中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2010年
2期
169-172
,共4页
宋虎平%惠延年%雷春灵%王建洲%毕春潮%杨新光
宋虎平%惠延年%雷春靈%王建洲%畢春潮%楊新光
송호평%혜연년%뢰춘령%왕건주%필춘조%양신광
糖尿病视网膜病变/病理生理学%缺氧诱导因子1%亚基%细胞黏附分子/生理学%临床实验室技术/方法
糖尿病視網膜病變/病理生理學%缺氧誘導因子1%亞基%細胞黏附分子/生理學%臨床實驗室技術/方法
당뇨병시망막병변/병리생이학%결양유도인자1%아기%세포점부분자/생이학%림상실험실기술/방법
Diabetic retinopathy/Phisiopathology%Hypoxia-inducible factor 1,alpha subunit%Cell adhesion molecules/physiology%Clinical laboratory techniques/methods
目的 观察在早期糖尿病视网膜病变(DR)状态下,缺氧诱导因子1α(HIF-1α)对细胞表面黏附分子CD18表达以及白细胞和血管内皮细胞黏附的影响.方法 收集早期DR患者及年龄匹配的健康人外周血血清,用于体外培养人粒细胞系白血病细胞(HL60)和恒河猴视网膜血管内皮细胞系细胞(RF/6A).分为糖尿病血清培养组(B组)、糖尿病+HIF-1α反义寡核苷酸组(ASODN)(C组)、糖尿病+HIF-1α正义寡核苷酸组(SODN)(D组),健康人血清培养细胞为正常对照组(A组).以流式细胞仪和Real-time PCR分别检测HL60细胞表面CD18蛋白阳性细胞比例和CD18 mRNA的表达水平,以虎红染色法观察HL60细胞和RF/6A细胞的黏附率.结果 A、B、C、D组CD18阳性细胞比例分别为17.06±6.01、42.23±2.60、25.33±3.05、32.40±10.57,各组之间差异有统计学意义(F=36.47,P<0.001).和A组比较,B、C、D组CD18 mRNA水平分别是A组的21.05±2.07、2.23±0.96、25.07±2.27倍,各组之间差异有统计学意义(F=180.34,P<0.001).A、B、C、D组细胞黏附率分别为0.06±0.002、0.09±0.10、0.05±0.007、0.07±0.01,各组之间差异有统计学意义(F=13.06,P=0.002) 结论在体外培养条件下,糖尿病血清可以促进HL60细胞表达CD18蛋白和CD18mRNA,促进HL60细胞和血管内皮细胞的黏附,HIF-1α表达被抑制后,这种促进作用减弱.HIF-1α对糖尿病状态下细胞表面黏附分子CD18表达及白细胞和血管内皮细胞之间的黏附具有调节作用.
目的 觀察在早期糖尿病視網膜病變(DR)狀態下,缺氧誘導因子1α(HIF-1α)對細胞錶麵黏附分子CD18錶達以及白細胞和血管內皮細胞黏附的影響.方法 收集早期DR患者及年齡匹配的健康人外週血血清,用于體外培養人粒細胞繫白血病細胞(HL60)和恆河猴視網膜血管內皮細胞繫細胞(RF/6A).分為糖尿病血清培養組(B組)、糖尿病+HIF-1α反義寡覈苷痠組(ASODN)(C組)、糖尿病+HIF-1α正義寡覈苷痠組(SODN)(D組),健康人血清培養細胞為正常對照組(A組).以流式細胞儀和Real-time PCR分彆檢測HL60細胞錶麵CD18蛋白暘性細胞比例和CD18 mRNA的錶達水平,以虎紅染色法觀察HL60細胞和RF/6A細胞的黏附率.結果 A、B、C、D組CD18暘性細胞比例分彆為17.06±6.01、42.23±2.60、25.33±3.05、32.40±10.57,各組之間差異有統計學意義(F=36.47,P<0.001).和A組比較,B、C、D組CD18 mRNA水平分彆是A組的21.05±2.07、2.23±0.96、25.07±2.27倍,各組之間差異有統計學意義(F=180.34,P<0.001).A、B、C、D組細胞黏附率分彆為0.06±0.002、0.09±0.10、0.05±0.007、0.07±0.01,各組之間差異有統計學意義(F=13.06,P=0.002) 結論在體外培養條件下,糖尿病血清可以促進HL60細胞錶達CD18蛋白和CD18mRNA,促進HL60細胞和血管內皮細胞的黏附,HIF-1α錶達被抑製後,這種促進作用減弱.HIF-1α對糖尿病狀態下細胞錶麵黏附分子CD18錶達及白細胞和血管內皮細胞之間的黏附具有調節作用.
목적 관찰재조기당뇨병시망막병변(DR)상태하,결양유도인자1α(HIF-1α)대세포표면점부분자CD18표체이급백세포화혈관내피세포점부적영향.방법 수집조기DR환자급년령필배적건강인외주혈혈청,용우체외배양인립세포계백혈병세포(HL60)화항하후시망막혈관내피세포계세포(RF/6A).분위당뇨병혈청배양조(B조)、당뇨병+HIF-1α반의과핵감산조(ASODN)(C조)、당뇨병+HIF-1α정의과핵감산조(SODN)(D조),건강인혈청배양세포위정상대조조(A조).이류식세포의화Real-time PCR분별검측HL60세포표면CD18단백양성세포비례화CD18 mRNA적표체수평,이호홍염색법관찰HL60세포화RF/6A세포적점부솔.결과 A、B、C、D조CD18양성세포비례분별위17.06±6.01、42.23±2.60、25.33±3.05、32.40±10.57,각조지간차이유통계학의의(F=36.47,P<0.001).화A조비교,B、C、D조CD18 mRNA수평분별시A조적21.05±2.07、2.23±0.96、25.07±2.27배,각조지간차이유통계학의의(F=180.34,P<0.001).A、B、C、D조세포점부솔분별위0.06±0.002、0.09±0.10、0.05±0.007、0.07±0.01,각조지간차이유통계학의의(F=13.06,P=0.002) 결론재체외배양조건하,당뇨병혈청가이촉진HL60세포표체CD18단백화CD18mRNA,촉진HL60세포화혈관내피세포적점부,HIF-1α표체피억제후,저충촉진작용감약.HIF-1α대당뇨병상태하세포표면점부분자CD18표체급백세포화혈관내피세포지간적점부구유조절작용.
Objective To observe the effect of hypoxia inducible factor-1α(HIF-1)to the expression of cell surface adhesion molecules CD18 and the adhesion ability of leukoeytes and vascular endothelial cells under early stage of diabetic retinopathy condition. Methods The human promyeloeytic leukemia cell line HL60 and the rhesus choroid-retina vascular endothelial cell line RF/6A were cultured in RPMI 1640 medium-10% human serum, which was collected from the subjects of early stage of diabetic retinopathy and age-matched healthy control. The cells were cultured in 4 groups as control group (group A), diabetic group (group B), HIF-1 anti-sense oligonucleotides (ASODN) group (group C) and HIF-1 sense oligonueleotides (SODN) group (group D). The percentages of CD18 positive cell in the HL60 cell were measured by flow cytometry and mRNA in the HL60 cell by real-time reverse transcription-polymerase chain reaction(RT-PCR). Results The percentage of CD18 positive cell in the group A, B, C and D was 17.06±6.01, 42. 23±2.60, 25.33±3.05 and 32.40±10.57, respectively, the differences among them were significant (F=36.47,P<0. 001). Compared to the group A, the expression of CD18 mRNA in the group B, C and D was increased about 21. 05±2. 07,2. 23±0.96 and 25.07±2.27 times, respectively, the differences among them were significant (F=180.34, P<0. 001). The adherent rates of HL60 to RF/6A in group A, B, C and D was 0. 06±0. 00, 0. 09±0. 10, 0. 05 ±0. 00 and 0.07 ± 0. 01, respectively, the differences among them were significant(F=13.06,P=0. 002). Conclusion In vitro, HIF-1 could regulate the expression of CD18 by HL60, and the adhesion of HL60 to RF/6A when the cells were exposed to diabetic serum. The effects of human serum weaken with the inhibition of HIF-1 expression. HIF-1 play regulatory role in the expression of CD18 and adhesion of leukoeytes and vascular endothelial cells under early stage of diabetic retinopathy condition.
