發(fā)布時(shí)間：2018-07-10 17:08

  本文選題：雌二醇 + 晶狀體上皮細(xì)胞　； 參考：《瀘州醫(yī)學(xué)院》2010年碩士論文

【摘要】：目的：觀察雌激素中活性最強(qiáng)的17β-雌二醇(17β-estradiol, 17β-E2)在不同濃度時(shí)對過氧化氫(hydrogen dioxide, H2O2)誘導(dǎo)的雌性大鼠晶狀體上皮細(xì)胞凋亡的影響及其對凋亡相關(guān)蛋白Bcl-2和Bax表達(dá)的調(diào)控,探討不同濃度雌激素對晶狀體上皮細(xì)胞凋亡的作用及其機(jī)制,為合理應(yīng)用雌激素防治白內(nèi)障提供實(shí)驗(yàn)依據(jù)。方法：摘取健康成年雌性SD大鼠的透明晶狀體48枚,并置于Medium 199培養(yǎng)液中培養(yǎng)。按培養(yǎng)液中添加的成分差異隨機(jī)分為6組：17β-E2+H2O2組1、17β-E2+H2O2組2、17β-E2+H2O2組3、17β-E2+H2O2組4、H2O2組以及空白對照組。予以終濃度為300μmol/L的過氧化氫復(fù)制SD大鼠晶狀體上皮細(xì)胞凋亡模型,分別加入不同終濃度的17β-雌二醇干預(yù)17β-E2+H2O2組1至4(干預(yù)濃度分別為：1×10-8mol/L, 1×10-7mol/L, 1×10-6mol/L, 1×10-5mol/L。于CO2細(xì)胞恒溫培養(yǎng)箱中共同孵育24小時(shí)后觀察并比較各組晶狀體混濁情況；每組取2枚晶狀體行石蠟包埋、切片、蘇木素-伊紅染色(hematoxylin and eosin stain, HE stain);撕取各組剩余晶狀體(6枚/組)前囊及赤道部囊膜鋪片,采用脫氧核糖核酸末端轉(zhuǎn)移酶缺口標(biāo)記原位細(xì)胞凋亡檢測法(terminal deoxynucleotidyl transferasa(TdT)-medinted dUTP nick-end labeling, TUNEL法)檢測晶狀體上皮細(xì)胞凋亡,鏈霉菌抗生物素蛋白-過氧化物酶連接法(Streptavidin Peroxidase Conjugated Method, SP法)檢測凋亡相關(guān)蛋白Bcl-2和Bax的表達(dá)并進(jìn)行比較。統(tǒng)計(jì)學(xué)分析：SPSS13.0單因素方差分析,并選用LSD法及SNK法進(jìn)行兩兩比較。結(jié)果：經(jīng)24小時(shí)孵育,空白對照組晶狀體完全透明,H2O2組及17β-E2+H2O2各組晶狀體混濁,但17β-E2+H2O2各組晶狀體混濁程度均低于H2O2組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)；TUNEL法檢測細(xì)胞凋亡結(jié)果顯示：終濃度為300μmol/L的過氧化氫能成功誘導(dǎo)晶狀體上皮細(xì)胞凋亡,17β-E2+H2O2各組晶狀體上皮細(xì)胞凋亡率均顯著低于H2O2組,差異具有統(tǒng)計(jì)學(xué)意義(P0.05),且與17β-雌二醇的濃度呈負(fù)相關(guān)(均數(shù)圖Means Plots)。免疫組化SP法檢測Bcl-2及Bax陽性表達(dá)率結(jié)果顯示：在空白對照組晶狀體上皮細(xì)胞中,Bcl-2和Bax均有表達(dá),且Bcl-2的表達(dá)遠(yuǎn)較Bax表達(dá)強(qiáng),Bcl-2/Bax比值處于較高水平；過氧化氫可明顯降低Bcl-2表達(dá),提高Bax表達(dá)(P0.05),Bcl-2/Bax比值下降；與H2O2組比較,17β-雌二醇可明顯提高Bcl-2表達(dá),降低Bax表達(dá)(P0.05),Bcl-2/Bax比值升高,且在一定范圍內(nèi)與17β-雌二醇濃度呈正相關(guān)(均數(shù)圖Means Plots)。結(jié)論：17β-雌二醇對過氧化氫誘導(dǎo)的雌性大鼠晶狀體上皮細(xì)胞凋亡有抑制作用,此抑制作用在一定范圍內(nèi)與17β-雌二醇的濃度呈正相關(guān)。對凋亡相關(guān)蛋白Bcl-2和Bax表達(dá)的調(diào)控可能是17β-雌二醇抑制晶狀體上皮細(xì)胞凋亡的分子機(jī)制之一。
[Abstract]:Aim: to investigate the effects of 17 尾 -estradiol (17 尾 -E2), the most active estrogen, on the apoptosis of female rat lens epithelial cells induced by hydrogen peroxide (hydrogen dioxide, H _ 2O _ 2) at different concentrations and the regulation of Bcl-2 and Bax expression. To investigate the effect of estrogen at different concentrations on lens epithelial cell apoptosis and its mechanism, and to provide experimental evidence for the rational use of estrogen in the prevention and treatment of cataract. Methods: 48 clear lenses of healthy adult female SD rats were obtained and cultured in medium 199 medium. According to the difference of the components added in the culture medium, they were randomly divided into 6 groups: 1 / 17 尾 -E2 H _ 2O _ 2 group (1 / 17 尾 -E _ 2) H _ 2O _ 2 group ~ (2) ~ (2) ~ (17) 尾 -E _ (2) H _ 2O _ (2) _ (3) H _ 2O _ 2 group (n = 4) and blank control group (n = 6). SD rat lens epithelial cell apoptosis model was induced by hydrogen peroxide at the final concentration of 300 渭 mol / L, and 17 尾 -estradiol at different final concentrations was added to treat 17 尾 -E2 H _ 2O _ 2 group 1 to 4 (1 脳 10 ~ (-8) mol / L, 1 脳 10 ~ (-7) mol / L, 1 脳 10 ~ (-6) mol 路L ~ (-1), 1 脳 10 ~ (-5) mol / L) respectively. After incubating in a CO _ 2 cell incubator for 24 hours, the lens opacity in each group was observed and compared. (hematoxylin and eosin stain, HE stain); stained with hematoxylin and eosin were used to tear the anterior capsule and equatorial capsule of the remaining lens (6 / group). Apoptosis of lens epithelial cells was detected by deoxyribonucleic acid terminal transferase Nick labeling (terminal deoxynucleotidyl transferasa (TDT) -meditated dUTP nick-end labeling (Tunel method). Streptavidin Peroxidase conjugated method (SP) was used to detect the expression of apoptosis-related proteins Bcl-2 and Bax. The single factor ANOVA was analyzed by: SPSS 13.0, and the LSD method and SNK method were used to carry out the comparison. Results: after 24 hours incubation, the lens opacity in the blank control group was completely transparent and transparent, but the degree of lens opacity in 17 尾 -E2 H 2O 2 group was lower than that in H 2O 2 group (P0.05). The results of Tunel assay showed that hydrogen peroxide with final concentration of 300 渭 mol / L could successfully induce apoptosis of lens epithelial cells. The apoptosis rate of lens epithelial cells in H _ 2O _ 2 group was significantly lower than that in H _ 2O _ 2 group. The difference was statistically significant (P0.05) and negatively correlated with the concentration of 17 尾 -estradiol (mean means lots). The expression of Bcl-2 and Bax in lens epithelial cells of blank control group was detected by immunohistochemical SP method. The expression of Bcl-2 and Bax was much higher than that of Bax, and hydrogen peroxide could significantly decrease the expression of Bcl-2. Compared with H2O2 group, 17 尾 -estradiol significantly increased Bcl-2 expression, decreased Bax expression (P0.05) and increased Bcl-2 / Bax ratio, and was positively correlated with 17 尾 -estradiol concentration in a certain range (mean figure means lots). Conclusion the effect of 1: 17 尾 -estradiol on the apoptosis of female rat lens epithelial cells induced by hydrogen peroxide is positively correlated with the concentration of 17 尾 -estradiol. The regulation of apoptosis-related proteins Bcl-2 and Bax may be one of the molecular mechanisms of 17 尾 -estradiol inhibiting the apoptosis of lens epithelial cells.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R776.1

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本文編號：2114043