Objective: To discuss the relationship between hypercholesterolemic disease and the functional and structural changes of Sphincter of Oddi (SO) by the study of effect of Cholesterol Liposome (CL) on structural and qua...Objective: To discuss the relationship between hypercholesterolemic disease and the functional and structural changes of Sphincter of Oddi (SO) by the study of effect of Cholesterol Liposome (CL) on structural and quantitative changes of SO cells. Methods: Rabbit SO was isolated for primary cell culture and subculture. After subcultured with different concentration of CL culture medium for 20 h, the structural and quantitative changes of SO cells were analyzed and detected by MTT-test, flow cytometer (FCM), electronic microscope and electrophoresis technique respectively. Results: CL contributed a prominent stimulus to SO cells proliferation at middle concentration (<0. 5 - 0. 8 mg/ml), which could be confirmed by FCM analysis which indicated the number of SO cells in S-phase increasing remarkably; however, high concentration of CL inhibited SO cells' proliferation (>1. 0 mg/ml) and induced apoptosis of SO cells. Swelled mitochondria and dilated endoplasmic reticulum as well as disjoined and diminished microfilaments were found in SO cells by electronic microscopy. The content of SO cells actin decreased with the increment of cholesterol concentration. There was a significant difference of actin content between CL groups and control group (P<0. 05). Conclusion: CL may change SO cell membrane's function, organelle's structure and especially the quantity and configuration of microfilaments, at the same time, CL at different concentration can induce changes of SO cells cycle and lead to different changes in the number of SO cells.展开更多
目的研究兔胆道口括约肌(sphincter of Oddi,SO)平滑肌细胞在胆固醇脂质体作用后收缩性的变化,探讨高胆固醇血症兔SO 动力异常的机制方法取纯种新西兰兔 SO 段,用Ⅱ型胶原酶消化获得单个平滑肌细胞,与胆崮醇/卵磷脂摩尔比为2:1和0.5:1...目的研究兔胆道口括约肌(sphincter of Oddi,SO)平滑肌细胞在胆固醇脂质体作用后收缩性的变化,探讨高胆固醇血症兔SO 动力异常的机制方法取纯种新西兰兔 SO 段,用Ⅱ型胶原酶消化获得单个平滑肌细胞,与胆崮醇/卵磷脂摩尔比为2:1和0.5:1的胆固醇脂质体(1g/L)分别孵育2 h 后,用不同浓度 KCl(9-24)nmol/L,乙酰胆碱(10^(12)～10^(-6))mol/L 作用于平滑肌细胞,激动剂作用后30s,加入丙烯醛固定,分别测量各组细胞的收缩百分比.结果兔 SO 平滑肌细胞平均长度为(143.7±12.3)μm,经胆固醇脂质体作用后平均长度无明显变化,对 KCl 和乙酰胆碱呈浓度依赖性收缩.KCl(18mmol/L)诱导最大收缩比为22.2%±0.7%,而2:1的胆固醇脂质体作用后收缩比为16.5%±0.6%(P<0.01);乙酰胆碱(10^(-7)mol/L)诱导最大收缩比为20.3%±1.4%,2:1的胆固醇脂质体作用后收缩比为16.5%±1.3%(P<0.05).摩尔比为0.5:1胆固醇脂质体作用后最大收缩比分别为21.3%±1.4%和19.2%±1.1%,同对照组相比无显著下降.结论兔 SO 平滑肌细胞经摩尔比2:1的胆固醇脂质体后起收缩性下降,推测这是高胆固醇血症可以导致兔 SO 的动力异常的机制.展开更多
文摘Objective: To discuss the relationship between hypercholesterolemic disease and the functional and structural changes of Sphincter of Oddi (SO) by the study of effect of Cholesterol Liposome (CL) on structural and quantitative changes of SO cells. Methods: Rabbit SO was isolated for primary cell culture and subculture. After subcultured with different concentration of CL culture medium for 20 h, the structural and quantitative changes of SO cells were analyzed and detected by MTT-test, flow cytometer (FCM), electronic microscope and electrophoresis technique respectively. Results: CL contributed a prominent stimulus to SO cells proliferation at middle concentration (<0. 5 - 0. 8 mg/ml), which could be confirmed by FCM analysis which indicated the number of SO cells in S-phase increasing remarkably; however, high concentration of CL inhibited SO cells' proliferation (>1. 0 mg/ml) and induced apoptosis of SO cells. Swelled mitochondria and dilated endoplasmic reticulum as well as disjoined and diminished microfilaments were found in SO cells by electronic microscopy. The content of SO cells actin decreased with the increment of cholesterol concentration. There was a significant difference of actin content between CL groups and control group (P<0. 05). Conclusion: CL may change SO cell membrane's function, organelle's structure and especially the quantity and configuration of microfilaments, at the same time, CL at different concentration can induce changes of SO cells cycle and lead to different changes in the number of SO cells.
文摘目的研究兔胆道口括约肌(sphincter of Oddi,SO)平滑肌细胞在胆固醇脂质体作用后收缩性的变化,探讨高胆固醇血症兔SO 动力异常的机制方法取纯种新西兰兔 SO 段,用Ⅱ型胶原酶消化获得单个平滑肌细胞,与胆崮醇/卵磷脂摩尔比为2:1和0.5:1的胆固醇脂质体(1g/L)分别孵育2 h 后,用不同浓度 KCl(9-24)nmol/L,乙酰胆碱(10^(12)～10^(-6))mol/L 作用于平滑肌细胞,激动剂作用后30s,加入丙烯醛固定,分别测量各组细胞的收缩百分比.结果兔 SO 平滑肌细胞平均长度为(143.7±12.3)μm,经胆固醇脂质体作用后平均长度无明显变化,对 KCl 和乙酰胆碱呈浓度依赖性收缩.KCl(18mmol/L)诱导最大收缩比为22.2%±0.7%,而2:1的胆固醇脂质体作用后收缩比为16.5%±0.6%(P<0.01);乙酰胆碱(10^(-7)mol/L)诱导最大收缩比为20.3%±1.4%,2:1的胆固醇脂质体作用后收缩比为16.5%±1.3%(P<0.05).摩尔比为0.5:1胆固醇脂质体作用后最大收缩比分别为21.3%±1.4%和19.2%±1.1%,同对照组相比无显著下降.结论兔 SO 平滑肌细胞经摩尔比2:1的胆固醇脂质体后起收缩性下降,推测这是高胆固醇血症可以导致兔 SO 的动力异常的机制.
