AIM： To investigate the intestinal anti-inflammatory effect and mechanism of a probiotic Lactobacillus salivarius ssp. salivariusCECT5713 in the TNBS model of rat colitis. METHODS： Female Wistar rats （180-200 g） w...AIM： To investigate the intestinal anti-inflammatory effect and mechanism of a probiotic Lactobacillus salivarius ssp. salivariusCECT5713 in the TNBS model of rat colitis. METHODS： Female Wistar rats （180-200 g） were used in this study. A group of rats were administered orally the probiotic L. salivarius ssp. salivarius （5 × 10^8 CFU suspended in 0.5 mL of skimmed milk） daily for 3 wk. Two additional groups were used for reference, a non-colitic and a control colitic without probiotic treatment, which received orally the vehicle used to administer the probiotic. Two weeks after starting the experiment, the rats were rendered colitic by intracolonic administration of 10 mg of TNBS dissolved in 0.25 mL of 500 mL/L ethanol. One week after colitis induction, all animals were killed and colonic damage was evaluated both histologically and biochemically. The biochemical studies performed in colonic homogenates include determination of myeloperoxidase （MPO） activity, glutathione （GSH） content, leukotriene B4 （LTB4） and tumor necrosis factor α （TNF-α） levels, as well as inducible nitric oxide synthase （iNOS） expression. In addition, the luminal contents obtained from colonic samples were used for microbiological studies, in order to determine Lactobacilli and Bifidobacteria counts. RESULTS： Treatment of colitic rats with L. salivarius ssp. salivarius resulted in amelioration of the inflammatory response in colitic rats, when compared with the corresponding control group without probiotic treatment. This antiinflammatory effect was evidenced macroscopically by a significant reduction in the extent of colonic necrosis and/or inflammation induced by the administration of TNBS/ethanol （2.3±0.4 cm vs3.4±0.3 cm in control group, P〈0.01） and histologically by improvement of the colonic architecture associated with a reduction in the neutrophil infiltrate in comparison with non-treated colitic rats. The latter was confirmed biochemically by a significant reduction of colonic MPO activity （105.3±26.0 U/g vs 180.6±21.9 U/g, P〈0.05）, a marker of neutrophil infiltration. The beneficial effect was associated with an increase of the colonic GSH content （1 252±42 nmol/g vs i 087±51 nmol/g, P〈0.05）, which is depleted in colitic rats, as a consequence of the oxidative stress induced by the inflammatory process. In addition, the treatment of colitic rats with L. salivarius resulted in a significant reduction of colonic TNF-（~ levels （509.4±68.2 pg/g vs782.9±60.1 pg/g, P〈0.01） and in a lower colonic iNOS expression, when compared to TNBS control animals without probiotic administration. Finally, treated colitic rats showed higher counts of Lactobacilli species in colonic contents than control colitic rats, whereas no differences were observed in Bifidobacteria counts. CONCLUSION： Administration of the probiotic L. salivarius ssp. salivarius CECT5713 facilitates the recovery of the inflamed tissue in the TNBS model of rat colitis, an effect associated with amelioration of the production of some of the mediators involved in the inflammatory response in the intestine, such as cytokines, including TNF-α and NO. This beneficial effect could be ascribed to its effect on the altered immune response that occurs in this inflammatory condition.展开更多
In this study, Lactobacillus salivarius was isolated from human saliva by culture technique using MRS medium, identified through relevant biochemical protocols, and compared to a known control of Lactobacillus acidoph...In this study, Lactobacillus salivarius was isolated from human saliva by culture technique using MRS medium, identified through relevant biochemical protocols, and compared to a known control of Lactobacillus acidophilus sourced from Puritan Pride Inc., Qadelade, New York, USA. Primary isolation of microorganisms of interest was carried out at Cheznik Diagnostic and Research laboratories while animal preparation and treatment were carried out at Animal Farm, Mgbakwu Awka. Twenty wistar rats were used as the test animals and another twenty for positive control. The animals in the test group were induced to diarrhea using 1 ml of castor oil following 12-hour fasting while the control group was fed with daily feed and distilled water. The test animals upon production of diarrhoeic stool were fed with 1 × 10<sup>9</sup> cfu/ml (410 mg/dl w/v) of the Lactobacillus sp isolate in distilled water. The diarrhoea was resolved within 24 hours of treatment. The average weights of the animals taken just after preparation and early treatment (T<sub>x</sub>) showed appreciable loss in weight among the test animals possibly due to stress compared to initial weight at T<sub>0</sub>. However, upon continued treatment with the isolate, obvious weight gain (Tf) compared to T<sub>x</sub> was observed. The results of the haematological data at the final analysis showed proximal values and consistency to that of the control group following administration of the Lactobacillus sp isolate. The routine general health data of the test animals showed marked improvement upon treatment with the isolate. A test of significance at 0.05 showed a positive agreement (p Lactobacillus sp showed marked sensitivity to erythromycin, ceftazidine, and chloromphenicol antimicrobial drugs just like other known probiotic strains. No known health hazard or death was recorded. From the study, it was observed that saliva can also proffer a good site in addition to the gut and ileum for the isolation of probiotic microorganisms. It has also shown that the presumptive Lactobacillus salivarius, though strain-specific, possesses probiotic properties effective in resolving most gut issues but especially in diarrhoeic cases.展开更多
基金Supported by the Spanish Ministry of Science and Technology, No. SAF2002-02592 and by Institute de Salud 'Carlos Ⅲ', No. PI021732, with Funds from the European Union, and by Junta de Andalucia (CTS 164) Monica Comalada is a recipient of Juan de la Cierva Program from Spanish Ministry of Science and Technology. Laura Peran is a Recipient From Puleva Foundation Spain
文摘AIM： To investigate the intestinal anti-inflammatory effect and mechanism of a probiotic Lactobacillus salivarius ssp. salivariusCECT5713 in the TNBS model of rat colitis. METHODS： Female Wistar rats （180-200 g） were used in this study. A group of rats were administered orally the probiotic L. salivarius ssp. salivarius （5 × 10^8 CFU suspended in 0.5 mL of skimmed milk） daily for 3 wk. Two additional groups were used for reference, a non-colitic and a control colitic without probiotic treatment, which received orally the vehicle used to administer the probiotic. Two weeks after starting the experiment, the rats were rendered colitic by intracolonic administration of 10 mg of TNBS dissolved in 0.25 mL of 500 mL/L ethanol. One week after colitis induction, all animals were killed and colonic damage was evaluated both histologically and biochemically. The biochemical studies performed in colonic homogenates include determination of myeloperoxidase （MPO） activity, glutathione （GSH） content, leukotriene B4 （LTB4） and tumor necrosis factor α （TNF-α） levels, as well as inducible nitric oxide synthase （iNOS） expression. In addition, the luminal contents obtained from colonic samples were used for microbiological studies, in order to determine Lactobacilli and Bifidobacteria counts. RESULTS： Treatment of colitic rats with L. salivarius ssp. salivarius resulted in amelioration of the inflammatory response in colitic rats, when compared with the corresponding control group without probiotic treatment. This antiinflammatory effect was evidenced macroscopically by a significant reduction in the extent of colonic necrosis and/or inflammation induced by the administration of TNBS/ethanol （2.3±0.4 cm vs3.4±0.3 cm in control group, P〈0.01） and histologically by improvement of the colonic architecture associated with a reduction in the neutrophil infiltrate in comparison with non-treated colitic rats. The latter was confirmed biochemically by a significant reduction of colonic MPO activity （105.3±26.0 U/g vs 180.6±21.9 U/g, P〈0.05）, a marker of neutrophil infiltration. The beneficial effect was associated with an increase of the colonic GSH content （1 252±42 nmol/g vs i 087±51 nmol/g, P〈0.05）, which is depleted in colitic rats, as a consequence of the oxidative stress induced by the inflammatory process. In addition, the treatment of colitic rats with L. salivarius resulted in a significant reduction of colonic TNF-（~ levels （509.4±68.2 pg/g vs782.9±60.1 pg/g, P〈0.01） and in a lower colonic iNOS expression, when compared to TNBS control animals without probiotic administration. Finally, treated colitic rats showed higher counts of Lactobacilli species in colonic contents than control colitic rats, whereas no differences were observed in Bifidobacteria counts. CONCLUSION： Administration of the probiotic L. salivarius ssp. salivarius CECT5713 facilitates the recovery of the inflamed tissue in the TNBS model of rat colitis, an effect associated with amelioration of the production of some of the mediators involved in the inflammatory response in the intestine, such as cytokines, including TNF-α and NO. This beneficial effect could be ascribed to its effect on the altered immune response that occurs in this inflammatory condition.
基金This work was supported by funds from the Department of Science and Technology,Guangdong Province(2010B020313001)The Earmarked Fund for China Agriculture Research System(CARS-36),Ministry of Agriculture,PR China.
文摘In this study, Lactobacillus salivarius was isolated from human saliva by culture technique using MRS medium, identified through relevant biochemical protocols, and compared to a known control of Lactobacillus acidophilus sourced from Puritan Pride Inc., Qadelade, New York, USA. Primary isolation of microorganisms of interest was carried out at Cheznik Diagnostic and Research laboratories while animal preparation and treatment were carried out at Animal Farm, Mgbakwu Awka. Twenty wistar rats were used as the test animals and another twenty for positive control. The animals in the test group were induced to diarrhea using 1 ml of castor oil following 12-hour fasting while the control group was fed with daily feed and distilled water. The test animals upon production of diarrhoeic stool were fed with 1 × 10<sup>9</sup> cfu/ml (410 mg/dl w/v) of the Lactobacillus sp isolate in distilled water. The diarrhoea was resolved within 24 hours of treatment. The average weights of the animals taken just after preparation and early treatment (T<sub>x</sub>) showed appreciable loss in weight among the test animals possibly due to stress compared to initial weight at T<sub>0</sub>. However, upon continued treatment with the isolate, obvious weight gain (Tf) compared to T<sub>x</sub> was observed. The results of the haematological data at the final analysis showed proximal values and consistency to that of the control group following administration of the Lactobacillus sp isolate. The routine general health data of the test animals showed marked improvement upon treatment with the isolate. A test of significance at 0.05 showed a positive agreement (p Lactobacillus sp showed marked sensitivity to erythromycin, ceftazidine, and chloromphenicol antimicrobial drugs just like other known probiotic strains. No known health hazard or death was recorded. From the study, it was observed that saliva can also proffer a good site in addition to the gut and ileum for the isolation of probiotic microorganisms. It has also shown that the presumptive Lactobacillus salivarius, though strain-specific, possesses probiotic properties effective in resolving most gut issues but especially in diarrhoeic cases.
