CAJ | 학술논문

目的：观察辛伐他汀和氟伐他汀对大鼠胰岛β细胞胰岛素合成的抑制作用及机制。方法8周龄健康雄性Wistar大鼠90只，体重250～300 g，饲养1周内处死。采用胆管注射胶原酶法提取大鼠胰岛，分为对照组（11.1 mmol/L RPMI 1640培养液）、辛伐他汀组（100μmol/L辛伐他汀+11.1 mmol/L RPMI 1640培养液）和氟伐他汀组（100μmol/L氟伐他汀+11.1 mmol/L RPMI 1640培养液）。辛伐他汀、氟伐他汀作用24 h后，放射免疫法测定大鼠胰岛β细胞胰岛素含量的变化，荧光定量聚合酶链反应（PCR）分析大鼠胰岛β细胞胰岛素mRNA的表达；构建人胰岛素启动子-荧光素酶质粒，脂质体转染MIN6细胞，双荧光素酶法观察辛伐他汀、氟伐他汀作用24 h、48 h后对胰岛素启动子活性的影响。三组间数据比较采用方差分析。结果与对照组相比,氟伐他汀和辛伐他汀作用24 h,均使胰岛β细胞胰岛素含量明显减少,分别较对照组减少21.1%和27.1%（q′=5.08和6.54，P<0.05）。氟伐他汀和辛伐他汀均明显抑制胰岛素mRNA的表达，作用前后分别为（0.155±0.013比0.265±0.030和0.153±0.031比0.265±0.030，q′=9.488、9.661，P<0.05）。与对照组相比，辛伐他汀作用24 h后，荧光素酶活性受到明显抑制（1.11±0.26比1.29±0.39，q′=3.55，P<0.05),氟伐他汀作用48 h后明显抑制荧光素酶活性（1.14±0.29比1.29±0.35，q′=5.76，P<0.01）。结论辛伐他汀、氟伐他汀通过抑制大鼠胰岛β细胞胰岛素mRNA表达而抑制胰岛素合成，这种抑制效应可能与其对胰岛素启动子活性的抑制有关。
목적：관찰신벌타정화불벌타정대대서이도β세포이도소합성적억제작용급궤제。방법8주령건강웅성Wistar대서90지，체중250～300 g，사양1주내처사。채용담관주사효원매법제취대서이도，분위대조조（11.1 mmol/L RPMI 1640배양액）、신벌타정조（100μmol/L신벌타정+11.1 mmol/L RPMI 1640배양액）화불벌타정조（100μmol/L불벌타정+11.1 mmol/L RPMI 1640배양액）。신벌타정、불벌타정작용24 h후，방사면역법측정대서이도β세포이도소함량적변화，형광정량취합매련반응（PCR）분석대서이도β세포이도소mRNA적표체；구건인이도소계동자-형광소매질립，지질체전염MIN6세포，쌍형광소매법관찰신벌타정、불벌타정작용24 h、48 h후대이도소계동자활성적영향。삼조간수거비교채용방차분석。결과여대조조상비,불벌타정화신벌타정작용24 h,균사이도β세포이도소함량명현감소,분별교대조조감소21.1%화27.1%（q′=5.08화6.54，P<0.05）。불벌타정화신벌타정균명현억제이도소mRNA적표체，작용전후분별위（0.155±0.013비0.265±0.030화0.153±0.031비0.265±0.030，q′=9.488、9.661，P<0.05）。여대조조상비，신벌타정작용24 h후，형광소매활성수도명현억제（1.11±0.26비1.29±0.39，q′=3.55，P<0.05),불벌타정작용48 h후명현억제형광소매활성（1.14±0.29비1.29±0.35，q′=5.76，P<0.01）。결론신벌타정、불벌타정통과억제대서이도β세포이도소mRNA표체이억제이도소합성，저충억제효응가능여기대이도소계동자활성적억제유관。
Objective To evaluate the inhibitory effects and the mechanisms of simvastatin and fluvastatin on insulin synthesis from pancreatic islets in rats. Methods The pancreatic islets were isolated from ninety eight-week-old male Wistar rats(body weight 250-300 g) by injecting collagen through the bile duct. The isolated pancreatic islets were divided into control group(treated with 11.1 mmol/L RPMI 1640), simvastatin group and fluvastatin group. Insulin content in pancreatic islets from rats was measured by radioimmunoassay(RIA), and the mRNA expression was accessed by real-time polymerase chain reaction after treatment for 24 hours with simvastatin or fluvastatin(100 μmol/L). The human insulin promoter-luciferase vector was constructed and MIN6 cells were transfected by the vector. The activity of insulin promoter was measured by the dual-luciferase reporter assay system after the treatment by simvastatin and fluvastatin for 24 and 48 hours respectively. One-way analysis of variance was carried out to analyze all the data. Results Compared with that in control group, the RIA results showed the insulin content was significantly decreased after the treatment of simvastatin and fulvastatin for 24 hours by 21.1%and 27.1%respectively(q′=5.08 and 6.54, P<0.05). Fulvastatin and simvastatin significantly inhibited the expression of insulin mRNA, the insulin mRNA were significantly decreased in both fulvastatin group and simvastatin group(0.155±0.013 vs 0.265±0.030, 0.153±0.031 vs 0.265±0.030,q′=9.488, 9.661, P<0.05). Compared with that in control group, the activity of luciferase was significantly inhibited by simvastatin treatment for 24 hours (1.11±0.26 vs 1.29±0.39, q′=3.55,P<0.05) and fluvastatin treatment for 48 hours (1.14±0.29 vs 1.29± 0.35, q′=5.76,P<0.01). Conclusion Simvastatin and fluvastatin inhibite the insulin synthesis by inhibiting the expression of insulin mRNA in rat pancreatic islet βcells, which may be related to its inhibitory effect on the activity of insulin promoter.