AIM: To evaluate the antifibrotic effect of different doses of recombinant human Gamma-Interferon (IFN-gamma) in two rat models of hepatic fibrosis, and to observe its effect on moderate chronic hepatitis B virus fibr...AIM: To evaluate the antifibrotic effect of different doses of recombinant human Gamma-Interferon (IFN-gamma) in two rat models of hepatic fibrosis, and to observe its effect on moderate chronic hepatitis B virus fibrosis. METHODS: Hepatic fibrosis was successfully induced in 150 and 196 rats by subcutaneous injection of carbon tetrachloride (CCl4) and intraperitoneal injection of dimethylnitrosamine (DMN), respectively. Each of the two model groups was divided into: (1) fibrotic model group; (2) colchicine treatment group (0.1 mg/kg/day, gastrogavage for 8 weeks); (3) high-dose IFN-gamma group (15 MU/kg per day, i.m. for 8 weeks); (4) medium-dose IFN-gamma group (5 MU/kg daily, i.m. for 8 weeks); and (5) Y low-dose IFN-gamma group (1.67 MU/kg daily, i.m. for 8 weeks). Another group of 10 rats without any treatment was used as normal controls. At the end of the experiment, semi-quantitative histopathological scores of inflammation and fibrosis, liver alpha smooth muscle actin (alpha-SMA) expression level, liver hydroxyl proline content and serum hyaluronic acid levels were compared. And 47 medium chronic hepatitis B viral fibrosis patients were studied. They were given IFN-gamma treatment, 100 MU/day i.m. for the first three months and 100 MU qod i.m. for the next six months. Semi-quantitative pathological scores of inflammation and fibrosis and serum hepatic fibrosis indices were compared within the 9 months. RESULTS: In animal experiment, the pathological fibrosis scores and liver hydroxyl proline content were found to be significantly lower in rats treated with different doses of IFN-gamma as compared with rats in fibrotic model group induced by either CCl4 or DMN, in a dose-dependent manner. For CCl4-induced model, pathological fibrosis scores in high, medium and low doses IFN-gamma groups were 5.10 +/- 2.88, 7.70 +/- 3.53 and 8.00 +/- 3.30, respectively, but the score was 14.60 +/- 7.82 in fibrotic model group. Hydroxyl proline contents were 2.83 +/- 1.18, 3.59 +/- 1.22 and 4.80 +/- 1.62, in the three IFN-ga展开更多
Canine Interferon-γ(CaIFN-γ) cDNA was cloned from spleen T cells of dog by reverse transcription-polymerase chain reaction (RT-PCR). CaIFN-γ cDNA were digested with Hind Ⅲ and NotⅠ, and inserted into pRc/CMV2 exp...Canine Interferon-γ(CaIFN-γ) cDNA was cloned from spleen T cells of dog by reverse transcription-polymerase chain reaction (RT-PCR). CaIFN-γ cDNA were digested with Hind Ⅲ and NotⅠ, and inserted into pRc/CMV2 expression vector. The pRc/CMV2 /CaIFN-γ vector was sequenced, and predicted to produce a signal peptide of 23 amino acids and a mature protein of 143 amino acids with a molecular weight of 19 kD. Two potential N-glycosylation sites are located at positions 16 and 83 of the mature protein. Comparison of the CaIFN-γ protein sequence with that of CaIFN-γ reported from DDBJ/GenBank revealed a homology of 99%. To establish a long time expression system, pRc/CMV2/CaIFN-γ vector was transfected into mouse SP2/0 cell line. The SP2/0 cells culture supernatants was harvested and the antiviral activity was measured following cytopathic-effect inhibition assay using Madin-Darby Canine Kidney (MDCK)-vesicular stomatitis virus(VSV) system. Initial transformants with G418 phenotype produced recombinant CaIFN-γ titers ranging from 2,500 to 5,000 u/mL of culture medium.展开更多
目的研究干扰素(IFN)对肿瘤细胞表面Fas表达的调控,探讨其致凋亡机制。方法通过流式细胞仪测定肿瘤细胞的Fas表达,应用ELISA及Annexin V 染色检测细胞凋亡。对干扰素及抗Fas单克隆抗体诱导的肿瘤细胞凋亡进行评价。结果胃癌细胞系Kato-...目的研究干扰素(IFN)对肿瘤细胞表面Fas表达的调控,探讨其致凋亡机制。方法通过流式细胞仪测定肿瘤细胞的Fas表达,应用ELISA及Annexin V 染色检测细胞凋亡。对干扰素及抗Fas单克隆抗体诱导的肿瘤细胞凋亡进行评价。结果胃癌细胞系Kato-Ⅲ、AGS、肺癌细胞A549及喉癌细胞Hep-2均有不同程度的Fas表达,上述细胞与IFN-酌作用后Fas表达升高(P<0.05);IFN- 酌单独即可引起肿瘤细胞凋亡,同时可增加肿瘤细胞对抗Fas抗体的敏感性;IFN- 酌致凋亡及调节Fas表达的作用有时间依赖性,但不同细胞系对这两种因素致凋亡的敏感性不同。结论IFN治疗肿瘤的作用机制之一是增加肿瘤细胞Fas表达而致其凋亡,表明该药在肿瘤生物治疗中应用的合理性。展开更多
文摘AIM: To evaluate the antifibrotic effect of different doses of recombinant human Gamma-Interferon (IFN-gamma) in two rat models of hepatic fibrosis, and to observe its effect on moderate chronic hepatitis B virus fibrosis. METHODS: Hepatic fibrosis was successfully induced in 150 and 196 rats by subcutaneous injection of carbon tetrachloride (CCl4) and intraperitoneal injection of dimethylnitrosamine (DMN), respectively. Each of the two model groups was divided into: (1) fibrotic model group; (2) colchicine treatment group (0.1 mg/kg/day, gastrogavage for 8 weeks); (3) high-dose IFN-gamma group (15 MU/kg per day, i.m. for 8 weeks); (4) medium-dose IFN-gamma group (5 MU/kg daily, i.m. for 8 weeks); and (5) Y low-dose IFN-gamma group (1.67 MU/kg daily, i.m. for 8 weeks). Another group of 10 rats without any treatment was used as normal controls. At the end of the experiment, semi-quantitative histopathological scores of inflammation and fibrosis, liver alpha smooth muscle actin (alpha-SMA) expression level, liver hydroxyl proline content and serum hyaluronic acid levels were compared. And 47 medium chronic hepatitis B viral fibrosis patients were studied. They were given IFN-gamma treatment, 100 MU/day i.m. for the first three months and 100 MU qod i.m. for the next six months. Semi-quantitative pathological scores of inflammation and fibrosis and serum hepatic fibrosis indices were compared within the 9 months. RESULTS: In animal experiment, the pathological fibrosis scores and liver hydroxyl proline content were found to be significantly lower in rats treated with different doses of IFN-gamma as compared with rats in fibrotic model group induced by either CCl4 or DMN, in a dose-dependent manner. For CCl4-induced model, pathological fibrosis scores in high, medium and low doses IFN-gamma groups were 5.10 +/- 2.88, 7.70 +/- 3.53 and 8.00 +/- 3.30, respectively, but the score was 14.60 +/- 7.82 in fibrotic model group. Hydroxyl proline contents were 2.83 +/- 1.18, 3.59 +/- 1.22 and 4.80 +/- 1.62, in the three IFN-ga
文摘Canine Interferon-γ(CaIFN-γ) cDNA was cloned from spleen T cells of dog by reverse transcription-polymerase chain reaction (RT-PCR). CaIFN-γ cDNA were digested with Hind Ⅲ and NotⅠ, and inserted into pRc/CMV2 expression vector. The pRc/CMV2 /CaIFN-γ vector was sequenced, and predicted to produce a signal peptide of 23 amino acids and a mature protein of 143 amino acids with a molecular weight of 19 kD. Two potential N-glycosylation sites are located at positions 16 and 83 of the mature protein. Comparison of the CaIFN-γ protein sequence with that of CaIFN-γ reported from DDBJ/GenBank revealed a homology of 99%. To establish a long time expression system, pRc/CMV2/CaIFN-γ vector was transfected into mouse SP2/0 cell line. The SP2/0 cells culture supernatants was harvested and the antiviral activity was measured following cytopathic-effect inhibition assay using Madin-Darby Canine Kidney (MDCK)-vesicular stomatitis virus(VSV) system. Initial transformants with G418 phenotype produced recombinant CaIFN-γ titers ranging from 2,500 to 5,000 u/mL of culture medium.
文摘目的研究干扰素(IFN)对肿瘤细胞表面Fas表达的调控,探讨其致凋亡机制。方法通过流式细胞仪测定肿瘤细胞的Fas表达,应用ELISA及Annexin V 染色检测细胞凋亡。对干扰素及抗Fas单克隆抗体诱导的肿瘤细胞凋亡进行评价。结果胃癌细胞系Kato-Ⅲ、AGS、肺癌细胞A549及喉癌细胞Hep-2均有不同程度的Fas表达,上述细胞与IFN-酌作用后Fas表达升高(P<0.05);IFN- 酌单独即可引起肿瘤细胞凋亡,同时可增加肿瘤细胞对抗Fas抗体的敏感性;IFN- 酌致凋亡及调节Fas表达的作用有时间依赖性,但不同细胞系对这两种因素致凋亡的敏感性不同。结论IFN治疗肿瘤的作用机制之一是增加肿瘤细胞Fas表达而致其凋亡,表明该药在肿瘤生物治疗中应用的合理性。
