AIM: To investigate if ischemia/reperfusion (I/R) injury in aged liver could be alleviated by heme oxygenase-1 (HO-1).METHODS: Three groups of SD rats (16 mo old) were studied. Group 1: control donors received physiol...AIM: To investigate if ischemia/reperfusion (I/R) injury in aged liver could be alleviated by heme oxygenase-1 (HO-1).METHODS: Three groups of SD rats (16 mo old) were studied. Group 1: control donors received physiological saline 24 h before their livers were harvested; group 2: donors were pretreated with hemih 24 h before their livers were harvested; and group 3: donors received hemin 24 h before their livers were harvested and zinc protoporphyrin (ZnPP,HO-1 inhibitor) was given to recipients at reperfusion. The harvested livers were stored in University of Wisconsin solution (4 ℃) for 6 h, and then transplanted to syngeneic rats. Serum glutamic oxaloacetic transaminase (SGOT),apoptotic cells, and apoptotic gene were measured 3, 6,12, 24, 48 h after reperfusion. We measured the apoptotic index by TUNEL, determined the expression of antiapoptotic Bcl-2 and proapoptotic (caspase-3) gene products by Western blot.RESULTS: After 3, 6, 12, 24, and 48 h of reperfusion, the SGOT levels (584.4±85.8 u/L, 999.2±125.2 u/L, 423.4±161.3u/L, 257.8±95.8 u/L, and 122.4±26.4 u/L) in hemin group were significantly (all P＜0.05) lower than those in saline group (1082.2±101.2 u/L, 1775.2±328.3 u/L, 840.4±137.8 u/L,448.6±74.3 u/L, and 306.2±49.3 u/L). Liver HO-1 enzymatic activity correlated with beneficial effects of hemin and deleterious effects of adjunctive ZnPP treatment. Markedly less apoptotic (TUNEL+) liver cells 3, 6, 12, 24, and 48 h after reperfusion (5.16±0.73, 10.2±0.67, 9.28±0.78, 7.14±1.12,and 4.78±0.65) (P＜0.05) could be detected in hemin liver grafts, as compared to controls (7.82±1.05, 15.94±1.82,11.67±1.59, 8.28±1.09, and 6.36±0.67). We detected the increased levels of Bcl-2 (1.5-fold) expression and compared with saline controls. These differences were most pronounced at 12 h after transplantation. In contrast, an active form of proapoptotic caspase-3 (p20) protein was found to be 2.9-fold lower at 24 h in hemin-pretreated group, as compared to saline liver transplant controls.CONCLUSION: HO-1 overexpression can provide potent protection against cold I/R injury. This effect depends, at least in part, on HO-1-mediated inhibition of antiapoptotic mechanism.展开更多
Direct intratumoral introduction of therapeutic or regulatory genes is a developing technology with potential application for cancer gene therapy.Macrophage inflammatory protein-I beta(MIP-Iβ)is a chemokine which can...Direct intratumoral introduction of therapeutic or regulatory genes is a developing technology with potential application for cancer gene therapy.Macrophage inflammatory protein-I beta(MIP-Iβ)is a chemokine which can chemoattract immune cells such as T cells.In the present study,murine colorectal adenocarcinoma CT26 cells were transfected with a recombinant adenovirus (AdhMIP-Iβ)carrying the human MIP-Iβ gene.24 h post-transfection,hMIP-1β levels reached approximately 980 pg/ml in supernatants of 10~6 hMIP-Iβ-transfected CT26 cells.Moreover,the supernatants exhibited chemotactic activity for CD8^+ T cells,CD4^+ T cells,NK cells and immature DCs.Intratumoral injection of AdhMIP-Iβ significantly inhibited tumor growth and prolonged the survival time of tumor-bearing mice.Intratumoral hMIP-Iβ gene transfer also induced powerful tumor-specific CTL responses in vivo.The therapeutic effects of hMIP-Iβ gene therapy were greatly reduced following in vivo depletion of both CD4^+ and CD8^+ cells,but were unaffected by depletion of single T cell subsets.Immune cell depletion experiments also revealed that NK cells played an important role in hMIP-1β-induced antitumor responses.These results suggest that intratumoral expression of hMIP-1β has the potential effect to induce host antitumor immunity and may prove to be a useful form of cancer gene therapy. Cellular & Molecular Immunology.2004;1(3):199-204.展开更多
基金Supported by the "135" Medical Project of Jiangsu, No. 135-10
文摘AIM: To investigate if ischemia/reperfusion (I/R) injury in aged liver could be alleviated by heme oxygenase-1 (HO-1).METHODS: Three groups of SD rats (16 mo old) were studied. Group 1: control donors received physiological saline 24 h before their livers were harvested; group 2: donors were pretreated with hemih 24 h before their livers were harvested; and group 3: donors received hemin 24 h before their livers were harvested and zinc protoporphyrin (ZnPP,HO-1 inhibitor) was given to recipients at reperfusion. The harvested livers were stored in University of Wisconsin solution (4 ℃) for 6 h, and then transplanted to syngeneic rats. Serum glutamic oxaloacetic transaminase (SGOT),apoptotic cells, and apoptotic gene were measured 3, 6,12, 24, 48 h after reperfusion. We measured the apoptotic index by TUNEL, determined the expression of antiapoptotic Bcl-2 and proapoptotic (caspase-3) gene products by Western blot.RESULTS: After 3, 6, 12, 24, and 48 h of reperfusion, the SGOT levels (584.4±85.8 u/L, 999.2±125.2 u/L, 423.4±161.3u/L, 257.8±95.8 u/L, and 122.4±26.4 u/L) in hemin group were significantly (all P＜0.05) lower than those in saline group (1082.2±101.2 u/L, 1775.2±328.3 u/L, 840.4±137.8 u/L,448.6±74.3 u/L, and 306.2±49.3 u/L). Liver HO-1 enzymatic activity correlated with beneficial effects of hemin and deleterious effects of adjunctive ZnPP treatment. Markedly less apoptotic (TUNEL+) liver cells 3, 6, 12, 24, and 48 h after reperfusion (5.16±0.73, 10.2±0.67, 9.28±0.78, 7.14±1.12,and 4.78±0.65) (P＜0.05) could be detected in hemin liver grafts, as compared to controls (7.82±1.05, 15.94±1.82,11.67±1.59, 8.28±1.09, and 6.36±0.67). We detected the increased levels of Bcl-2 (1.5-fold) expression and compared with saline controls. These differences were most pronounced at 12 h after transplantation. In contrast, an active form of proapoptotic caspase-3 (p20) protein was found to be 2.9-fold lower at 24 h in hemin-pretreated group, as compared to saline liver transplant controls.CONCLUSION: HO-1 overexpression can provide potent protection against cold I/R injury. This effect depends, at least in part, on HO-1-mediated inhibition of antiapoptotic mechanism.
基金supported by Grants from National Natural Science Foundation of China(30271202)Natural High Technology Research and Development Program of China(2003AA215040)the National Key Basic Research Program of China(2001CB510002).
文摘Direct intratumoral introduction of therapeutic or regulatory genes is a developing technology with potential application for cancer gene therapy.Macrophage inflammatory protein-I beta(MIP-Iβ)is a chemokine which can chemoattract immune cells such as T cells.In the present study,murine colorectal adenocarcinoma CT26 cells were transfected with a recombinant adenovirus (AdhMIP-Iβ)carrying the human MIP-Iβ gene.24 h post-transfection,hMIP-1β levels reached approximately 980 pg/ml in supernatants of 10~6 hMIP-Iβ-transfected CT26 cells.Moreover,the supernatants exhibited chemotactic activity for CD8^+ T cells,CD4^+ T cells,NK cells and immature DCs.Intratumoral injection of AdhMIP-Iβ significantly inhibited tumor growth and prolonged the survival time of tumor-bearing mice.Intratumoral hMIP-Iβ gene transfer also induced powerful tumor-specific CTL responses in vivo.The therapeutic effects of hMIP-Iβ gene therapy were greatly reduced following in vivo depletion of both CD4^+ and CD8^+ cells,but were unaffected by depletion of single T cell subsets.Immune cell depletion experiments also revealed that NK cells played an important role in hMIP-1β-induced antitumor responses.These results suggest that intratumoral expression of hMIP-1β has the potential effect to induce host antitumor immunity and may prove to be a useful form of cancer gene therapy. Cellular & Molecular Immunology.2004;1(3):199-204.
