Effect of Boschniakia rossica on expression of GST-P, p53 and p21(ras)proteins in early stage of chemical hepatocarcinogenesis and its anti-inflammatory activities in rats

AIM:To investigate the effect of Boschniakia rossica (BR) extract on expression of GST-P, p53 and p21(ras) proteins in early stage of chemical hepatocarcinogenesis in rats and its anti-inflammatory activities.METHODS:The expression of tumor marker-placental form glutathione S-transferase (GST-P), p53 and p21(ras) proteins were investigated by immunohisto-chemical techniques and ABC method. Anti-inflammatory activities of BR were studied by xylene and croton oil-induced mouse ear edema, carrageenin, histamine and hot scald-induced rat pow edema, adjuvant-induced rat arthritis and cotton pellet induced mouse granuloma formation methods.RESULTS:The 500mg/kg of BR-H2O extract frac-tionated from BR-Methanol extract had inhibitory effect on the formation of DEN-induced GST-P-positive foci in rat liver (GST-P staining was 78% positive in DEN+AAF group vs 20% positive in DEN+AAF+BR group, P<0.05) and the expression of mutant p53 and p21(ras) protein was lower than that of hepatic preneoplastic lesions (33% and 22% positive respectively in DEN+AAF group vs negative in DEN+AAF+BR group). Both CH(2)Cl(2) and H(2)O extracts from BR had anti-inflamatory effect in xylene and crotonoil induced mouse ear edema (inhibitory rates were 26%-29% and 35%-59%, respectively). BR H(2)O extract exhibited inhibitory effect in carrageenin, histamine and hot scald-induced hind paw edema and adjuvant-induced arthritis in rats and cotton pellet-induced granuloma formation in mice.CONCLUSION:BR extract exhibited inhibitory effect on formation of preneoplastic hepatic foci in early stage of rat chemical hepato-carcinogenesis.Both CH(2)Cl(2) and H(2)O extracts from BR exerted anti-inflammatory effect in rats and mice.     

Natural products and food components with anti-Helicobacter pylori activities

The bacterial pathogen Helicobacter pylori (H. pylori) colonizes in over half of the world’s population. H. pylori that establishes life-long infection in the stomach is definitely associated with gastro-duodenal diseases and a wide variety of non-gastrointestinal tract conditions such as immune thrombocytopenia. Triple therapy which consists of a proton pump inhibitor and combinations of two antibiotics (amoxicillin, clarithromycin or amoxicillin, metronidazol) is commonly used for H. pylori eradication. Recently, the occurrence of drug-resistant H. pylori and the adverse effect of antibiotics have severely weakened eradication therapy. Generally antibiotics induce the disturbance of human gastrointestinal microflora. Furthermore, there are inappropriate cases of triple therapy such as allergy to antibiotics, severe complications (liver and/or kidney dysfunction), the aged and people who reject the triple therapy. These prompt us to seek alterative agents instead of antibiotics and to develop more effective and safe therapy with these agents. The combination of these agents actually may result in lower a dose of antibiotics. There are many reports world-wide that non-antibiotic substances from natural products potentially have an anti-H. pylori agent. We briefly review the constituents derived from nature that fight against H. pylori in the literature with our studies.     

Anti-Helicobacter pylori activities of Chenopodium ambrosioides L. in vitro and in vivo

AIM: To investigate the bactericidal effects of Chenopodium ambrosioides L.(CAL) against Helicobacter pylori(H.pylori) both in vitro and in vivo.METHODS: For in vitro experiments, the inhibitory activity of CAL was tested using an agar dilution method; H.pylori strain NCTC11637 was incubated on Columbia blood agar plates containing serial concentrations of CAL.The minimal inhibitory concentration(MIC) was determined by the absence of H.pylori colonies on the agar plate.Time-kill curves were used to evaluate bactericidal activity; the average number of colonies was calculated at 0, 2, 8 and 24 h after liquid incubation with concentrations of CAL at 0.5, 1, and 2 × MIC.For in vivo experiments, H.pylori-infected mice were randomly divided into CAL, triple therapy(lansoprazole, metronidazole, and clarithromycin), blank control, or H.pylori control groups.The eradication ratios were determined by positive findings from rapid urease tests(RUTs) and by histopathology.RESULTS: In vitro, the MIC of CAL against H.pylori was 16 mg/L.The time-kill curves showed a stable and persistent decreasing tendency with increasing CAL concentration, and the intensity of the bactericidal effect was proportional to dose; the 1 and 2 × MIC completely inhibited the growth of H.pylori at 24 h.In vivo, the eradication ratios in the CAL group were60%(6/10) by RUT and 50%(5/10) by histopathology.Ratios in the triple therapy group were both 70%(7/10), and there was no difference between the CAL and triple therapy groups.Histopathologic evaluation revealed massive bacterial colonization on the surface of gastric mucosa and slight infiltration of mononuclear cells after inoculation with H.pylori, but no obvious inflammation or other pathologic changes in gastric mucosa of mice from CAL and triple therapy groups.CONCLUSION: CAL demonstrates effective bactericidal activity against H.pylori both in vitro and in vivo.     

Antinociceptive and anti-inflammatory activities of crude extracts of Ipomoea involucrata leaves in mice and rats

ObjectiveTo investigate antinociceptive and anti-inflammatory activities of crude extract from Ipomoea involucrata leaves (Convolvulaceae) in mice and rats.MethodsThe antinociceptive activity was tested using acetic acid-induced abdominal writhing test in mice. The anti-inflammatory activity was evaluated using egg albumin induced oedema of rat paw.ResultsPhytochemical screening showed the presence of alkaloids, flavonoids, saponins, terpenoids and tannin. At the doses of 25–100 mg/kg, Ipomoea involucrata exhibited dose-dependent and significant increase in pain threshold in acetic acid –induced writhing test of mice (P<0.05, student t-test) The administration of Ipomoea involucrata leaf extract (25–100 mg/kg) showed dose-dependent decreases in paw volume of egg albumin induced oedema in rats and a significant higher anti-inflammatory activity compared to the standard control (Aspirin).ConclusionsThese results support the claims on the traditional use of the of Ipomoea involucrata leaves in the treatment of toothache, rheumatic pains and other inflammatory conditions. Studies on the isolation and structural elucidation of the active principle are still needed being carried out.     

Probiotics improve survival of septic rats by suppressing conditioned pathogens in ascites

AIM: To investigate the benefits of probiotics treatment in septic rats. METHODS: The septic rats were induced by cecal ligation and puncture. The animals of control, septic model and probiotics treated groups were treated with vehicle and mixed probiotics, respectively. The mixture of probiotics included Bifidobacterium longum , Lacto-bacillus bulgaricus and Streptococcus thermophilus . We observed the survival of septic rats using different amounts of mixed probiotics. We also detected the bacterial population in ascites and blood of experimental sepsis using cultivation and real-time polymerase chain reaction. The severity of mucosal inflammation in colonic tissues was determined. RESULTS: Probiotics treatment improved survival of the rats significantly and this effect was dose dependent. The survival rate was 30% for vehicle-treated septic model group. However, 1 and 1/4 doses of probiotics treatment increased survival rate significantly comparedwith septic model group (80% and 55% vs 30%, P < 0.05). The total viable counts of bacteria in ascites decreased significantly in probiotics treated group compared with septic model group (5.20 ± 0.57 vs 9.81 ± 0.67, P < 0.05). The total positive rate of hemoculture decreased significantly in probiotics treated group compared with septic model group (33.3% vs 100.0%, P < 0.05). The population of Escherichia coli and Staphy-lococcus aureus in ascites of probiotics treated group were decreased significantly compared with that of septic model group (3.93 ± 0.73vs 8.80 ± 0.83,P < 0.05; 2.80 ± 1.04 vs 5.39 ± 1.21, P < 0.05). With probiotics treatment, there was a decrease in the scores of inflammatory cell infiltration into the intestinal mucosa in septic animals (1.50 ± 0.25vs 2.88 ± 0.14,P < 0.01). CONCLUSION: Escherichia coli and Staphylococcus aureus may be primary pathogens in septic rats. Probiotics improve survival of septic rats by suppressing these conditioned pathogens.     

Helicobacter pylori-negative,non-steroidal anti-inflammatory drug:Negative idiopathic ulcers in Asia

Since the discovery of Helicobacter pylori(H.pylori)infection in the stomach,the bacteria infection and non-steroidal anti-inflammatory drugs(NSAIDs)use had been considered to be the 2 main causes of peptic ulcers.However,there have been recent reports of an increase in the proportion of peptic ulcers without these known risk factors;these are termed idiopathic peptic ulcers.Such trend was firstly indicated in 1990s from some reports in North America.In Asia,numerous studies reported that idiopathic ulcers accounted for a small percentage of all ulcers in the 1990s,but in the2000s,multiple studies reported that the proportion of idiopathic ulcers had reached 10%-30%,indicating that the incidence of idiopathic ulcers in Asia has also been rising in recent years.While a decline in H.pylori infection rates of general population in Asia is seen as the main reason for the increased incidence of idiopathic ulcers,it is also possible that the absolute number of idiopathic ulcer cases has increased.Advanced age,serious systemic complication,and psychological stress are considered to be the potential risk factors for idiopathic ulcers.Management of idiopathic ulcers is challenging,at present,because there is no effective preventative measure against recurrence in contrast with cases of H.pylori-positive ulcers and NSAIDs-induced ulcers.As it is expected that H.pylori infection rates in Asia will decline further in the future,measures to treat idiopathic ulcers will also likely become more important.     

Testicular and epididymal pathology in Yankasa rams experimentally infected with Trypanosoma congolense

Objective

To investigate the pathological effect of experimental Trypanosoma congolense (T. congolense) infection on the testes and epididymis of Yankasa rams.

Methods

Nine intact un-castrated rams were obtained and divided into 2 groups of 6 infected with 1×10⁶ T. congolense and 3 uninfected controls. Four infected and one uninfected control rams were sacrificed on Day 75 post infection and the remaining four rams (two each of infected and control groups) on Day 86 post infection.

Results

All infected rams became parasitaemic within 7-11 days post infection with clinical trypanosomosis characterized by peri-orbital oedema and a transient period of scrotal oedema, this was followed by progressive decrease in scrotal size. At postmortem, the organs from infected rams were pale and emaciated but no observable lesion was seen in the uninfected control group. At histology, the testes showed areas of necrosis on the interstitial tissue characterized by destruction of cellular structures within the tissues and seminiferous tubules. There was moderate to severe testicular degeneration manifested by loss of tissue architecture and infiltration with macrophages, neutrophils, lymphocytes and plasma cells. Fibrous connective tissues were also seen in the testes of the infected rams indicative of replacement of normal cells by fibrous connective tissue. The epididymis of the infected rams had collection of some spermatocytes and desquamated epithelial cells in the epithelial ducts although others were devoid of spermatocytes in their ducts leading to loss of epididymal sperm reserves. However, these were not seen in the uninfected control rams.

Conclusions

T. congolense causes testicular and epididymal damage which may render the rams infertile or sterile.     

Interaction between Helicobacter pylori infection,nonsteroidal anti-inflammatory drugs and/or low-dose aspirin use: Old question new insights

Previous reports clearly demonstrated that Helicobacter pylori (H. pylori) infection, nonsteroidal anti-inflammatory drugs (NSAID) or low dose aspirin (ASA) use significantly and independently increased the risk for the development of peptic ulcer disease. Today, the presence of H. pylori infection associated with low dose ASA and/or NSAID use in the same patient is becoming more frequent and therefore the potential interaction between these factors and the consequences of it has important implications. Whether NSAID intake in the presence of H. pylori infection may further increase the risk of peptic ulcer carried by the presence of only one risk factor is still a matter of debate. Studies on the interaction between the two risk factors yielded conflicting data and no consensus has been reached in the last years. In addition, the interaction between H. pylori infection and low-dose ASA remains even more controversial. In real clinical practice, we can find different clinical scenarios involving these three factors associated with the presence of different gastrointestinal and cardiovascular risk factors. These huge variety of possible combinations greatly hinder the decision making process of physicians.     

Novel methylxanthine derivative-mediated anti-inflammatory effects in inflammatory bowel disease

Family 18 chitinases have a binding capacity with chitin, a polymer of N-acetylglucosamine. Recent studies strongly suggested that chitinase 3-like 1(CHI3L1, also known as YKL-40) and acidic mammalian chitinase, the two major members of family 18 chitinases, play a pivotal role in the pathogenesis of inflammatory bowel disease(IBD), bronchial asthma and several other inflammatory disorders. Based on the data from highthroughput screening, it has been found that three methylxanthine derivatives, caffeine, theophylline, and pentoxifylline, have competitive inhibitory effects against a fungal family 18 chitinase by specifically interacting with conserved tryptophans in the active site of this protein. Methylxanthine derivatives are also known as adenosine receptor antagonists, phosphodiesterase inhibitors and histone deacetylase inducers. Anti-in-flammatory effects of methylxanthine derivatives have been well-documented in the literature. For example, a beneficial link between coffee or caffeine consumption and type 2 diabetes as well as liver cirrhosis has been reported. Furthermore, theophylline has a long history of being used as a bronchodilator in asthma therapy, and pentoxifylline has an immuno-modulating effect for peripheral vascular disease. However, it is still largely unknown whether these methylxanthine derivativemediated anti-inflammatory effects are associated with the inhibition of CHI3L1-induced cytoplasmic signaling cascades in epithelial cells. In this review article we will examine the above possibility and summarize the biological significance of methylxanthine derivatives in intestinal epithelial cells. We hope that this study will provide a rationale for the development of methylxanthine derivatives, in particular caffeine,-based antiinflammatory therapeutics in the field of IBD and IBDassociated carcinogenesis.     

Effects of four Bifidobacteria on obesity in high-fat diet induced rats

AIM:To compare the effects of four Bifidobacteria strains(Bifidobacteria L66-5,L75-4,M13-4 and FS31-12,originated from normal human intestines) on weight gain,lipid metabolism,glucose metabolism in an obese murine model induced by high-fat diet.METHODS:Forty-eight Sprague-Dawley rats were randomly divided into six groups.Control group received standard chow,model group received high-fat diet,and intervention groups received high-fat diet added with different Bifidobacteria strains isolated from healthy volu...     

Tricyclic antidepressant imipramine reduces the insulin secretory rate in islet cells of Wistar albino rats through a calcium antagonistic action

Aims/hypothesis Treatments with antidepressants have been associated with modifications in glucose homeostasis. The aim of this study was to assess the effect of imipramine, a tricyclic antidepressant, on insulin-secreting cells.Methods Insulin radioimmunoassay, radioisotopic, fluorimetric and patch-clamp methods were used to characterise the effects of imipramine on ionic and secretory events in pancreatic islet cells from Wistar albino rats.Results Imipramine induced a dose-dependent decrease in glucose-stimulated insulin output (IC₅₀=5.2 µmol/l). It also provoked a concentration-dependent reduction in ⁴⁵Ca outflow from islets perifused in the presence of 16.7 mmol/l glucose. Moreover, imipramine inhibited the increase in ⁴⁵Ca outflow mediated by K⁺ depolarisation. Patch-clamp recordings further revealed that imipramine provoked a marked and reversible decrease of the inward Ca²⁺ current. In single islet cells, imipramine counteracted the rise in [Ca²⁺]_i evoked by glucose or high K⁺ concentrations.Conclusions/interpretation These data indicate that imipramine dose-dependently reduces the insulin secretory rate from rat pancreatic beta cells. Such an effect appears to be mediated by the inhibition of voltage-sensitive Ca²⁺ channels with subsequent reduction in Ca²⁺ entry. Thus, it is possible that some adverse effects of imipramine are related, at least in part, to its capacity to behave as a Ca²⁺ entry blocker.Abbreviations FOR fractional outflow rate     

EGLN1 involvement in high-altitude adaptation revealed through genetic analysis of extreme constitution types defined in Ayurveda

It is being realized that identification of subgroups within normal controls corresponding to contrasting disease susceptibility is likely to lead to more effective predictive marker discovery. We have previously used the Ayurvedic concept of Prakriti, which relates to phenotypic differences in normal individuals, including response to external environment as well as susceptibility to diseases, to explore molecular differences between three contrasting Prakriti types: Vata, Pitta, and Kapha. EGLN1 was one among 251 differentially expressed genes between the Prakriti types. In the present study, we report a link between high-altitude adaptation and common variations rs479200 (C/T) and rs480902 (T/C) in the EGLN1 gene. Furthermore, the TT genotype of rs479200, which was more frequent in Kapha types and correlated with higher expression of EGLN1, was associated with patients suffering from high-altitude pulmonary edema, whereas it was present at a significantly lower frequency in Pitta and nearly absent in natives of high altitude. Analysis of Human Genome Diversity Panel-Centre d'Etude du Polymorphisme Humain (HGDP-CEPH) and Indian Genome Variation Consortium panels showed that disparate genetic lineages at high altitudes share the same ancestral allele (T) of rs480902 that is overrepresented in Pitta and positively correlated with altitude globally (P < 0.001), including in India. Thus, EGLN1 polymorphisms are associated with high-altitude adaptation, and a genotype rare in highlanders but overrepresented in a subgroup of normal lowlanders discernable by Ayurveda may confer increased risk for high-altitude pulmonary edema.     

Effect of Astragalus complanatus flavonoid on anti-liver fibrosis in rats

AIM: To observe the anti-liver fibrosis effect of Astragalus complanatus flavonoids(ACF) in rats. METHODS: The liver fibrosis model in rats was established by injecting interperitoneally 0.2 mL/100 g 0.5% dimethylnitrosamine, thrice a week. Meanwhile, the rats were administered ACF(30, 60, 120 mg/kg) or colchicine(0.1 mg/kg) once a day for 1 mo. Serum N-propeptide of type Ⅰ procollagen(PINP) and type Ⅲ procollagen(PIIINP) was measured using ELISA. Malondialdehyde(MDA) and superoxide dismutase(SOD) in hepatic tissue were evaluated. Matrix metal protease-1(MMP-1) mRNA expression was assayed by RT-PCR and the protein expression of tissue inhibitor of metal protease-1(TTMP-1) was analyzed by immunohistochemistry. RESULTS: In the ACF groups, SOD activity increased and MDA content decreased in comparison to the liver fibrosis model group. The serum PINP and PIIINP contents in ACF-2 and -3 group decreased compared to those in model group. In ACF-2 and -3 group, the expression of MMP-1 mRNA increased significantly and the protein expression of TTMP-1 decreased compared to that in model group. CONCLUSION: The antifibrotic mechanisms of ACF are associated with its influence on lipid peroxidation and collagen synthesis and degradation.     

Effects of the viability of Lactobacillus rhamnosus GG on rotavirus infection in neonatal rats

AIM: To study the effects of live and dead Lactobacillus rhamnosus GG (GG) on rotavirus infection in a neonatal rat model.METHODS: At the age of 2 d, suckling Lewis rat pups were supplemented with either live or dead GG and the treatment was continued daily throughout the experiment. At the age of 5 and 6 d the pups received oral rotavirus (RV) SA-11 strain. The pups were sacrificed at the age of 7 or 8 d by decapitation. The gastrointestinal tract was removed and macroscopic observations were done. The consistency of feces in the colon was classified using a four-tier system. RV was detected from the plasma, small intestine, colon and feces by real-time quantitative polymerase chain reaction (PCR).RESULTS: In this neonatal rat model, RV induced a mild-to-moderate diarrhea in all except one pup of the RV-inoculated rats. RV moderately reduced body weight development from day 6 onwards. On day 7, after 2 d of RV infection, live and dead GG groups gained significantly more weight than the RV group without probiotics [36% (P = 0.001) and 28% (P = 0.031), respectively]. In addition, when compared with the RV control group, both live and dead GG reduced the weight ratio of colon/animal body weight to the same level as in the healthy control group, with reductions of 22% (P = 0.002) and 28% (P < 0.001), respectively. Diarrhea increased moderately in both GG groups. However, the diarrhea incidence and severity in the GG groups were not statistically significantly different as compared with the RV control group. Moreover, observed diarrhea did not provoke weight loss or death. The RV control group had the largest amount of RV PCR-positive samples among the RV-infected groups, and the live GG group had the smallest amount. Rats receiving live GG had significantly less RV in the colon (P = 0.027) when compared with the RV control group. Live GG was also more effective over dead GG in reducing the quantity of RV from plasma (P = 0.047).CONCLUSION: Both live and dead GG have beneficial effects in RV infection. GG may increase RV clearance from the body and reduce colon swelling.     

Influence of streptozotocin and alloxan induced diabetes on the metabolism of dermal collagen in albino rats

Summary The metabolism of collagen in male rats made diabetic by treatment with either streptozotocin or alloxan was studied after the injection of³H-proline by estimating specific and total³H-hydroxyproline activity in skin collagen fractions and urine. Experimentally induced diabetes was found to decrease the neutral salt-soluble and acid-soluble collagen with no change in insoluble collagen as compared to a control group. The specific and total radioactivity of³H-hydroxyproline in soluble and insoluble collagen fractions were also decreased. Studies of total³H-hydroxyproline activities in soluble collagens and insoluble collagen showed that the conversion of soluble to insoluble collagen was influenced by diabetes. Both streptozotocin and alloxan were found to increase urinary excretion of total hydroxyproline and³H-hydroxyproline during the first 12 h after the administration of³H-proline. Weekly analyses of urinary hydroxyproline also indicated a similar pattern. The results of the present investigation clearly indicate decreased synthesis and increased catabolism of collagen accompanied by accelerated conversion of soluble to insoluble collagen in experimentally induced diabetes.     

Analgesic and anti—inflammatory activities of the ethanol extract of the leaves of Helianthus Annus in Wistar rats

ObjectiveTo investigate the anti-inflammatory and analgesic effects of the ethanol extract of leaves of Helianthus annus L. (H. annus) in acclimatized Wistar rats.MethodsIt was undertaken using the albumin induced paw edema model of inflammation as well as both the hotplate and tail immersion analgesic test methods. Doses of the extract tested in experimental rats were 0.5 g/kg, 2 g/kg and 4 g/kg while negative and positive control rats received distilled water and indomethacin respectively.ResultsIt was shown that treatment with the tested doses of the extract effectively inhibited paw edema induced by egg albumin. This effect was comparable if not better than the observations made in rats treated with 10 mg/kg of indomethacin orally. Treatment with the extract was also observed to have significantly increased the mean tolerance time of rats to thermal noxious stimuli compared to control animals that had distilled water and appeared to be more effective than 10 mg/kg of indomethacin treatment.ConclusionsThese observations confirmed the presence of a strong anti-inflammatory and anti-noiciceptive activity in the ethanol extract of the leaves of H. annus and therefore validated the folkloric use of the leaves of this plant in treatment of pro-inflammatory, post traumatic situations.