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  本文選題：蘋果酸舒尼替尼　切入點：恒河猴脈絡(luò)膜視網(wǎng)膜血管內(nèi)皮細(xì)胞　出處：《廣西醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 目的探討蘋果酸舒尼替尼(Sunitinib malate, SU11248)對體外培養(yǎng)的眼底微血管內(nèi)皮細(xì)胞(恒河猴脈絡(luò)膜視網(wǎng)膜血管內(nèi)皮細(xì)胞,RF／6A)的增殖和遷移及KDRmRNA表達(dá)的影響。 方法體外培養(yǎng)RF／6A,分為S0、S1、S2、S3、S4、S5、S6組(各組培養(yǎng)基中Sunitinib的濃度分別為0 mg·L-1,0.00125 mg·L-1、0.0025 mg·L-1、0.005 mg-L-1、0.01 mg·L-1、0.02 mg-L-1和0.04 mg·L-1)。分別于培養(yǎng)24 h和48 h后采用磺酰羅丹明染色法(SRB)法觀察0 mg·L-1、0.00125mg·L-1、0.0025 mg·L-1、0.005 mg·L-1、0.01 mg·L-1、0.02 mg·L-1和0.04 mg·L-1濃度的Sunitinib對RF／6A增殖的影響；并根據(jù)吸光度值(OD570)計算Sunitinib對RF／6A增殖抑制的抑制率；采用細(xì)胞劃痕修復(fù)實驗觀察0 mg·L-1、0.0025 mg·L-1、0.005 mg·L-1、0.01 mg·L-10.02 mg·L-1濃度的Sunitinib對RF／6A遷移的影響；并用倒置相差顯微鏡觀察RF／6A遷移的情況；同時利用逆轉(zhuǎn)錄—聚合酶鏈?zhǔn)椒磻?yīng)(reverse transcription-polymerase chain reaction, RT-PCR)檢測0 mg·L-1、0.0025mg·L-1、0.005 mg·L-1、0.01 mg·L-1、0.02 mg·L-1濃度的Sunitinib處理前后RF/6A KDR (VEGFR-2)mRNA表達(dá)水平的變化。各組RF／6A吸光度A值,RF／6A增殖抑制的抑制率及KDRmRNA相對含量均以均數(shù)±標(biāo)準(zhǔn)差(x±s)表示,數(shù)據(jù)處理采用SPSS13.0統(tǒng)計軟件進(jìn)行單因素方差分析(one-way ANOVA),并采用LSD法進(jìn)行兩兩比較,P0.05具有統(tǒng)計學(xué)意義。 結(jié)果1 SRB實驗結(jié)果如下： Sunitinib對RF／6A的增殖有抑制作用且呈時間—劑量依賴性,0.00125 mg·L-1、0.0025 mg·L-1、0.005 mg·L-1、0.01 mg·L-1、0.02 mg·L-1和0.04 mg·L-1濃度的Sunitinib作用于RF／6A 24 h,抑制率分別為(12.009±0.038)%、(21.440±0.007)%、(35.434±0.015)%、(43.125±0.002)%、(53.700±0.001)%、(60.971±0.003)%,各組兩兩比較差異有統(tǒng)計學(xué)意義(P0.01)；Sunitinib作用于RF／6A48 h,其抑制率分別為(36.872±0.006)%、(40.673±0.013)%、(47.313±0.004)%、(55.910±0.003)%、(63.120±0.003)%、(69.975±0.014)%,各組兩兩比較差異均有統(tǒng)計學(xué)意義(P0.01)。 2細(xì)胞遷移劃痕修復(fù)實驗結(jié)果如下： 培養(yǎng)基中加入0 mg·L-1、0.0025 mg·L-1、0.005 mg·L-1、0.01mg·L-1、0.02 mg·L-1濃度的Sunitinib 24 h后RF／6A的遷移距離分別為(203.3±2.2)μm、(145.4±4.4)μm、(123.9±2.6)μm、(96.1±3.1)μm、(46.6±2.9)μm。而48 h后則為(313.1±4.1)μm、(213.9±2.8)μm、(193.9±4.2)μm、(134.5±3.2)μm、(109.9±5.7)μm。在同一時段,各組兩兩比較差異均有統(tǒng)計學(xué)意義(P0.01)。 3 RT-PCR結(jié)果如下： 0 mg·L-1、0.0025 mg·L-1、0.005 mg·L-1、0.01 mg·L-1、0.02mg·L-1濃度的Sunitinib作用24 h后,RF/6A KDRmRNA的相對表達(dá)量分別為0.583±0.004、0.570±0.008、0.553±0.007、0.531±0.003、0.513±0.005。而48 h后則分別為0.628±0.005、0.610±0.002、0.588±0.002、0.564±0.005、0.525±0.004。各組兩兩比較差異均有統(tǒng)計學(xué)意義(P0.01)。 結(jié)論1Sunitinib能夠抑制體外培養(yǎng)的恒河猴脈絡(luò)膜視網(wǎng)膜血管內(nèi)皮細(xì)胞的增殖和遷移；Sunitinib能夠下調(diào)恒河猴脈絡(luò)膜視網(wǎng)膜血管內(nèi)皮細(xì)胞KDRmRNA的表達(dá)； 2 Sunitinib抑制視網(wǎng)膜血管內(nèi)皮細(xì)胞的增殖和遷移的機(jī)制可能與其下調(diào)VEGFR的表達(dá)有關(guān)；
[Abstract]:Objective to investigate the sunitinib malate (Sunitinib, malate, SU11248) on cultured retinal microvascular endothelial cells (endothelial cells, Ganges RIver monkey choroidal retinal RF / 6A) expression of proliferation and migration and KDRmRNA.
RF / 6A culture method in vitro, divided into S0, S1, S2, S3, S4, S5, S6 group (the concentration of Sunitinib in the culture medium of each group were 0 mg - L-1,0.00125 Mg - L-1,0.0025 Mg - L-1,0.005 mg-L-1,0.01 Mg - L-1,0.02 mg-L-1 and 0.04 mg L-1). The staining was cultured for 24 h and 48 h after using sulfonyl Luo Danming (SRB) Mg - L-1,0.00125mg - L-1,0.0025 0 mg - L-1,0.005 Mg - L-1,0.01 Mg - L-1,0.02 Mg - L-1 and 0.04 mg L-1 concentration of Sunitinib on RF / 6A proliferation observation; and according to the absorbance value (OD570) of RF / Sunitinib computing to inhibit 6A proliferation rate; by cell scratch repair experimental observation of 0 mg - L-1,0.0025 Mg - L-1,0.005 Mg - L-1,0.01 Mg - L-10.02 Mg - L-1 concentration effect of Sunitinib on RF / 6A migration; and observed by phase contrast RF / 6A migration of the microscope; at the same time by using reverse transcription polymerase chain reaction (reverse, tra Nscription-polymerase chain reaction, RT-PCR Sunitinib) before and after treatment was detected in 0 mg - L-1,0.0025mg - L-1,0.005 Mg - L-1,0.01 Mg - L-1,0.02 Mg - L-1 concentration of RF/6A KDR (VEGFR-2) expression of mRNA. Each RF / 6A value A, the relative content of KDRmRNA RF / 6A and the inhibition rate of proliferation are mean standard the difference (x + s), data processing using SPSS13.0 statistical software for single factor analysis of variance (one-way ANOVA), and LSD method was used to compare the 22, P0.05 was statistically significant.
Results 1 SRB experimental results are as follows:
Sunitinib on RF / 6A inhibited the proliferation and showed time and dose dependent, 0.00125 Mg - L-1,0.0025 Mg - L-1,0.005 Mg - L-1,0.01 Mg - L-1,0.02 Mg - L-1 and 0.04 mg L-1 concentration of Sunitinib in RF / 6A 24 h, the inhibition rate of (12.009 + 0.038)%, (21.440. 0.007)% and (35.434 + 0.015)% and (43.125 + 0.002)% and (53.700 + 0.001)% and (60.971 + 0.003)%, 22 groups was statistically significant difference (P0.01); the effect of Sunitinib on RF / 6A48 h, and the inhibition rates were (36.872 + 0.006)%, (40.673 + 0.013)% and (47.313 + 0.004)% and (55.910 + 0.003)% and (63.120 + 0.003)% and (69.975 + 0.014)%, 22 groups were statistically significant (P0.01).
2 cell migration scratch repair experimental results are as follows:
Adding 0 mg - L-1,0.0025 culture mg L-1,0.005 mg L-1,0.01mg L-1,0.02 Mg - L-1 concentration in medium Sunitinib 24 h after the migration distance of RF / 6A respectively (203.3 + 2.2) m, (145.4 + 4.4) m, (123.9 + 2.6) m, (96.1 + 3.1) m, (46.6 + 2.9) M. and 48 h for (313.1 + 4.1) m, (213.9 + 2.8) m, (193.9 + 4.2) m, (134.5 + 3.2) m, (109.9 + 5.7) M. at the same time, the 22 groups had statistical difference meaning (P0.01).
3 RT-PCR results are as follows:
Sunitinib 0 mg - L-1,0.0025 Mg - L-1,0.005 Mg - L-1,0.01 Mg - L-1,0.02mg - L-1 concentration of 24 h, the relative expression of RF/6A KDRmRNA were 0.583 + 0.004,0.570 + 0.008,0.553 + 0.007,0.531 + 0.003,0.513 + 0.005. and 48 h respectively for 0.628 + 0.005,0.610 + 0.002,0.588 + 0.002,0.564 + 0.005,0.525 + 0.004. were 22 difference there was statistically significant (P0.01).
The proliferation and migration of Ganges RIver monkey choroid retinal endothelial cells in vitro. Conclusion 1Sunitinib can inhibit the expression of Sunitinib can down regulate endothelial cells; KDRmRNA Ganges RIver monkey choroidal retinal vessels;
The mechanism of 2 Sunitinib inhibit the proliferation and migration of retinal vascular endothelial cells may be associated with down-regulation of VEGFR;

【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R774

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本文編號：1692114