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Transplant renal arteriopathy in humans is associated with production of fibronectin and the induction of migrating smooth muscle cells
Tuesday 11 October 1994: Poster Abstracts Arterial wall matrix
94
El
Lisinopril, an ACE inhibitor, suppresses atherogenesis in the aorta but not the coronary arteries of cholesterol-fed rabbits NT, Fujinishi A, Yashiro A, Takahara K, Nakashima Y, Kuroiwa A, 2nd Dept. of Int. Med., Univ. of Occupational and Environmental Health, Sch. of Med., 1-I Iseigaoka Yahata, Nishiku, Kitakyushu, Japan 807
The effect of ACE inhibitors on atherogenesis remains controversial. In this study, to assess the effect of Lisinopril on atherosclerotic change of aorta and coronary artery induced by hypercholesterolemia, 1% cholesterol diet rabbits were treated with Lisinopril (3 mg or 6 mg/day, intraperitoneal injection) or saline. Eight were fed standard diet and treated with saline as control. At the end of 10 weeks, aortas obtained after sacrifice were used for the measurements of macroscopic atherosclerotic area and biochemical analysis of aortic wall. The effect of Lisinopril on coronary atherosclerosis was also evaluated histologically. The aortic content of cholesterol of the rabbits with Lisinopril was significantly less than those without Lisinopril [29.9 f 5.4 (saline) vs 13.3 f 2.6 (Lisnopril 3 mg), 14.Ort 3.4 (Lisinopril 6 mg): mean f SE]. The percent involvement of aortic surface in the rabbits treated with Lisinopril was significantly lower than that without Lisinopril [18.7 + 2.6 (saline) vs 9.8 f 3.9 (Lisinopril 3 mg), 5.1 f 1.3 (Lisinopril 6 mg)]. There, however, were no differences in histological findings in arteries between the rabbits with or without Lisinopril treatment. These results suggest that Lisinopril might suppress atherogenesis in the aorta but not in the coronary artery of cholesterolfed rabbits.
WY.
We have shown recently that matrix metalloproteinases (MMPs) are produced by aortic SMC in response to PDGF. For a better understanding of the mechanism of atherogenesis, in vivo or in vitro synthesis of MMPs by SMC was analyzed by immunohistochemistry and immunoblotting. Elastase activity in the culture medium was also estimated. Production of proMMP 1, 2, and 3 was detected in cultured SMC isolated from the aortas of both neonates and fetuses; in medial SMC cultured from young individuals, production of proMMP-I and -3 was extremely decreased, but was apparent in intimal SMC. Immunohistochemical observation indicated that in the media of fetal or neonatal aorta, SMC synthesized large amounts of the three proMMPs; in aortas from older individuals, proMMP-2, but not proMMP-1 and -3, was detected in the media, and relatively large amounts of proMMP-1, -2, and -3 were produced by SMC in the slightly thickened intima. Assay of elastase activity in the culture medium gave results similar to those for MMPs. We conclude that the production of proMMP-1 and -3 is associated with phenotypic modulation of SMC to a ‘synthetic’ state, and that the ability of SMC to produce MMPs plays an important role in the development and/or aging of the human aorta through remodeling of the extracellular matrix; furthermore elastase is also involved in these processes in the arterial wall. 181
Transplant renal arteriopathy in humans is associated with production of fibronectin and the induction of migrating smooth muscle cells mS, Ueda M, Han Y-S, Nakatani T, Kishimoto T, Suzuki S, Amemiya H, Dept. of Urology, Osaka City Univ. Med. Sch., l-5161
Murahashi N*, Kate S*, Hiraoka K*, Morimatsu M*,
Dept. of Pathol., Univ. of Occupational and Enviromental Health, Japan, Sch. of Med., Yahata Nishi-ku, Kitakyushu 807: *Dept. of Pathol., Kurume Univ. Sch. of Med., Kurume 830, Japan
Immortalization of human coronary endothelial cells - a comparison between immortalized coronary and aortic endothelial cell lines Murahashi N, Tanimoto A, Arima N, Hamada T, Sasa-
wR. guri Y, Dept. of Pathol., Univ. of Occupational and Environmental Health, Sch. of Med., Japan
Yahatanishiku, Kitakyushu 807,
7, Asahi-machi, Abeno-ku, Osaka, Japan
Transplant renal arteriopathy (TRA) is one of the major obstacles to long-term survival of human renal allografts. However, the pathobiologic mechanisms of human TRA are poorly understood. In this study, we immunohistochemically investigated expression of fibronectin in association with cellular components of TRA and with phenotypic expqession of smooth muscle cells. Ten renal allogrifts that had been removed due to rejection were available for tlus study. Time lapse between transplantation and removal of the renal allografts in these patients ranged from 1 month to 4 years. Monoclonal antibodies used were as follows: anti-T cell antibody, UCHLl; anti-B cell antibody, HAM56; antimuscle actin antibody, HHF35; anti-smooth muscle actin antibody, CGA7; anti-vimentin antibody; and anti-fibronectin (FN) antibody. In young proliferative lesions of TRA (l-3 months after transplantation), enhanced expression of FN was observed in the media and neointima. In these lesions, the neointima consisted largely of macrophages and T cells, intermixed with smooth muscle cells of de-differentiated phenotype. In older lesions of TRA (8 months to 4 years after transplantation), no or only weak expression of FN was observed in the neointima. At these stages, the neointima consisted almost entirely of smooth muscle cells of differentiated phenotype. Arteries in the normal renal tissues (n = 7) examined as controls showed no expression of FN. These results suggest that the pathophysiology of TRA involves an inflammatory process associated with production of fibronectin and the induction of migrating smooth muscle cells. 171
Development-related changes in matrix metalloproteinase expression in human aortic smooth muscle cells
Endothelial cells have many physiological functions such as regulation of vascular permeability, prevention of coagulation and platelet deposition, regulation of the vascular tone, and angiogenesis. Therefore, it is important to investigate these functions of the cells under physiological or pathological conditions in vi&o. It is known, however, that functions of endothelial cells differ somewhat from cell to cell depending on their site of origin. Since it is difficult to subculture human endothelial cells for several passages, we established an immortalized endothelial cell line from human coronary artery and compared its functions with those of an aortic endothelial cell line which had been reported previously. The primary coronary endothelial cells were isolated from an autopsy case of a 76-year-old man. The cells were immortalized with origin-minus SV40 plasmid by the liposome transfection method. One colony was isolated from colonies in the transfected primary cultures 2 weeks after the transfection. The cells expressed SV40 large T antigen in the nuclei, Factor VIII-related antigen and CJlex europaeus I agglutinin in the cytoplasm. The synthesis of prostacyclin was detected in both cell lines. Coronary endothelial cells were smaller than aortic endothelial cells. Assay of plasminogen activator revealed that large amounts of the enzyme were produced by the coronary cell line but lesser amounts by the aortic cell line. Western blotting showed the cells to produce matrix metalloproteinases, which were activated by plasminogen. We conclude that these cells lines will be useful for investigation of endotbelial cell functions. 191 W,
Gemtibrozll suppresses atherosclerosis by preventing foam cell infiltration Saku K, Zhang B, Liu R, Bai H, Hirata K, Takebayashi S,
Atherosclerosis X, Montreal, October I994
94
El
Lisinopril, an ACE inhibitor, suppresses atherogenesis in the aorta but not the coronary arteries of cholesterol-fed rabbits NT, Fujinishi A, Yashiro A, Takahara K, Nakashima Y, Kuroiwa A, 2nd Dept. of Int. Med., Univ. of Occupational and Environmental Health, Sch. of Med., 1-I Iseigaoka Yahata, Nishiku, Kitakyushu, Japan 807
The effect of ACE inhibitors on atherogenesis remains controversial. In this study, to assess the effect of Lisinopril on atherosclerotic change of aorta and coronary artery induced by hypercholesterolemia, 1% cholesterol diet rabbits were treated with Lisinopril (3 mg or 6 mg/day, intraperitoneal injection) or saline. Eight were fed standard diet and treated with saline as control. At the end of 10 weeks, aortas obtained after sacrifice were used for the measurements of macroscopic atherosclerotic area and biochemical analysis of aortic wall. The effect of Lisinopril on coronary atherosclerosis was also evaluated histologically. The aortic content of cholesterol of the rabbits with Lisinopril was significantly less than those without Lisinopril [29.9 f 5.4 (saline) vs 13.3 f 2.6 (Lisnopril 3 mg), 14.Ort 3.4 (Lisinopril 6 mg): mean f SE]. The percent involvement of aortic surface in the rabbits treated with Lisinopril was significantly lower than that without Lisinopril [18.7 + 2.6 (saline) vs 9.8 f 3.9 (Lisinopril 3 mg), 5.1 f 1.3 (Lisinopril 6 mg)]. There, however, were no differences in histological findings in arteries between the rabbits with or without Lisinopril treatment. These results suggest that Lisinopril might suppress atherogenesis in the aorta but not in the coronary artery of cholesterolfed rabbits.
WY.
We have shown recently that matrix metalloproteinases (MMPs) are produced by aortic SMC in response to PDGF. For a better understanding of the mechanism of atherogenesis, in vivo or in vitro synthesis of MMPs by SMC was analyzed by immunohistochemistry and immunoblotting. Elastase activity in the culture medium was also estimated. Production of proMMP 1, 2, and 3 was detected in cultured SMC isolated from the aortas of both neonates and fetuses; in medial SMC cultured from young individuals, production of proMMP-I and -3 was extremely decreased, but was apparent in intimal SMC. Immunohistochemical observation indicated that in the media of fetal or neonatal aorta, SMC synthesized large amounts of the three proMMPs; in aortas from older individuals, proMMP-2, but not proMMP-1 and -3, was detected in the media, and relatively large amounts of proMMP-1, -2, and -3 were produced by SMC in the slightly thickened intima. Assay of elastase activity in the culture medium gave results similar to those for MMPs. We conclude that the production of proMMP-1 and -3 is associated with phenotypic modulation of SMC to a ‘synthetic’ state, and that the ability of SMC to produce MMPs plays an important role in the development and/or aging of the human aorta through remodeling of the extracellular matrix; furthermore elastase is also involved in these processes in the arterial wall. 181
Transplant renal arteriopathy in humans is associated with production of fibronectin and the induction of migrating smooth muscle cells mS, Ueda M, Han Y-S, Nakatani T, Kishimoto T, Suzuki S, Amemiya H, Dept. of Urology, Osaka City Univ. Med. Sch., l-5161
Murahashi N*, Kate S*, Hiraoka K*, Morimatsu M*,
Dept. of Pathol., Univ. of Occupational and Enviromental Health, Japan, Sch. of Med., Yahata Nishi-ku, Kitakyushu 807: *Dept. of Pathol., Kurume Univ. Sch. of Med., Kurume 830, Japan
Immortalization of human coronary endothelial cells - a comparison between immortalized coronary and aortic endothelial cell lines Murahashi N, Tanimoto A, Arima N, Hamada T, Sasa-
wR. guri Y, Dept. of Pathol., Univ. of Occupational and Environmental Health, Sch. of Med., Japan
Yahatanishiku, Kitakyushu 807,
7, Asahi-machi, Abeno-ku, Osaka, Japan
Transplant renal arteriopathy (TRA) is one of the major obstacles to long-term survival of human renal allografts. However, the pathobiologic mechanisms of human TRA are poorly understood. In this study, we immunohistochemically investigated expression of fibronectin in association with cellular components of TRA and with phenotypic expqession of smooth muscle cells. Ten renal allogrifts that had been removed due to rejection were available for tlus study. Time lapse between transplantation and removal of the renal allografts in these patients ranged from 1 month to 4 years. Monoclonal antibodies used were as follows: anti-T cell antibody, UCHLl; anti-B cell antibody, HAM56; antimuscle actin antibody, HHF35; anti-smooth muscle actin antibody, CGA7; anti-vimentin antibody; and anti-fibronectin (FN) antibody. In young proliferative lesions of TRA (l-3 months after transplantation), enhanced expression of FN was observed in the media and neointima. In these lesions, the neointima consisted largely of macrophages and T cells, intermixed with smooth muscle cells of de-differentiated phenotype. In older lesions of TRA (8 months to 4 years after transplantation), no or only weak expression of FN was observed in the neointima. At these stages, the neointima consisted almost entirely of smooth muscle cells of differentiated phenotype. Arteries in the normal renal tissues (n = 7) examined as controls showed no expression of FN. These results suggest that the pathophysiology of TRA involves an inflammatory process associated with production of fibronectin and the induction of migrating smooth muscle cells. 171
Development-related changes in matrix metalloproteinase expression in human aortic smooth muscle cells
Endothelial cells have many physiological functions such as regulation of vascular permeability, prevention of coagulation and platelet deposition, regulation of the vascular tone, and angiogenesis. Therefore, it is important to investigate these functions of the cells under physiological or pathological conditions in vi&o. It is known, however, that functions of endothelial cells differ somewhat from cell to cell depending on their site of origin. Since it is difficult to subculture human endothelial cells for several passages, we established an immortalized endothelial cell line from human coronary artery and compared its functions with those of an aortic endothelial cell line which had been reported previously. The primary coronary endothelial cells were isolated from an autopsy case of a 76-year-old man. The cells were immortalized with origin-minus SV40 plasmid by the liposome transfection method. One colony was isolated from colonies in the transfected primary cultures 2 weeks after the transfection. The cells expressed SV40 large T antigen in the nuclei, Factor VIII-related antigen and CJlex europaeus I agglutinin in the cytoplasm. The synthesis of prostacyclin was detected in both cell lines. Coronary endothelial cells were smaller than aortic endothelial cells. Assay of plasminogen activator revealed that large amounts of the enzyme were produced by the coronary cell line but lesser amounts by the aortic cell line. Western blotting showed the cells to produce matrix metalloproteinases, which were activated by plasminogen. We conclude that these cells lines will be useful for investigation of endotbelial cell functions. 191 W,
Gemtibrozll suppresses atherosclerosis by preventing foam cell infiltration Saku K, Zhang B, Liu R, Bai H, Hirata K, Takebayashi S,
Atherosclerosis X, Montreal, October I994