中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2011年
2期
160-162
,共3页
细胞黏附分子/拮抗剂和抑制剂%透明质酸/代谢%Müller细胞
細胞黏附分子/拮抗劑和抑製劑%透明質痠/代謝%Müller細胞
세포점부분자/길항제화억제제%투명질산/대사%Müller세포
Cell adhesion molecules/antagonists & inhibitors%Hyaluronic acid/metabolism%Müller cells
目的 观察CD44抗体对视网膜Müller细胞降解透明质酸(HA)功能的影响.方法 培养的猪后极部视网膜Müller细胞,取第2代细胞用于实验.细胞分为1、2、3组,每组含12×103个Müller细胞.其中1组培养液中未加入其他试剂;2组培养液中加入0.01 mg/ml HA;3组培养液中加入0.01 mg/ml HA和10μg/ml抗CD44抗体.2、3组细胞和HA、单克隆抗CD44抗体预培养,收集1、2、3组上清液,行HA-底物胶电泳法和类酶链免疫吸附测定(ELISA)法检测.结果 HA-底物胶电泳法分析结果显示,1组表现为蓝色背景下的白色细淡的双条带;2组双条带明显增厚,合并成较粗和明亮的脱色条块;3组脱色条块回复为细淡的双条带.类ELISA法检测结果显示,1、2、3组吸光度[A,旧称光密度(OD)]值分别为0.310±0.025、0.093±0.051、0.025±0.069.2组A值较1组A值明显降低,与1组A值比较,差异有统计学意义(t=28.1,P＜0.01);3组A值与1组A值比较,差异无统计学意义(t=4.92,P＞0.05),与2组A值比较,差异有统计学意义(t=26.9,P＜0.01).结论 Müller细胞与HA的相互作用可加强细胞降解HA的功能,CD44抗体可降低这种加强作用.
目的 觀察CD44抗體對視網膜Müller細胞降解透明質痠(HA)功能的影響.方法 培養的豬後極部視網膜Müller細胞,取第2代細胞用于實驗.細胞分為1、2、3組,每組含12×103箇Müller細胞.其中1組培養液中未加入其他試劑;2組培養液中加入0.01 mg/ml HA;3組培養液中加入0.01 mg/ml HA和10μg/ml抗CD44抗體.2、3組細胞和HA、單剋隆抗CD44抗體預培養,收集1、2、3組上清液,行HA-底物膠電泳法和類酶鏈免疫吸附測定(ELISA)法檢測.結果 HA-底物膠電泳法分析結果顯示,1組錶現為藍色揹景下的白色細淡的雙條帶;2組雙條帶明顯增厚,閤併成較粗和明亮的脫色條塊;3組脫色條塊迴複為細淡的雙條帶.類ELISA法檢測結果顯示,1、2、3組吸光度[A,舊稱光密度(OD)]值分彆為0.310±0.025、0.093±0.051、0.025±0.069.2組A值較1組A值明顯降低,與1組A值比較,差異有統計學意義(t=28.1,P＜0.01);3組A值與1組A值比較,差異無統計學意義(t=4.92,P＞0.05),與2組A值比較,差異有統計學意義(t=26.9,P＜0.01).結論 Müller細胞與HA的相互作用可加彊細胞降解HA的功能,CD44抗體可降低這種加彊作用.
목적 관찰CD44항체대시망막Müller세포강해투명질산(HA)공능적영향.방법 배양적저후겁부시망막Müller세포,취제2대세포용우실험.세포분위1、2、3조,매조함12×103개Müller세포.기중1조배양액중미가입기타시제;2조배양액중가입0.01 mg/ml HA;3조배양액중가입0.01 mg/ml HA화10μg/ml항CD44항체.2、3조세포화HA、단극륭항CD44항체예배양,수집1、2、3조상청액,행HA-저물효전영법화류매련면역흡부측정(ELISA)법검측.결과 HA-저물효전영법분석결과현시,1조표현위람색배경하적백색세담적쌍조대;2조쌍조대명현증후,합병성교조화명량적탈색조괴;3조탈색조괴회복위세담적쌍조대.류ELISA법검측결과현시,1、2、3조흡광도[A,구칭광밀도(OD)]치분별위0.310±0.025、0.093±0.051、0.025±0.069.2조A치교1조A치명현강저,여1조A치비교,차이유통계학의의(t=28.1,P＜0.01);3조A치여1조A치비교,차이무통계학의의(t=4.92,P＞0.05),여2조A치비교,차이유통계학의의(t=26.9,P＜0.01).결론 Müller세포여HA적상호작용가가강세포강해HA적공능,CD44항체가강저저충가강작용.
Objective To observe the effect of CD44 antibody on the hyaluronic acid (HA)generation) were cultured in three different medium: without HA (group 1),0. 01 mg/ml HA (group 2),10 μg/ml HA and CD44 antibody (group 3). The cells in the group 2 and 3 were pre-cultured with HA and CD44 antibody, and the supernatant was collected. HA-substrate gel electrophoresis was performed for HA degradation, while ELISA-like method was performed for HA-binding protein. Results HA-substrate gel electrophoresis showed white light double-band on blue background in groups 1 and 3, thicker double-band or bright de-colored blocks in group 2. ELISA-like method showed that the absorbance (A) value of groups 1,2 and 3 were 0.310 ± 0.025, 0.093 ± 0.051, 0.025 ± 0.069 respectively. The A value of group 2 was obviously lower than that of group 1 (t=28.1, P＜0.01); the A value of group 3 was significantly higher than that of group 2 (t=26.9, P＜0.01), but was the same as group 1 (t=4.92, P＞0.05). Conclusion CD44 antibody can inhibit the interaction between Miller cells and HA, and thus reduce the HA degradation.
