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白藜芦醇抗肝星状细胞活性和抗肝纤维化的实验研究
백려호순항간성상세포활성화항간섬유화적실험연구
The effect of resveratrol on hepatic stellate cells and liver fibrogensis

万方数据

中华普通外科杂志中華普通外科雜誌 중화보통외과잡지
CHINESE JOURNAL OF GENERAL SURGERY
2013年 6期 448-451 ,共4页

俞富祥%李扬扬%朱千东%符均惠%张启瑜

유부상%리양양%주천동%부균혜%장계유

肝硬化%动物实验%白藜芦醇%肝星状细胞肝硬化%動物實驗%白藜蘆醇%肝星狀細胞
간경화%동물실험%백려호순%간성상세포
Liver cirrhosis%Animal experimentation%Resveratrol%Hepatic stellate cells

目的探讨白藜芦醇调节肝星状细胞(hepatic stellate cells,HSCs)活性及其抗肝纤维化作用.方法从大鼠肝脏分离纯化培养HSCs；DCFH-DA法检测不同浓度白藜芦醇对HSCs中活性氧的影响；通过CCK-8比色法检测白藜芦醇对HSCs增殖的影响.Western blot检测HSCs的α-肌动蛋白(α-SMA)表达.通过PCR检测白藜芦醇对HSCs活性相关基因表达的影响.给大鼠肝纤维化模型经腹输注白藜芦醇,检测肝组织病理切片,肝纤维化指标.结果从大鼠活体分离培养HSCs.白藜芦醇可抑制HSCs中活性氧的产生；明显抑制HSCs的α-SMA表达(103 ±7,90 ±7,63 ±4,53 ±3,F=62.179,P ＜0.05)与增殖(0.536±0.052,0.411±0.047,0.327±0.063,0.312±0.032,F=12.776,P＜0.05)；抑制HSCs活性相关基因(大鼠生肌调节因子、胶原蛋白Ⅲ及胶原蛋白Ⅰ)的表达(122 ±.5,96±3,68 ±3,60 ±3,F=180.600,P ＜0.05) (100 ±8,82±3,53±3,51 ±2,F =77.451,P＜0.05) (170±3,147±4,92 ±3,90 ±2,F=462.878,P＜0.05).大鼠活体实验显示白藜芦醇可降低肝羟脯氨酸含量及血清胶原蛋白Ⅲ和透明质酸水平(358.3 ±20.2,320.5±15.3,290.3±24.5,F=23.929,P ＜0.05) (32.8±3.1,28.9±1.3,25.3±1.8,F=20.050,P＜0.05)(276.3±17.8,225.3±28.3,195.4±11.2,F=18.585,P＜0.05).结论白藜芦醇能够抑制大鼠HSCs的活化增殖,对活体肝纤维化具有一定的抑制作用,这可能与白藜芦醇的抗氧化及抑制MyoD的表达作用有关.
목적 탐토백려호순조절간성상세포(hepatic stellate cells,HSCs)활성급기항간섬유화작용.방법 종대서간장분리순화배양HSCs；DCFH-DA법검측불동농도백려호순대HSCs중활성양적영향；통과CCK-8비색법검측백려호순대HSCs증식적영향.Western blot검측HSCs적α-기동단백(α-SMA)표체.통과PCR검측백려호순대HSCs활성상관기인표체적영향.급대서간섬유화모형경복수주백려호순,검측간조직병리절편,간섬유화지표.결과 종대서활체분리배양HSCs.백려호순가억제HSCs중활성양적산생；명현억제HSCs적α-SMA표체(103 ±7,90 ±7,63 ±4,53 ±3,F=62.179,P ＜0.05)여증식(0.536±0.052,0.411±0.047,0.327±0.063,0.312±0.032,F=12.776,P＜0.05)；억제HSCs활성상관기인(대서생기조절인자、효원단백Ⅲ급효원단백Ⅰ)적표체(122 ±.5,96±3,68 ±3,60 ±3,F=180.600,P ＜0.05) (100 ±8,82±3,53±3,51 ±2,F =77.451,P＜0.05) (170±3,147±4,92 ±3,90 ±2,F=462.878,P＜0.05).대서활체실험현시백려호순가강저간간포안산함량급혈청효원단백Ⅲ화투명질산수평(358.3 ±20.2,320.5±15.3,290.3±24.5,F=23.929,P ＜0.05) (32.8±3.1,28.9±1.3,25.3±1.8,F=20.050,P＜0.05)(276.3±17.8,225.3±28.3,195.4±11.2,F=18.585,P＜0.05).결론 백려호순능구억제대서HSCs적활화증식,대활체간섬유화구유일정적억제작용,저가능여백려호순적항양화급억제MyoD적표체작용유관.
Objective To study the protective effects of resveratrol against hepatic stellate cells (HSCs) and liver fibrogensis.Methods HSCs were isolated from liver of SD rats.The reactive oxygen output in HSCs under resveratrol in different concentrations was tested by DCFH-DA kit.The proliferation of HSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of HSCs was evaluated by Western blotting.The activity-related genes were measured by PCR.The models of liver fibrogenes were established.Resveratrol in different concentrations was administrated intraperitoneally.Liver was studied by pathology and SMA staining.Hydroxyproline content of liver and levels of collagen Ⅲ and hyaluronic acid in serum were tested.Results HSCs were isolated from liver and cultured successfully.Resveratrol inhibited the generation of the reactive oxygen.Proliferation and activation of HSCs was inhibited by resveratrol (0.536 ±0.052,0.411 ±0.047,0.327 ±0.063,0.312 ±0.032,F =12.776,P ＜0.05) (103 ±7,90 ±7,63 ± 4,53 ± 3,F =62.179,P ＜ 0.05).Resveratrol inhibited the expression of genes (myogenic determination gene MyoD,collagen 11 and collagen Ⅰ) in HSCs(122 ± 5,96 ± 3,68 ± 3,60 ± 3,F =180.600,P＜0.05) (100±8,82 ±3,53 ±3,51 ±2,F=77.451,P ＜0.05) (170 ±3,147 ±4,92 ±3,90 ±2,F =462.878,P ＜ 0.05).Resveratrol downregulated the level of hydroxyproline,collagen Ⅲ and hyaluronic acid (358.3 ± 20.2,320.5 ± 15.3,290.3 ± 24.5,F =23.929,P ＜ 0.05) (32.8 ± 3.1,28.9 ±1.3,25.3±1.8,F=20.050,P＜0.05)(276.3 ±17.8,225.3 ±28.3,195.4 ±11.2,F=18.585,P＜0.05).Conclusions Resveratrol can inhibit the proliferation and activation of HSCs and downregulate the fibrogensis level of the liver of rats.