AIM:To investigate the change in intestinal dendritic cell(DC)number in fulminant hepatic failure(FHF).METHODS:An animal model of FHF was created.Intestinal CD11b/c was detected by immunohistochemistry and Western blo...AIM:To investigate the change in intestinal dendritic cell(DC)number in fulminant hepatic failure(FHF).METHODS:An animal model of FHF was created.Intestinal CD11b/c was detected by immunohistochemistry and Western blot.Quantitative real-time polymerase chain reaction(PCR)was used to detect intestinal integrin-αm RNA expression.Intestinal CD83,CD86,CD74,CD3 and AKT were detected by immunohistochemistry,Western blot and PCR.Phosphorylated-AKT(p-AKT)was detected by immunohistochemistry and Western blot.RESULTS:In the FHF group[D-galactosamine(D-Galn)+lipopolysaccharide(LPS)group],the mice began to die after 6 h;conversely,in the D-Galn and LPS groups,the activity of mice was poor,but there were no deaths.Immunohistochemistry results showed that in FHF,the expression of CD11b/c(7988400±385941vs 1102400±132273,P<0.05),CD83(13875000±467493 vs 9257600±400364,P<0.05),CD86(7988400±385941 vs 1102400±13227,P<0.05)and CD74(11056000±431427 vs 4633400±267903,P<0.05)was significantly increased compared with the normal saline(NS)group.Compared with the NS group,the protein expression of CD11b/c(5.4817±0.77 vs 1.4073±0.37,P<0.05)and CD86(4.2673±0.69 vs 1.1379±0.42,P<0.05)was significantly increased.Itg-α(1.1224±0.3 vs 0.4907±0.19,P<0.05),CD83(3.6986±0.40 vs 1.0762±0.22,P<0.05)and CD86(1.5801±0.32 vs 0.8846±0.10,P<0.05)m RNA expression was increased significantly in the FHF group.At the protein level,expression of CD74in the FHF group(2.3513±0.52)was significantly increased compared with the NS group(1.1298±0.33),whereas in the LPS group(2.3891±0.47),the level of CD74 was the highest(P<0.05).At the gene level,the relative expression of CD74 m RNA in the FHF group(1.5383±0.26)was also significantly increased in comparison to the NS group(0.7648±0.22;P<0.05).CD3 expression was the highest in the FHF group(P<0.05).In the FHF,LPS and D-Galn groups,the expression of AKT at the protein and m RNA levels was elevated compared with the NS group,but there wasno statistical significance(P>0.05).The p-AKT protein expression in the FHF(1.54±0.06),LPS(1.56±0.05)and D-Galn(1.29±0.03)groups was higher than that in the NS group(1.07±0.03)(P<0.05).CONCLUSION:In FHF,a large number of DCs mature,express CD86,and activate MHC classⅡmolecular pathways to induce a T cell response,and the AKT pathway is activated.展开更多
Objective:Sertoli cells(SCs)provide physical support and material supply for germ cells and participate in the formation of blood-testis barrier.The number of SCs is directly proportional to the number of germ cells.A...Objective:Sertoli cells(SCs)provide physical support and material supply for germ cells and participate in the formation of blood-testis barrier.The number of SCs is directly proportional to the number of germ cells.And mature SCs ensure the growth of germ cells and the production of sperm.In this study,we explored the effect and underlying mechanism of Lycium barbarum polysaccharides(LBP)on primary SCs in young rats.Methods:Primary SCs were isolated from the testis of 20-day old rats.The cells were then treated with different concentrations of LBP.Immunocytochemistry was used to detect the expression of Ki67 and the androgen receptor(AR),and western blotting was used to detect the expression of cytokeratin-18(CK-18),AR and phosphorylated Akt(Ser473)in SCs.Results:The number of SCs increased significantly after LBP treatment,and the 100 mg/mL.LBP group had 14%more cells than the control group.The expression of Ki67 in LBP treated groups also increased significantly.LBP inhibited the expression of cytokeratin 18 in SCs.Besides,LBP increased the expression of AR on SCs and promoted the activation of Akt at the ser473 phosphorylation site.Conclusion:LBP promotes the proliferation of immature SCs in young rats and also accelerates their differentiation and maturation.This seems to be associated with activation of the Akt signaling pathway via up-regulation of AR.展开更多
基金Supported by National Natural Science Foundation of China,No.30871158 and No.81170604Outstanding Scientific Fund of Shengjing Hospital
文摘AIM:To investigate the change in intestinal dendritic cell(DC)number in fulminant hepatic failure(FHF).METHODS:An animal model of FHF was created.Intestinal CD11b/c was detected by immunohistochemistry and Western blot.Quantitative real-time polymerase chain reaction(PCR)was used to detect intestinal integrin-αm RNA expression.Intestinal CD83,CD86,CD74,CD3 and AKT were detected by immunohistochemistry,Western blot and PCR.Phosphorylated-AKT(p-AKT)was detected by immunohistochemistry and Western blot.RESULTS:In the FHF group[D-galactosamine(D-Galn)+lipopolysaccharide(LPS)group],the mice began to die after 6 h;conversely,in the D-Galn and LPS groups,the activity of mice was poor,but there were no deaths.Immunohistochemistry results showed that in FHF,the expression of CD11b/c(7988400±385941vs 1102400±132273,P<0.05),CD83(13875000±467493 vs 9257600±400364,P<0.05),CD86(7988400±385941 vs 1102400±13227,P<0.05)and CD74(11056000±431427 vs 4633400±267903,P<0.05)was significantly increased compared with the normal saline(NS)group.Compared with the NS group,the protein expression of CD11b/c(5.4817±0.77 vs 1.4073±0.37,P<0.05)and CD86(4.2673±0.69 vs 1.1379±0.42,P<0.05)was significantly increased.Itg-α(1.1224±0.3 vs 0.4907±0.19,P<0.05),CD83(3.6986±0.40 vs 1.0762±0.22,P<0.05)and CD86(1.5801±0.32 vs 0.8846±0.10,P<0.05)m RNA expression was increased significantly in the FHF group.At the protein level,expression of CD74in the FHF group(2.3513±0.52)was significantly increased compared with the NS group(1.1298±0.33),whereas in the LPS group(2.3891±0.47),the level of CD74 was the highest(P<0.05).At the gene level,the relative expression of CD74 m RNA in the FHF group(1.5383±0.26)was also significantly increased in comparison to the NS group(0.7648±0.22;P<0.05).CD3 expression was the highest in the FHF group(P<0.05).In the FHF,LPS and D-Galn groups,the expression of AKT at the protein and m RNA levels was elevated compared with the NS group,but there wasno statistical significance(P>0.05).The p-AKT protein expression in the FHF(1.54±0.06),LPS(1.56±0.05)and D-Galn(1.29±0.03)groups was higher than that in the NS group(1.07±0.03)(P<0.05).CONCLUSION:In FHF,a large number of DCs mature,express CD86,and activate MHC classⅡmolecular pathways to induce a T cell response,and the AKT pathway is activated.
基金We thank the National Natural Science Foundation of China(81273610)Research and Development Fund of Beijing University of Chinese Medicine(2019-ZFXZJJ-021)for financial support.
文摘Objective:Sertoli cells(SCs)provide physical support and material supply for germ cells and participate in the formation of blood-testis barrier.The number of SCs is directly proportional to the number of germ cells.And mature SCs ensure the growth of germ cells and the production of sperm.In this study,we explored the effect and underlying mechanism of Lycium barbarum polysaccharides(LBP)on primary SCs in young rats.Methods:Primary SCs were isolated from the testis of 20-day old rats.The cells were then treated with different concentrations of LBP.Immunocytochemistry was used to detect the expression of Ki67 and the androgen receptor(AR),and western blotting was used to detect the expression of cytokeratin-18(CK-18),AR and phosphorylated Akt(Ser473)in SCs.Results:The number of SCs increased significantly after LBP treatment,and the 100 mg/mL.LBP group had 14%more cells than the control group.The expression of Ki67 in LBP treated groups also increased significantly.LBP inhibited the expression of cytokeratin 18 in SCs.Besides,LBP increased the expression of AR on SCs and promoted the activation of Akt at the ser473 phosphorylation site.Conclusion:LBP promotes the proliferation of immature SCs in young rats and also accelerates their differentiation and maturation.This seems to be associated with activation of the Akt signaling pathway via up-regulation of AR.