractant-1, matrix metalloproteinase-1, cathepsin-K, osteocalcin and PGE2 were also significantly altered. BF-F47 showed partial restoration of osteoarthritis joints. Thus, BF-F47 induced anti-osteoarthritic activity in Wistar rats acted through molecular markers of arthritis and inflammation.

A high molecular weight protein Bengalin from the Indian black scorpion (Heterometrus bengalensis C.L. Koch) venom having antiosteoporosis activity in female albino rats.

This study reports the presence of a high molecular weight protein (Bengalin) from the Indian black scorpion (Heterometrus bengalensis) venom having antiosteoporosis activity in experimental osteoporosis developed in female albino Wister rats. Bengalin was purified through DEAE-cellulose ion exchange chromatography and high performance liquid chromatography. The molecular weight of the Bengalin was found to be 72kDa and the first 20 amino acid sequence was found to be G-P-L-T-I-L-H-I-N-D-V-H-A-A/R-F-E-Q/G-F/G-N-T. Bengalin exhibited significant antiosteoporosis activity in experimental female rats, which was confirmed through analysis of urine Ca(2+), PO(4)(3-), CRE & OH-P. Bengalin (3 microg and 5 microg/100g rat/i.p.) antagonized osteoporosis by restoring urinary Ca(2+), PO(4)(3-), CRE and OH-P, serum/plasma Ca(2+), PO(4)(3-), ALP, TRAP, PTH, T(3), TSH, Osteocalcin, IL1, IL6 and TNF alpha and bone minerals Ca(2+), P, Mg(2+), Zn(2+), Na(+), as compared with the sham operated control rats. Bone minerals density of osteoporosis female rats was improved due to Bengalin, observed through DEXA scan. Subacute toxicity studies in male albino mice, Bengalin showed cardiotoxicity. In vivo experiments, Bengalin showed cardiotoxicity on isolated guinea pig heart, guinea pig auricle, and neurotoxicity on isolated rat phrenic nerve diaphragm preparation. Further detail studies on the toxicity, antiosteoporosis and structural identity of Bengalin are warranted.