摘要
目的应用血清药理学方法探讨补肾活血中药对体外培养的视网膜Müller细胞在缺氧及转化生长因子-β2(transforming growth factorβ2,TGF-β2)干预下谷氨酸(glutamate,Glu)代谢的影响。方法取新生7 d SD大鼠5只,颈椎脱位处死后进行视网膜Müller细胞培养。根据[3H]标记的D,L-Glu摄入量判断体外培养第2代视网膜Müller细胞的Glu的摄入功能;以酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)测定体外培养细胞外液中谷氨酰胺合成酶(glutamine synthetase,GS)活性。结果与正常对照组比较,12 h、24 h、48 h时缺氧组、TGF-β2+缺氧组的Glu摄入量增加(均为P<0.05),而24 h、48h时GS活性明显下降(均为P<0.05);与缺氧组比较,TGF-β2+缺氧组12 h、48 h时Glu摄入量减少,48 h时GS活性下降(P<0.05)。24 h、48 h时TGF-β2干预组较正常对照组Glu摄入量增加(均为P<0.05),12 h时GS活性增加(P<0.05)。与TGF-β2+缺氧组比较,中药血清+TGF-β2+缺氧组12 h、24 h时Glu摄入量减少(均为P<0.05),但GS的活性增加(均为P<0.05)。结论缺氧使视网膜Müller细胞的Glu摄入功能明显增强,GS活性明显降低,而TGF-β2加重缺氧条件下Müller细胞GS活性的降低;在缺氧和TGF-β2同时存在条件下,补肾活血中药血清下调视网膜Müller细胞的Glu摄入能力,增加GS的活性。
Objeetive To explore the effect of bushenhuoxue decoction on Gin metabolism in transforming growth factor-β2 （ TGFβ2 ）-intervened retinal muller cells cultured in vitro under hypoxic condition using serum pharmacological methods. Methods Five 7-day-old SD rats were selected and executed using cervical disloca- tion,and then the retinal miiller cells were cultured in vitro. Glu uptake capability of the second-generation mtiller cells was determined by measuring the uptake of [^3H]-la- beled D,L-Ghi. Glutamine synthetase（GS） activity in the extracellular fluid was meas- ured by enzyme-linked immunosorbent assay （ELISA）. Results Compared with the normal control group, hypoxia group and TGF-β2 ＋ hypoxia group had higher Gin uptake at the 12th,24th and 48th hour（ All P 〈 0.05 ） ;While much lower GS activity at the 24th and 48th hour（ All P 〈 0.05 ）. Compared with hypoxia group, TGF-β2 ＋ hypoxia group had higher Glu uptake at the 12th and 48th hour（ Both P 〈 0.05 ） ; While a lower GS ac- tivity at the 48th hour（P 〈 0.05 ）. Compared with the normal control group, TGF-β2-in- tervened group had lower Gin uptake at the 24th and 48th hour （ Both P 〈 0.05 ）, yet higher GS activity at the the 12th hour（P 〈 0.05 ）. Compared with TGF-β2 ＋ hypoxia group,bushenhuoxue decoction ＋ TGF-β2 ＋ hypoxia group had lower Glu uptake （Both P〈0. 05） ,yet higher GS activity at the 12th and 24th hour（ Both P 〈 0.05 ）. Conelu- sion Hypoxia could induce an great increase of Glu uptake capability and a significant decrease of GS activity of retinal mttUer cells. TGF-β2 intervention could aggravate the hypoxia-induced decrease of GS activity of retinal mttller cells. Serum with bushenhuox- ue decoction could down-regulate the Ghi uptake capability and enhance GS activity of TGF-β2-intervened retinal mtiller cells under hypoxic condition.
出处
《眼科新进展》
CAS
北大核心
2013年第9期804-807,共4页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:30572407)~~
作者简介
马殿伟，男，1975年4月出生，博士，主治医师。联系电话：13308194001；E—mail：dr—madianwei@163．com
通讯作者：谢学军，E-mail：xxj8848@163．com
