Earthworm extract attenuates silica-induced pulmonary fibrosis through Nrf2-dependent mechanisms.

Silicosis is an occupational pulmonary fibrosis caused by inhalation of silica (SiO2) and there are no ideal drugs to treat this disease. Earthworm extract (EE), a natural nutrient, has been reported to have anti-inflammatory, antioxidant, and anti-apoptosis effects. The purpose of the current study was to test the protective effects of EE against SiO2-induced pulmonary fibrosis and to explore the underlying mechanisms using both in vivo and in vitro models. We found that treatment with EE significantly reduced lung inflammation and fibrosis and improved lung structure and function in SiO2-instilled mice. Further mechanistic investigations revealed that EE administration markedly inhibited SiO2-induced oxidative stress, mitochondrial apoptotic pathway, and epithelial-mesenchymal transition in HBE and A549 cells. Furthermore, we demonstrate that Nrf2 activation partly mediates the interventional effects of EE against SiO2-induced pulmonary fibrosis. Our study has identified EE to be a potential anti-oxidative, anti-inflammatory, and anti-fibrotic drug for silicosis.

Efficacy of frog skin lipids in wound healing.

BACKGROUND: Frog skin has been sequentially and scientifically evaluated by our group for its wound healing efficiency. Owing to the complex structure of skin, attempts were being made to analyse the role of individual constituents in different phases of healing. Our earlier papers have shown the significance of frog skin not only in wound healing but also enhancing the proliferating activity of the epidermal and dermal cells which are instrumental for normal healing process. We also have identified for the first time novel antimicrobial peptides from the skin of Rana tigerina and thereby reduce the complications involved in the sepsis. PURPOSE OF THE STUDY AND RESULTS: The current study envisages the role of frog skin lipids in the inflammatory phase of wound healing. The lipid moiety of the frog skin dominated by phospholipids exhibited a dose dependent acceleration of healing irrespective of the mode of application. The efficiency of the extract is attributed partially to the anti-inflammatory activity as observed by the histochemical and immunostimulatory together with plethysmographic studies. CONCLUSIONS: Thus, frog skin for the first time has been demonstrated to possess lipid components with pharmaceutical and therapeutic potential. The identification and characterization of such natural healing molecules and evaluating their mechanism of action would therefore provide basis for understanding the cues of Nature and hence can be used for application in medicine.

[Study on the antitumor effects and synergism and attenuation effects of gecko alcohol extract].

OBJECTIVE: To investigate the antitumor effects of Gecko alcohol extract and its synergism and attenuation effects on CTX. METHOD: S180-bearing mice were given Gecko alcohol extract via intravenous injection,the tumor inhibitory rate and the levels of serum TNF-alpha of mice were detected. After inoculation of S180 tumor, the synergism and attenuation effects of Gecko alcohol extract on CTX were observed. After 12 days treatment, the tumor inhibitory rate, the count of peripheral white blood cells, index of thymus and spleen were calculated. RESULTS: Gecko alcohol extract in different dose (0.6, 1.2, 2.4 g/kg) inhibited the growth of S180 sarcoma in KM mice. The tumor inhibitory rates of 0.6, 1.2, 2.4 g/kg Gecko alcohol extract were 44.88%, 63.94%, 69.53%, respectively. However, the levels of serum TNF-alpha of mice not changed. The tumor inhibitory rates of intravenous administration of 0.6, 1.2, 2.4 g/ kg Gecko alcohol extract combined with CTX (20 mg/kg) were 56.93%, 67.15%, 70.24%, which were higher than that of CTX administration alone (41.71%). Compared with those in CTX group, the count of WBC (P < 0.01), the indexes of thymus and spleen (P < 0.05) were significantly elevated in all Gecko alcohol extract and CTX combination groups. CONCLUSION: Gecko alcohol extract has anti-tumor effects in vivo and attenuation effects on CTX.

[Isolation and pharmacological properties of analgesic fraction from venom of Naja Naja atra].

OBJECTIVE: To separate main analgesic fraction from venom of Guangdong Naja naja atra, to establish the basis for the using of Naja naja atra and find new analgesic fraction. METHODS: Affinity chromatography and size exclusion were used to isolate the analgesic fraction from the venom of Naja naja atra, and then to determine its properties by biochemical methods, such as SDS-polyacryamide gel electrophoresis ( SDS-PAGE), HPLC and Mole-toff. RESULTS: HPLC showed its relative purity was 95% (HPLC)and Mw was 6741. 236 Da. We observed that peripheral administration of neurotoxin strongly reduced the mechanical allogynia and thermal hyperalgesia for 24 hours, associated with this neuropathy (L5 spinal nerve ligation). CONCLUSION: The fraction from venom of Naja naja atra has significant analgesic effect and it is worth further developing.

[Study on anti-tumor effect of dry and fresh Gekko swinhonis freeze-dried powders on mice sarcoma S180 and acute toxicity testing of two powders].

OBJECTIVE: To investigate the anti-tumor activity of dry Gekko swinhonis freeze-dried powder (DGFP) and fresh G. swinhonis freeze-dried powder (FGFP) on mice sarcoma S180 and acute toxicity testing of the two powders. METHOD: Mice xenotransplant model of sarcoma S180 was established. Eighty mice were randomly divided into 8 groups. Control group were orally administrated by saline, another intraperitoneally injected with 5-Fu, the other six groups were orally administrated by DGFP and FGFP, each at three different doses (low, moderate and high). Rate of restraining tumor, index of thymus and spleen were calculated after 10 days' treatment. Acute toxicity testing tried to figure out LDs and LD, of DGFP and FGFP. RESULT: The restraining tumor rates of DGFP and FGFP each at three doses were 31.4%, 50.8%, 37.7% and 14.8%, 19.1%, 54.7%. DGFP and FGFP elevated the thymic weight and thymic index of the mice to different extent. There were no significant differences among the eight groups in their spleen weight and spleen index. Acute toxicity testing did not figure out LD50 of DGFP and FGFP. In LD0 test, the administrating dosages of DGFP and FGFP given to the mice were both more than 2000 times than those given to patients on clinic. The result showed nothing abnormal in DGFP group. Compared with the DGFP and control group there was only a significant body weight decrease (P < 0.01) in the FGFP group in the first three days. However, on the fifth day and the seventh day there was no significant difference. CONCLUSION: DGFP and FGFP have conspicuous anti-tumor effects in vivo. The mechanism may be related to the elevated cellular immune function. Acute toxicity testing reveals that DGFP and FGFP are quite safe for conventional oral use on clinic.

Circumventricular organ origin of domoic acid-induced neuropathology and toxicology.

The neuroexcitotoxin, domoic acid, was responsible for an episode of mussel poisoning in Eastern Canada in 1987. Severe neurologic impairment and some deaths occurred. We have characterized the nature of domoate-induced neuropathology in the mouse brain. Domoic acid was administered intraperitoneally at doses of 2, 3 or 7 mg/kg to Swiss-Webster mice. Brains were examined at 0.5, 1, 24, 48 or 72 h postinjection for evidence of damage. Significant pathologic changes occurred only after the largest dose of domoic acid. Damage was confined to circumventricular organs lacking a blood-brain barrier and their environs, including the organon vasculosum of the lamina terminalis, subfornical organ, mediobasal hypothalamus and area postrema. The neural damage induced by domoic acid was evident at as early as 30 min after injection and increased by 60 min postinjection. The loci of domoic acid-induced neuropathological changes accounts for several central and peripheral effects and toxicities observed following systemic domoate treatment, these included gastroduodenal lesions, hypodipsia, analgesia, and blood pressure fluctuations.

Comparative efficacy of two microdoses of a potentized homoeopathic drug, Cadmium Sulphoricum, in reducing genotoxic effects produced by cadmium chloride in mice: a time course study.

BACKGROUND: Cadmium poisoning in the environment has assumed an alarming problem in recent years. Effective antimutagenic agents which can reverse or combat cadmium induced genotoxicity in mice have not yet been reported. Therefore, in the present study, following the homeopathic principle of "like cures like", we tested the efficacy of two potencies of a homeopathic drug, Cadmium Sulphoricum (Cad Sulph), in reducing the genotoxic effects of Cadmium chloride in mice. Another objective was to determine the relative efficacy of three administrative modes, i.e. pre-, post- and combined pre and post-feeding of the homeopathic drugs. For this, healthy mice, Mus musculus, were intraperitoneally injected with 0.008% solution of CdCl2 @ 1 ml/100 gm of body wt (i.e. 0.8 mcg/gm of bw), and assessed for the genotoxic effects through such studies as chromosome aberrations (CA), micronucleated erythrocytes (MNE), mitotic index (MI) and sperm head anomaly (SHA), keeping suitable succussed alcohol fed (positive) and CdCl2 untreated normal (negative) controls. The CdCl2 treated mice were divided into 3 subgroups, which were orally administered with the drug prior to, after and both prior to and after injection of CdCl2 at specific fixation intervals and their genotoxic effects were analyzed. RESULTS: While the CA, MNE and SHA were reduced in the drug fed series as compared to their respective controls, the MI showed an apparent increase. The combined pre- and post-feeding of Cad Sulph showed maximum reduction of the genotoxic effects. CONCLUSIONS: Both Cad Sulph-30 and 200 were able to combat cadmium induced genotoxic effects in mice and that combined pre- and post-feeding mode of administration was found to be most effective in reducing the genotoxic effect of CdCl2 followed by the post-feeding mode.

Inhibition of (-)-propranolol hydrochloride by its enantiomer in white mice.

BACKGROUND: This study is based on the hypothesis, that the toxic or physiological effects of an optical isomer may be counteracted or reversed by the administration of a potentized preparation of one of its stereoisomers. In the present study the enantiomer was used. METHODS: 154 ICR conventional mice were used. 77 mice were administered (R)-(+)-propranolol HCl homeopathic potency prior to and during the experiment, and the other 77 were administered indistinguishable placebo. On the day of the experiment the mice were sedated with intraperitoneal Rometar. Once sedated they were injected intraperitoneally with the LD50 dose of (S)-(-)-propranolol HCl. RESULTS: The end point for statistical analysis was the difference in survival between the placebo and treatment mice. The odds ratio for survival of treatment mice relative to placebo mice was 1.64. The hypothesis of equal survival proportions gave a chi-square of 2.0916 (1 degree of freedom), which has a p-value of 0.1481. The analysis was then adjusted for mouse weight and intraperitoneal (-)-propranolol dosage using a logistic regression (LR) model. The LR treatment odds ratio was 2.017 and the LR treatment chi-square was 2.8864 (1 degree of freedom), which has a p-value of 0.0893. Consequently we accept the null hypothesis of no treatment effect on survival. The odds ratio estimates show that the treatment mice are 2.02 times more likely to survive than placebo mice, but this was not statistically significant with p = 0.089. Nine percent more treatment mice survived than placebo mice. The investigators accustomed to handling rodents noted that mouse recovery seemed substantially faster in the treatment mice than in the placebo mice.

Inhibition of (-)-trans-(1S,2S)-U50488 hydrochloride by its enantiomer in white mice -- a placebo-controlled, randomized study.

BACKGROUND: Previous studies have been performed to see if toxicity of optically active compounds may be inhibited by potentized preparations of their enantiomers. The present study is based on the hypothesis that the toxic effects of an optical isomer may be counteracted or reversed by the administration of a potentized preparation of one of its stereoisomers and in particular the enantiomer (patent applied for). METHODS: The design was prospective, blind, randomized, and placebo-controlled. 210 ICR conventional mice were used. 105 mice were administered a mixture of (+)-U50488 hydrochloride homeopathic potencies prior to and during the experiment, and the other 105 were administered indistinguishable placebo. The first 52 mice were used to establish an LD(50) of intraperitoneally administered (-)-U50488 hydrochloride under the conditions of this experiment. The estimated LD(50) was 25 mg/kg. The remaining 158 mice were then administered this LD(50) of (-)-U50488 HCl intraperitoneally. One mouse from the placebo group was excluded from the analysis because it died immediately after the possibly intravenous injection of (-)-U50488 HCl. RESULTS: 67% of homeopathy mice survived compared with 47% of placebo mice. The end point for statistical analysis was the difference in survival between the placebo and homeopathy mice. The analysis was adjusted for mouse weight using a logistic regression (LR) model. The LR treatment odds ratio for survival of treatment mice relative to placebo mice was 2.301 and the LR treatment chi-square was 6.2030 (1 degree of freedom), which has a p-value of 0.0128. Consequently, we reject the null hypothesis of no treatment effect on survival. CONCLUSION: We conclude that toxicity of intraperitoneal injection of (-)-U50488 hydrochloride may be inhibited by administration of a mixture of potencies of its enantiomer.

Inhibition of (-)-propranolol hydrochloride by its enantiomer in white mice--a placebo-controlled randomized study.

BACKGROUND: A previous pilot study was performed to see if toxicity of (S)-(-)-propranolol hydrochloride may be inhibited by a potentized preparation of its enantiomer. The present study is based on the hypothesis that the toxic effects of an optical isomer, may be counteracted or reversed by the administration of a potentized preparation of one of its stereoisomers, and in particular the enantiomer. METHODS: 508 ICR conventional mice were used. 254 mice were administered (R)-(+)-propranolol HCl homeopathic potency prior to and during the experiment, and the other 254 were administered indistinguishable placebo. On the day of the experiment mice were anesthetized with intraperitoneal Rometar. Once sedated the mice were administered the LD50 dose of (-)-propranolol HCl intraperitoneally. RESULTS: The end point for statistical analysis was the difference in survival between the placebo and treatment mice. The odds ratio for survival of treatment mice relative to placebo mice was 1.52. The hypothesis of equal survival proportions gave a chi-square of 5.0429 (1 degree of freedom), which has a p-value of 0.0247. The analysis was then adjusted for mouse weight and intraperitoneal (-)-propranolol dosage using a logistic regression (LR) model. The LR treatment odds ratio was 1.51 and the LR treatment chi-square was 4.8112 (1 degree of freedom), which has a p-value of 0.0283. Consequently, we reject the null hypothesis of no treatment effect on survival. Eleven percent more treatment mice survived than placebo mice. CONCLUSION: We conclude that the toxicity of intraperitoneal (-)-propranolol HCl, may be counteracted by administration of a potency of its enantiomer, in ICR conventional mice which have survived preceding intraperitoneal Rometar injection, and pre-dosing with (+)-propranolol HCl homeopathic potency.

The effect of the glucocorticoid Oradexon on endotoxin-induced peritoneal cell response.

Glucocorticoids are important modulators of immune reactions. They are capable of antagonising several effects of the bacterial endotoxin by inhibiting endotoxin-induced leukocyte activation, and the production of cytokines and inflammatory mediators. We earlier demonstrated that the antiglucocorticoid RU 38486 enhances the cytokine production induced by endotoxin and aggravates the course of experimental endotoxic and septic shock. In the present study we investigated the effect of the glucocorticoid Oradexon on the endotoxin-induced peritoneal cell response. For measurement of the peritoneal cell response, male CFLP mice (20-25 g) were injected i.p. with 10 microg/10 g body weight endotoxin (E. coli 026:B6 LPS, Difco Lab, Detroit, lot 110273JB). Dexamethasone (Oradexon, N.V, Organon Oss, The Netherlands) was administered i.p., i.v. or s.c. in a dose of 0.1 mg/10 g body weight, alone or concomitantly with endotoxin. We found that bacterial endotoxin increased the total cell count due to neutrophilia at 24 hours and, due to increases in the number of macrophages and lymphocytes 48 and 72 hours after treatment, respectively. The i.p., i.v., and s.c. injection of Oradexon, increased the total cell count and the macrophage count at 24, 48 and 72 hours. The i.p., s.c. and i.v. injection of Oradexon, concomitantly with endotoxin, reduced the total cell count at 48 and 72 hours, due to decreases in the macrophage count. The i.p., i.v. or s.c. administration of Oradexon concomitantly with LPS decreased the lymphocyte count and the neutrophil count at 24 and 72 hours. These results prove that glucocorticoids are capable of modifying the immune cell reactions induced by endotoxin.

Efecto del parathion sobre el índice de apoptosis en hepatocitos de ratones CF1 / Effect of parathion on the apoptosis of CF1 mouse hepatocytes

El las últimas décadas el uso masivo de agropesticidas órganofosforados, como Parathion y Malathion, ha permitido el control de plagas en la producción hortofrutícola, mejorando la productividad e incrementando la oferta de alimentos de mayor calidad. Sin embargo, pese a su efectividad, estos compuestos químicos son potenciales causantes de daños morfológicos y genéticos, de alto riesgo para la Salud Humana y animal (Draper, 1985; Rodríguez y Bustos-Obregón, 2000). El parathion© (PT), inhibidor de la aceltilcolinesterasa, se metaboliza en hígado, pulmón y cerebro. El efecto tóxico se debe a un proceso de desulfuración oxidativa hepática, que transforma el PT en paraoxon (PO), siendo éste su metabolito activo (Chambers y Chambers, 1990; Siller et al., 1997). El objetivo del presente trabajo es evaluar los efectos de una dosis única de PT sobre los índices de apoptosis en hepatocitos de ratón CFI. Se usaron ratones macho CFI de 8 a 10 semanas, con un peso promedio de 30 g, a los cuales se les aplicó una dosis intraperitoneal única de PT de 20 mg/Kg de peso (Sobarzo y Bustos-Obregón, 2000). Posteriormente fueron sacrificados a 1, 8, 16, 28 y 50 días postratamiento. El análisis histológico del hígado se realizó mediante microscopía óptica sobre cortes teñidos con hematoxilina/eosina en que se analizó la presencia y frecuencia de hepatocitos apoptóticos. Los resultados obtenidos permiten demostrar el efecto del PT sobre el hepatocito con un aumento estadísticamente significativo de apoptosis. Se postula que el PT es carcinogénico, que bloquea o modifica la capacidad de replicación de los hepatocitos, alterando la susceptibilidad del tejido hepático (Fausto, 2000; Metcalfe y Streuli, 1997). Se concluye que el PT tiene un efecto tóxico, aún en dosis consideradas bajas, aumentando significativamente los índices de eventos apoptóticos, alterando el ciclo celular y afectando la histofisiología del tejido hepático