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钩藤碱固体脂质纳米粒对TGF-β1诱导大鼠血管平滑肌细胞增殖的影响

Inhibitory effects of solid lipid nanoparticles of rhynchophylline on proliferation of rat vasculars mooth muscle cells induced by TGF-β1

摘要目的 研究钩藤碱固体脂质纳米粒对TGF-β1诱导的大鼠血管平滑肌细胞增殖的影响,并探讨其作用机制.方法 采用组织块培养法进行大鼠胸主动脉血管平滑肌细胞的原代培养,将细胞按随机数字表法分为对照组,TGF-β1组,TGF-β1+钩藤碱固体脂质纳米粒高、中、低剂量组.除对照组外,其余各组细胞加入20 μg/L的TGF-β1进行干预,TGF-β1+钩藤碱固体脂质纳米粒高、中、低剂量组细胞加入25、50、100 mg/L的钩藤碱固体脂质纳米粒进行干预.培养24 h后,采用MTT法检测各组细胞增殖抑制率,采用流式细胞仪检测各组细胞周期,采用Western blot法检测各组细胞c-myc、c-Fos蛋白表达.结果 与 TGF-β1组比较,TGF-β1+钩藤碱固体脂质纳米粒高剂量组吸光度值[(0.457±0.046)比(0.975±0.049)]降低(P<0.01);TGF-β1+钩藤碱固体脂质纳米粒高、中、低剂量组 S 期细胞[(15.87± 2.47)%、(15.23±1.69)%、(17.02±2.87)%比(38.58±2.68)%]比例降低(P<0.01),c-myc[(48.65±3.65)、(50.69±4.16)、(55.29±3.67)比(68.21±3.25)]、c-Fos[(38.78±4.25)、(43.56±3.69)、(46.58±3.57)比(66.54±4.09)]蛋白表达降低(P<0.01).结论 钩藤碱固体脂质纳米粒可抑制TGF-β1诱导的大鼠血管平滑肌细胞增殖,其作用机制与阻滞血管平滑肌细胞G0/G1期向S期转化及下调c-myc、c-Fos蛋白表达有关.

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abstractsObjective To observe the effect of Rhy-SLN on the proliferation of rat vascular smooth muscle cells (VSMC) induced by TGF-β1, and explore the mechanism. Methods The primary culture of rat thoracic aortic vascular smooth muscle cells was studied by tissue block culture method. The cells were divided into the control group, TGF-β1 group, TGF-β1+ the high, medium and low dosage groups of Rhy-SLN. In addition to the control group, the cells of the other groups were involved in the intervention of TGF-β1 of 20 g/L, and the high, medium and low dosage groups of Rhy-SLN cells were involved in the intervention of 25, 50, 100 mg/L of the hook teng solid lipid nanoparticles. After 24 hours of culture, MTT assay was used to detect cell proliferation inhibition rate in each group, and the cell cycle was detected by flow cytometry. The expression of c-myc and c-Fos protein in each group was detected by Western blot method. Results Compared with the TGF-β1 group, the absorbance value (0.457 ± 0.046 vs. 0.975 ± 0.049) of TGF-β1+ rhy-sln high dose group significantly decreased (P<0.01); the number of S phase cells (15.87% ± 2.47%, 15.23% ± 1.69%, 17.02% ± 2.87% vs.38.58% ± 2.68%)of TGF-β1+rhy-sln in each dose group significantly decreased(P<0.01);The c-myc(48.65 ±3.65,50.69 ± 4.16,55.29 ± 3.67 vs.68.21 ± 3.25)and c-Fos(38.78 ± 4.25,43.56 ± 3.69,46.58 ± 3.57 vs.66.54 ± 4.09) of the TGF-β1+ rhy-sln each dose group significantly decreased (P<0.01). Conclusions The Rhy-SLN can inhibit the proliferation of VSMC in rats induced by TGF-β1.Its mechanism is related to the conversion of G0/G1 phase to the S phase and the expression of the reduction of c-myc and c-fos protein.

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