AIM： To elucidate the mechanism of liver protection by inhibition of Kupffer cells （KCs） function.METHODS： All the animals were randomly divided into three groups. Blockade group （gadolinium chloride solution （G...AIM： To elucidate the mechanism of liver protection by inhibition of Kupffer cells （KCs） function.METHODS： All the animals were randomly divided into three groups. Blockade group （gadolinium chloride solution （GdCl3） injection plus ischemia/reperfusion （I/R） injury）：GdCl3 solution was injected once every 24 h for 2 d via the tail vein before I/R injury. Non-blockade group （saline solution injection plus I/R injury）： saline instead of GdCl3 as a control was injected as in the blockade group. Sham group： saline was injected without I/R injury. Liver samples were collected 4 h after blood inflow restoration. The blockade of the function of KCs was verified by immunostaining with an anti-CD68 mAb. Toll-like receptor2 （TLR2） was immunostained with a goat antimouse polydonal anti-TLR2 antibody. Membrane proteins were extracted from the liver samples and TLR2 protein was analyzed by Western blot. Portal vein serum and plasma were taken respectively at the same time point for further detection of the levels of tumor necrosis factor-α （TNF-α） and alanine aminotransferase （ALT）, an indicator of liver function.I/R injury via downregulation of theexpression of TLR2.RESULTS： Compared to non-blockade group, CD^68＋ cells significantly reduced in blockade group （OPTDI, optical density integral）： 32.97±10.55 vs 185.65±21.88, P〈0.01） and the liver function impairment was relieved partially （level of ALT： 435.89±178.37 U/L vs 890.21±272.91 U/L,P〈0.01）. The expression of TLR2 protein in blockade group significantly decreased compared to that in non- group（method of immunohistochemistry, OPDTI： 75.74±17.44 vs 170.58±25.14, P〈0.01; method of Western blot, A value： 125.89±15.49 vs 433.91±35.53, P〈0.02）. The latter correlated with the variation of CD68 staining （r = 0.745,P〈0.05）. Also the level of portal vein TNF-α decreased in blockade group compared to that in non-blockade group （84.45±14.73 ng/L vs 112.32±17.56 ng/L, P〈0.05）, butwas still higher than that in sham group （84.45±14.73 ng/L vs 6.07±5.33 ng/L, P〈0.01）.CONCLUSION： Inhibition of the function of KCs may protect liver against I/R injury via downregulation of the expression of TLR2.展开更多
AIM： To evaluate the effects of gliadin on the oxidative environment in the “in vivo-like”model of a three-dimensional cell culture system. METHODS： LoVo cell line （intestinal adenocarcinoma） multicellular spher...AIM： To evaluate the effects of gliadin on the oxidative environment in the “in vivo-like”model of a three-dimensional cell culture system. METHODS： LoVo cell line （intestinal adenocarcinoma） multicellular spheroids were treated with digested gliadin （with albumin used as a control）. Spheroid volumes, cell viability and morphology, lactate dehydrogenase （LDH） release, content of reduced glutathione （GSH） and activity of GSH-related enzymes were examined. The data were statistically analyzed using the Student＇s t-test （P〈0.05）. was considered statistically significant. RESULTS： Gliadin reduced cell viability （from 20% to 60%） and led to morphological alterations characterized by apoptotic findings and cytoskeletal injuries. LDH activity increased. The content of GSH reduced （-20% vscontrols）, and activity of GSH-related enzymes was significantly inhibited. CONCLUSION： Gliadin treatment induces an imbalance in the antioxidative mechanism of cells cultured by the three-dimensional technique. This alteration may explain cell damage directly caused by gliadin and the subsequentmorphological abnormalities.展开更多
OBJECTIVE： To evaluate the effects of Xuebijing （XBJ） injection in heat stroke （HS） rats and to inves- tigate the mechanisms underlying these effects. METHODS： Sixty anesthetized rats were random- ized into thre...OBJECTIVE： To evaluate the effects of Xuebijing （XBJ） injection in heat stroke （HS） rats and to inves- tigate the mechanisms underlying these effects. METHODS： Sixty anesthetized rats were random- ized into three groups and intravenously injected twice daily for 3 days with 4 mL XBJ （XBJ group） or phosphate buffered saline （HS and Sham groups） per kg body weight. HS was initiated in the HS and XBJ groups by placing rats in a simulated climate chamber （ambient temperature 40℃：, humidity 60% ）. Rectal temperature, aterial pressure, and heart rate were monitored and recorded. Time to HS onset and survival were determined, and serum concentrations of tumor necrosis factor （TNF）-α, in-terleukin （IL）-1β, IL-6, alanine-aminotransferase （ALT）, and aspartate-aminotransferase （AST） were measured. Hepatic tissue was harvested for patho- logical examination and electron microscopic ex- amination. Kupffer cells （KCs） were separated from liver at HS initiation, and the concentrations of se- creted TNF-α, IL-1β3 and IL-6 were measured. RESULTS： Time to HS onset and survival were signif- icantly longer in the XBJ than in the HS group. Moreover, the concentrations of TNF-α, IL-1β, IL-6, ALT and AST were lower and liver injury was milder in the XBJ than in the HS group. Heat-stress in- duced structural changes in KCs and hepatic cells were more severe in the HS than in the XBJ group and the concentrations of TNF-α, IL-1β and IL-6 se- creted by KCs were lower in the XBJ than in the HS group. CONCLUSION： XBJ can alleviate HS-induced sys- temic inflammatory response syndrome and liver injury in rats, and improve outcomes. These protec- tive effects may be due to the ability of XBJ to inhib- it cytokine secretion by KCs.展开更多
Objective： To promote stem cells differentiation into neurons and enhance neuromotor function after brain injury through brain-derived neurotrophic factor （BDNF） induction. Methods： Recombinant adenovirus vector ...Objective： To promote stem cells differentiation into neurons and enhance neuromotor function after brain injury through brain-derived neurotrophic factor （BDNF） induction. Methods： Recombinant adenovirus vector was applied to the transfection of BDNF into human-derived umbilical cord mesenchymal stem cells （UCMSCs）. Enzyme linked immunosorbent assay （ELISA） was used to determine the secretion phase of BDNF. The brain injury model of athymic mice induced by hydraulic pressure percussion was established for transplantation of stem cells into the edge of injury site. Nerve function scores were obtained, and the expression level of transfected and non-transfected BDNF, proportion of neuron specific enolase （NSE） and glial fibrillary acidic protein （GFAP）, and the number of apoptosis cells were compared respectively. Results： The BDNF expression achieved its stabilization at a high level 72 hours after gene transfection. The mouse obtained a better score of nerve function, and the proportion of the NSE-positive cells increased significantly （P〈0.05）, but GFAP-positive cells decreased in BDNF- UCMSCs group compared with the other two groups （P〈0.05）. At the site of high expression of BDNF, the number of apoptosis cells decreased markedly. Conclusion： BDNF gene can promote the differentiation of the stem cells into neurons rather than glial cells, and enhance neuromotor function after brain injury.展开更多
基金Supported by the National Natural Science Foundation of China, No. 30200272
文摘AIM： To elucidate the mechanism of liver protection by inhibition of Kupffer cells （KCs） function.METHODS： All the animals were randomly divided into three groups. Blockade group （gadolinium chloride solution （GdCl3） injection plus ischemia/reperfusion （I/R） injury）：GdCl3 solution was injected once every 24 h for 2 d via the tail vein before I/R injury. Non-blockade group （saline solution injection plus I/R injury）： saline instead of GdCl3 as a control was injected as in the blockade group. Sham group： saline was injected without I/R injury. Liver samples were collected 4 h after blood inflow restoration. The blockade of the function of KCs was verified by immunostaining with an anti-CD68 mAb. Toll-like receptor2 （TLR2） was immunostained with a goat antimouse polydonal anti-TLR2 antibody. Membrane proteins were extracted from the liver samples and TLR2 protein was analyzed by Western blot. Portal vein serum and plasma were taken respectively at the same time point for further detection of the levels of tumor necrosis factor-α （TNF-α） and alanine aminotransferase （ALT）, an indicator of liver function.I/R injury via downregulation of theexpression of TLR2.RESULTS： Compared to non-blockade group, CD^68＋ cells significantly reduced in blockade group （OPTDI, optical density integral）： 32.97±10.55 vs 185.65±21.88, P〈0.01） and the liver function impairment was relieved partially （level of ALT： 435.89±178.37 U/L vs 890.21±272.91 U/L,P〈0.01）. The expression of TLR2 protein in blockade group significantly decreased compared to that in non- group（method of immunohistochemistry, OPDTI： 75.74±17.44 vs 170.58±25.14, P〈0.01; method of Western blot, A value： 125.89±15.49 vs 433.91±35.53, P〈0.02）. The latter correlated with the variation of CD68 staining （r = 0.745,P〈0.05）. Also the level of portal vein TNF-α decreased in blockade group compared to that in non-blockade group （84.45±14.73 ng/L vs 112.32±17.56 ng/L, P〈0.05）, butwas still higher than that in sham group （84.45±14.73 ng/L vs 6.07±5.33 ng/L, P〈0.01）.CONCLUSION： Inhibition of the function of KCs may protect liver against I/R injury via downregulation of the expression of TLR2.
基金Supported by the San Paolo Foundation grant to "Centro per lo Studio della Celiachia"
文摘AIM： To evaluate the effects of gliadin on the oxidative environment in the “in vivo-like”model of a three-dimensional cell culture system. METHODS： LoVo cell line （intestinal adenocarcinoma） multicellular spheroids were treated with digested gliadin （with albumin used as a control）. Spheroid volumes, cell viability and morphology, lactate dehydrogenase （LDH） release, content of reduced glutathione （GSH） and activity of GSH-related enzymes were examined. The data were statistically analyzed using the Student＇s t-test （P〈0.05）. was considered statistically significant. RESULTS： Gliadin reduced cell viability （from 20% to 60%） and led to morphological alterations characterized by apoptotic findings and cytoskeletal injuries. LDH activity increased. The content of GSH reduced （-20% vscontrols）, and activity of GSH-related enzymes was significantly inhibited. CONCLUSION： Gliadin treatment induces an imbalance in the antioxidative mechanism of cells cultured by the three-dimensional technique. This alteration may explain cell damage directly caused by gliadin and the subsequentmorphological abnormalities.
基金Supported by the National Natural Science Foundation of China (Grant No. 81101406,81071529)the Project of Medical Research of PLA BWS12J108
文摘OBJECTIVE： To evaluate the effects of Xuebijing （XBJ） injection in heat stroke （HS） rats and to inves- tigate the mechanisms underlying these effects. METHODS： Sixty anesthetized rats were random- ized into three groups and intravenously injected twice daily for 3 days with 4 mL XBJ （XBJ group） or phosphate buffered saline （HS and Sham groups） per kg body weight. HS was initiated in the HS and XBJ groups by placing rats in a simulated climate chamber （ambient temperature 40℃：, humidity 60% ）. Rectal temperature, aterial pressure, and heart rate were monitored and recorded. Time to HS onset and survival were determined, and serum concentrations of tumor necrosis factor （TNF）-α, in-terleukin （IL）-1β, IL-6, alanine-aminotransferase （ALT）, and aspartate-aminotransferase （AST） were measured. Hepatic tissue was harvested for patho- logical examination and electron microscopic ex- amination. Kupffer cells （KCs） were separated from liver at HS initiation, and the concentrations of se- creted TNF-α, IL-1β3 and IL-6 were measured. RESULTS： Time to HS onset and survival were signif- icantly longer in the XBJ than in the HS group. Moreover, the concentrations of TNF-α, IL-1β, IL-6, ALT and AST were lower and liver injury was milder in the XBJ than in the HS group. Heat-stress in- duced structural changes in KCs and hepatic cells were more severe in the HS than in the XBJ group and the concentrations of TNF-α, IL-1β and IL-6 se- creted by KCs were lower in the XBJ than in the HS group. CONCLUSION： XBJ can alleviate HS-induced sys- temic inflammatory response syndrome and liver injury in rats, and improve outcomes. These protec- tive effects may be due to the ability of XBJ to inhib- it cytokine secretion by KCs.
基金This study was supported by the Project of Science and Technology Funds of Tianjin （No.06YFSZSF01200） and the National Natural Science Foundation of China （No. 30872668）
文摘Objective： To promote stem cells differentiation into neurons and enhance neuromotor function after brain injury through brain-derived neurotrophic factor （BDNF） induction. Methods： Recombinant adenovirus vector was applied to the transfection of BDNF into human-derived umbilical cord mesenchymal stem cells （UCMSCs）. Enzyme linked immunosorbent assay （ELISA） was used to determine the secretion phase of BDNF. The brain injury model of athymic mice induced by hydraulic pressure percussion was established for transplantation of stem cells into the edge of injury site. Nerve function scores were obtained, and the expression level of transfected and non-transfected BDNF, proportion of neuron specific enolase （NSE） and glial fibrillary acidic protein （GFAP）, and the number of apoptosis cells were compared respectively. Results： The BDNF expression achieved its stabilization at a high level 72 hours after gene transfection. The mouse obtained a better score of nerve function, and the proportion of the NSE-positive cells increased significantly （P〈0.05）, but GFAP-positive cells decreased in BDNF- UCMSCs group compared with the other two groups （P〈0.05）. At the site of high expression of BDNF, the number of apoptosis cells decreased markedly. Conclusion： BDNF gene can promote the differentiation of the stem cells into neurons rather than glial cells, and enhance neuromotor function after brain injury.
