中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2014年
2期
156-160
,共5页
宋虎平%朱琦%吴琼%艾华%雷哓琴%朱昭亮%杨阳
宋虎平%硃琦%吳瓊%艾華%雷嘵琴%硃昭亮%楊暘
송호평%주기%오경%애화%뢰효금%주소량%양양
糖尿病视网膜病变/病理生理学%缺氧诱导因子1,α亚基%RNA,小分子干扰
糖尿病視網膜病變/病理生理學%缺氧誘導因子1,α亞基%RNA,小分子榦擾
당뇨병시망막병변/병리생이학%결양유도인자1,α아기%RNA,소분자간우
Diabetic retinopathy/pathophsiology%Hypoxia-inducible factor 1,alpha subunit%RNA,small interfering
目的 观察早期糖尿病视网膜病变(DR)状态下,以pSUPER为载体的缺氧诱导因子1α(HIF1α)特异性小干扰(siHIF1α)RNA对髓样细胞表面黏附分子CD18及神经损伤诱导蛋白-1(Ninj-1)表达的影响.方法 实验分为健康人血清培养组(A组)、糖尿病血清培养组(B组)、糖尿病血清培养联合pSUPERH1-siHIF1α转染组(C组)及糖尿病血清培养联合pSUPER空载体组(D组)进行.A组采用健康志愿者血清培养人髓样细胞系白血病细胞系(K562)细胞;B、C、D组均采用DR患者血清培养K562细胞.C、D组在加入DR患者血清前24 h分别转染pSUPERH1-siHIF1α及pSUPER空载体.采用流式细胞仪检测各组K562细胞表面的CD18、Ninj-1表达.行细胞黏附实验,检测各组K562细胞与恒河猴视网膜脉络膜血管内皮细胞系(RF/6A)细胞黏附率.结果 A、B、C、D组K562细胞表面CD18表达比较,4组间差异有统计学意义(F=14.33,P=0.01).组间CD 18表达两两比较,B组明显高于A组(P=0.001);C组明显低于B组(P=0.001)和D组(P=0.02);C组与A组间无明显差异(95％可信区间=-14.89～2.13,P=0.12).A、B、C、D组K562细胞表面Ninj-1表达比较,4组间差异有统计学意义(F=39.38,P=0.001).组间Ninj-1表达两两比较,B组明显高于A组(P=0.00);C组与B组间无明显差异(P=0.06);C组与D组间也无明显差异(P=0.49).A、B、C、D组K562细胞与RF/6A细胞黏附率比较,4组间差异有统计学意义(F=20.62,P=0.00).组间K562细胞与RF/6A细胞黏附率两两比较,B组明显高于A组(P=0.00);C组较B组明显下降(P=0.01),较A组明显提高(P=0.002);B组与D组间无明显差异(P=0.68).结论 早期DR状态下,以pSUPER为载体的siHIF1α RNA可降低K562细胞表面CD18的表达,但对Ninj-1表达无明显影响.
目的 觀察早期糖尿病視網膜病變(DR)狀態下,以pSUPER為載體的缺氧誘導因子1α(HIF1α)特異性小榦擾(siHIF1α)RNA對髓樣細胞錶麵黏附分子CD18及神經損傷誘導蛋白-1(Ninj-1)錶達的影響.方法 實驗分為健康人血清培養組(A組)、糖尿病血清培養組(B組)、糖尿病血清培養聯閤pSUPERH1-siHIF1α轉染組(C組)及糖尿病血清培養聯閤pSUPER空載體組(D組)進行.A組採用健康誌願者血清培養人髓樣細胞繫白血病細胞繫(K562)細胞;B、C、D組均採用DR患者血清培養K562細胞.C、D組在加入DR患者血清前24 h分彆轉染pSUPERH1-siHIF1α及pSUPER空載體.採用流式細胞儀檢測各組K562細胞錶麵的CD18、Ninj-1錶達.行細胞黏附實驗,檢測各組K562細胞與恆河猴視網膜脈絡膜血管內皮細胞繫(RF/6A)細胞黏附率.結果 A、B、C、D組K562細胞錶麵CD18錶達比較,4組間差異有統計學意義(F=14.33,P=0.01).組間CD 18錶達兩兩比較,B組明顯高于A組(P=0.001);C組明顯低于B組(P=0.001)和D組(P=0.02);C組與A組間無明顯差異(95％可信區間=-14.89～2.13,P=0.12).A、B、C、D組K562細胞錶麵Ninj-1錶達比較,4組間差異有統計學意義(F=39.38,P=0.001).組間Ninj-1錶達兩兩比較,B組明顯高于A組(P=0.00);C組與B組間無明顯差異(P=0.06);C組與D組間也無明顯差異(P=0.49).A、B、C、D組K562細胞與RF/6A細胞黏附率比較,4組間差異有統計學意義(F=20.62,P=0.00).組間K562細胞與RF/6A細胞黏附率兩兩比較,B組明顯高于A組(P=0.00);C組較B組明顯下降(P=0.01),較A組明顯提高(P=0.002);B組與D組間無明顯差異(P=0.68).結論 早期DR狀態下,以pSUPER為載體的siHIF1α RNA可降低K562細胞錶麵CD18的錶達,但對Ninj-1錶達無明顯影響.
목적 관찰조기당뇨병시망막병변(DR)상태하,이pSUPER위재체적결양유도인자1α(HIF1α)특이성소간우(siHIF1α)RNA대수양세포표면점부분자CD18급신경손상유도단백-1(Ninj-1)표체적영향.방법 실험분위건강인혈청배양조(A조)、당뇨병혈청배양조(B조)、당뇨병혈청배양연합pSUPERH1-siHIF1α전염조(C조)급당뇨병혈청배양연합pSUPER공재체조(D조)진행.A조채용건강지원자혈청배양인수양세포계백혈병세포계(K562)세포;B、C、D조균채용DR환자혈청배양K562세포.C、D조재가입DR환자혈청전24 h분별전염pSUPERH1-siHIF1α급pSUPER공재체.채용류식세포의검측각조K562세포표면적CD18、Ninj-1표체.행세포점부실험,검측각조K562세포여항하후시망막맥락막혈관내피세포계(RF/6A)세포점부솔.결과 A、B、C、D조K562세포표면CD18표체비교,4조간차이유통계학의의(F=14.33,P=0.01).조간CD 18표체량량비교,B조명현고우A조(P=0.001);C조명현저우B조(P=0.001)화D조(P=0.02);C조여A조간무명현차이(95％가신구간=-14.89～2.13,P=0.12).A、B、C、D조K562세포표면Ninj-1표체비교,4조간차이유통계학의의(F=39.38,P=0.001).조간Ninj-1표체량량비교,B조명현고우A조(P=0.00);C조여B조간무명현차이(P=0.06);C조여D조간야무명현차이(P=0.49).A、B、C、D조K562세포여RF/6A세포점부솔비교,4조간차이유통계학의의(F=20.62,P=0.00).조간K562세포여RF/6A세포점부솔량량비교,B조명현고우A조(P=0.00);C조교B조명현하강(P=0.01),교A조명현제고(P=0.002);B조여D조간무명현차이(P=0.68).결론 조기DR상태하,이pSUPER위재체적siHIF1α RNA가강저K562세포표면CD18적표체,단대Ninj-1표체무명현영향.
Objective To investigate the effects of hypoxia-inducible factor 1α (HIF-1α) small interfere RNA construct pSUPERH1-siHIF1α on the expression of CD18 and ninjurin 1 by K562 (human chronic myelogenous leukemia cell line) cells cultured with serums from patients with early stage of diabetic retinopathy.Methods K562 cells were cultured in 4 groups as control group (group A),diabetic group (group B),diabetes and pSUPERH1-siHIF1α transfect group (group C) and diabetes and pSUPER-retro transfect group (group D).The cells in group A were cultured in human serum from age-matched healthy control,and in group B,C and D,the cells were cultured in serum from the subjects of early stage of diabetic retinopathy.Twenty-four hours before the cells were cultured by the serum from the subjects of early stage of diabetic retinopathy,the HIF-1α specific siRNA expression vector pSUPERH1-siHIF1α and empty vector pSUPER-retro were transfected into the cells of group C and D,respectively.The percentages of CD18 and ninjurin-1 positive cell on the surface of K562 cells were measured by Flow Cytometry.The adherent rate between K562 and RF/6A was measured by the rose Bengal staining test.Results The percentages of CD18 positive cell in the group A,B,C and D were significantly different (F=14.33,P=0.01).The percentage of group B was significantly higher than that in group A (P=0.001) ; the percentage of group C was significantly lower than that in group B (P=0.001) and group D (P=0.02) ; the difference between group C and A was not significant (95％CI=-14.89-2.13,P=0.12).The differences of the percentage of ninjurin-1 positive cell among the group A,B,C and D were significant (F=39.38,P=0.001).The percentage of group B was significantly higher than that in group A (P=0.00); the difference of the percentage between group C and B was not significant (P =0.06),that was also not significant between group C and D (P=0.49).The differences of the adherent rate between K562 and RF/6A (rhesus monkey retinal choroid blood vessel endothelial cell line) among the group A,B,C and D were significant (F=20.62,P=0.00).The adherent rate of group B was significantly higher than that in group A (P=0.00),the adherent rate in group C was significantly lower than that in group B (P=0.01),but it was still significantly higher than that in group A (P=0.002),the difference of adherent rate between group B and D was not significant (P=0.68).Conclusion Under the early stage of diabetic retinopathy,HIF-1α small interfere RNA pSUPERH1-siHIF1α may significantly suppress the expression of CD18 on the surface of K562 cells,but it may not significantly influence the expression of ninjurin-1 on the surface of K562 cells.
