茶多酚對紫外線誘導(dǎo)大鼠晶狀體上皮細胞中Caveolin-1、MMP-2表達的影響
本文選題:茶多酚 切入點:小窩蛋白-1 出處:《遼寧醫(yī)學(xué)院》2012年碩士論文 論文類型:學(xué)位論文
【摘要】:目的 研究小窩蛋白-1(caveolin-1, CAV-1)和基質(zhì)金屬蛋白酶-2(matrixmetalloproteinase-2,MMP-2)在氧化損傷性白內(nèi)障的晶狀體上皮細胞(lensepithelial cells, LECs)中的表達,并探討茶多酚(tea polyphenols,,TP)對晶狀體上皮細胞中CAV-1和MMP-2表達的影響,為探索TP可能成為防治白內(nèi)障的新藥物提供實驗基礎(chǔ)和理論依據(jù)。 方法 采用離體晶狀體培養(yǎng)技術(shù),通過紫外線(ultra-violet,UV)照射氧化損傷法建立實驗性白內(nèi)障晶狀體模型,設(shè)置空白對照組、紫外線照射組和實驗組(即UV+TP組),實驗組按茶多酚的濃度分為4個亞組,即0.02mg/L、0.2mg/L、2mg/L、20mg/L,分別于6h、12h、24h、48h取樣,進行形態(tài)學(xué)觀察,運用免疫組織化學(xué)法染色計數(shù)晶狀體上皮細胞中CAV-1、MMP-2陽性細胞,并運用Western-blot免疫印跡分析晶狀體上皮細胞中CAV-1、MMP-2表達量的變化。 結(jié)果 空白對照組大鼠離體晶狀體在培養(yǎng)的48h內(nèi)無顯著變化,晶狀體均保持透明,紫外線照射組大鼠離體晶狀體在培養(yǎng)的48h內(nèi),隨紫外線作用時間的延長,晶狀體的混濁程度逐漸加重。不同濃度TP實驗組晶狀體混濁進展較UV組緩慢,在不同時間段的混濁程度均明顯低于UV組。不同時段各組晶狀體上皮細胞Caveolin-1、MMP-2陽性表達率和表達量的比較,各實驗組與紫外線對照組相比,差異有均有統(tǒng)計學(xué)意義(P0.05),不同時段實驗組晶狀體上皮細胞CAV-1陽性表達明顯高于UV組,MMP-2陽性表達明顯低于UV組,0.02mg/L TP實驗組CAV-1與空白對照組相比,差異無統(tǒng)計學(xué)意義(P0.05);其余實驗組與空白對照組相比,差異均有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論 CAV-1存在于正常晶狀體上皮細胞中,在白內(nèi)障晶狀體上皮細胞中,隨著紫外線照射時間的延長,CAV-1表達逐漸減少; MMP-2在正常晶狀體上皮細胞中無表達,在白內(nèi)障晶狀體上皮細胞中,隨著紫外線照射時間的延長,MMP-2的表達逐漸增加。TP可以抑制紫外線誘導(dǎo)的晶狀體上皮細胞CAV-1表達的減少和MMP-2的增加,以濃度為0.02mg/L TP的作用效果最佳,提示TP有可能成為防治白內(nèi)障的有效藥物。
[Abstract]:Purpose. To investigate the expression of fossa protein 1 (CAV-1) and matrix metalloproteinase-2 (MMP-2) in lens epithelial cells (LECs) of oxidative cataract, and to investigate the effect of tea polyphenols tea polyphenols (TP) on the expression of CAV-1 and MMP-2 in lens epithelial cells. In order to explore TP may be a new drug for cataract prevention and treatment of experimental basis and theoretical basis. Method. The experimental cataract lens model was established by ultraviolet ultra-violetine (UV) irradiation and oxidative injury in vitro, and a blank control group was set up. Ultraviolet irradiation group and experimental group (UV TP group) were divided into four subgroups according to the concentration of tea polyphenols. The experimental group was divided into 4 subgroups according to the concentration of tea polyphenols, namely 0.02mg / L 0.2mg / L 0.2mg / L ~ (2) mg / L ~ (2) mg / L ~ (2) mg / L ~ (20) mg 路L ~ (-1) respectively at 6h ~ 12h ~ (24) h ~ (24 h) for morphological observation, and immunohistochemical staining was used to count the CAV-1MMP ~ (2) positive cells in lens epithelial cells. The expression of CAV-1 and MMP-2 in lens epithelial cells was analyzed by Western-blot Western blot. Results. In the blank control group, there was no significant change in the isolated lens within 48 hours of culture, and the lens remained transparent, while in the ultraviolet radiation group, the lens of the isolated lens cultured within 48 hours was prolonged with the time of ultraviolet irradiation. The degree of lens opacity was gradually aggravated. The progress of lens opacity in TP groups with different concentrations was slower than that in UV group. The positive rate and quantity of Caveolin-1 MMP-2 in lens epithelial cells in different periods of time were significantly lower than those in UV group, and the positive rate and quantity of MMP-2 in each experimental group were higher than those in ultraviolet control group. The positive expression of CAV-1 in lens epithelial cells in the experimental group was significantly higher than that in the UV group (P 0.05). The positive expression of MMP-2 in the experimental group was significantly lower than that in the UV group (0.02 mg / L TP) compared with the blank control group. There was no significant difference between the two groups (P 0.05), but there was significant difference between the other experimental groups and the blank control group (P 0.05). Conclusion. The expression of CAV-1 in normal lens epithelial cells decreased gradually with the prolongation of ultraviolet irradiation time, but MMP-2 was not expressed in normal lens epithelial cells, but in cataract lens epithelial cells. With the prolongation of ultraviolet irradiation time, the expression of MMP-2 increased gradually. TP could inhibit the decrease of CAV-1 expression and the increase of MMP-2 in lens epithelial cells induced by ultraviolet radiation. The best effect was 0.02mg / L TP. The results suggest that TP may be an effective drug for the prevention and treatment of cataract.
【學(xué)位授予單位】:遼寧醫(yī)學(xué)院
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2012
【分類號】:R776.1
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