中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2014年
6期
594-598
,共5页
马维%汪晓磊%章欣欣%陈璐勔%韩松%李俊发
馬維%汪曉磊%章訢訢%陳璐勔%韓鬆%李俊髮
마유%왕효뢰%장흔흔%진로면%한송%리준발
脉络膜新生血管化/病理生理学%转化生长因子β/激动剂%肿瘤坏死因子α%血管内皮生长因子类%动物实验
脈絡膜新生血管化/病理生理學%轉化生長因子β/激動劑%腫瘤壞死因子α%血管內皮生長因子類%動物實驗
맥락막신생혈관화/병리생이학%전화생장인자β/격동제%종류배사인자α%혈관내피생장인자류%동물실험
Choroidal neovascularization/physiopathology%Transforming growth factor beta/ agonists%Tumor necrosis factor-alpha%Vascular endothelial growth factors%Animal experimentation
目的 观察转化生长因子-β(TGF-β)在激光诱导脉络膜新生血管形成(CNV)中的作用.方法 6～8周龄雄性C57BL/6J小鼠80只随机分为正常对照组、光凝模型组、光凝加磷酸盐缓冲液(PBS)对照组(PBS对照组)、光凝加TGF-β受体抑制剂组(TGF-β受体抑制剂组),每组20只.光凝模型组、PBS对照组及TGF-β受体抑制剂组小鼠均采用激光光凝诱导CNV形成.激光光凝后1、2、3、4周,对正常对照组及光凝模型组小鼠行荧光素眼底血管造影(FFA)检查,动态观察小鼠模型CNV的形成情况;采用蛋白免疫印迹法(Western blot)检测正常对照组及光凝模型组小鼠视网膜内TGF-β的蛋白表达以及正常对照组、PBS对照组、TGF-β受体抑制剂组小鼠视网膜内VEGF和TNF-α的蛋白表达.激光光凝后2周,作视网膜色素上皮细胞/脉络膜组织铺片,采用荧光染色计算各组CNV面积.结果 FFA检查结果显示,正常对照组小鼠视网膜血管以视盘为中心呈放射状走行,脉络膜血管呈网状分布;激光光凝后1周,光凝模型组小鼠光凝区可见荧光素渗漏呈盘状强荧光团.Western blot检测结果显示,光凝模型组小鼠视网膜内TGF-β蛋白表达随激光光凝时间延长而逐渐升高.光凝模型组与正常对照组小鼠视网膜内TGF-β蛋白表达比较,差异有统计学意义(F=13.042,P＜0.05).激光光凝后1、2、3、4周,PBS对照组、TGF-β受体抑制剂组小鼠视网膜内TNF-α(F=14.721、17.509)、VEGF(F=18.890、11.251)蛋白表达均较正常对照组明显增加,差异均有统计学意义(P＜0.05).与PBS对照组比较,TGF-β受体抑制剂组小鼠视网膜内TNF-α、VEGF蛋白表达明显降低,差异均有统计学意义(F=21.321、16.160,P＜0.05).激光光凝后2周,TGF-β受体抑制剂组小鼠脉络膜内CNV面积明显小于PBS对照组及光凝模型组,差异均有统计学意义(F=4.482,P＜0.05).结论 TGF-β可能通过上调TNF-α和VEGF蛋白表达水平促进CNV形成,应用其抑制剂可以减少CNV形成.
目的 觀察轉化生長因子-β(TGF-β)在激光誘導脈絡膜新生血管形成(CNV)中的作用.方法 6～8週齡雄性C57BL/6J小鼠80隻隨機分為正常對照組、光凝模型組、光凝加燐痠鹽緩遲液(PBS)對照組(PBS對照組)、光凝加TGF-β受體抑製劑組(TGF-β受體抑製劑組),每組20隻.光凝模型組、PBS對照組及TGF-β受體抑製劑組小鼠均採用激光光凝誘導CNV形成.激光光凝後1、2、3、4週,對正常對照組及光凝模型組小鼠行熒光素眼底血管造影(FFA)檢查,動態觀察小鼠模型CNV的形成情況;採用蛋白免疫印跡法(Western blot)檢測正常對照組及光凝模型組小鼠視網膜內TGF-β的蛋白錶達以及正常對照組、PBS對照組、TGF-β受體抑製劑組小鼠視網膜內VEGF和TNF-α的蛋白錶達.激光光凝後2週,作視網膜色素上皮細胞/脈絡膜組織鋪片,採用熒光染色計算各組CNV麵積.結果 FFA檢查結果顯示,正常對照組小鼠視網膜血管以視盤為中心呈放射狀走行,脈絡膜血管呈網狀分佈;激光光凝後1週,光凝模型組小鼠光凝區可見熒光素滲漏呈盤狀彊熒光糰.Western blot檢測結果顯示,光凝模型組小鼠視網膜內TGF-β蛋白錶達隨激光光凝時間延長而逐漸升高.光凝模型組與正常對照組小鼠視網膜內TGF-β蛋白錶達比較,差異有統計學意義(F=13.042,P＜0.05).激光光凝後1、2、3、4週,PBS對照組、TGF-β受體抑製劑組小鼠視網膜內TNF-α(F=14.721、17.509)、VEGF(F=18.890、11.251)蛋白錶達均較正常對照組明顯增加,差異均有統計學意義(P＜0.05).與PBS對照組比較,TGF-β受體抑製劑組小鼠視網膜內TNF-α、VEGF蛋白錶達明顯降低,差異均有統計學意義(F=21.321、16.160,P＜0.05).激光光凝後2週,TGF-β受體抑製劑組小鼠脈絡膜內CNV麵積明顯小于PBS對照組及光凝模型組,差異均有統計學意義(F=4.482,P＜0.05).結論 TGF-β可能通過上調TNF-α和VEGF蛋白錶達水平促進CNV形成,應用其抑製劑可以減少CNV形成.
목적 관찰전화생장인자-β(TGF-β)재격광유도맥락막신생혈관형성(CNV)중적작용.방법 6～8주령웅성C57BL/6J소서80지수궤분위정상대조조、광응모형조、광응가린산염완충액(PBS)대조조(PBS대조조)、광응가TGF-β수체억제제조(TGF-β수체억제제조),매조20지.광응모형조、PBS대조조급TGF-β수체억제제조소서균채용격광광응유도CNV형성.격광광응후1、2、3、4주,대정상대조조급광응모형조소서행형광소안저혈관조영(FFA)검사,동태관찰소서모형CNV적형성정황;채용단백면역인적법(Western blot)검측정상대조조급광응모형조소서시망막내TGF-β적단백표체이급정상대조조、PBS대조조、TGF-β수체억제제조소서시망막내VEGF화TNF-α적단백표체.격광광응후2주,작시망막색소상피세포/맥락막조직포편,채용형광염색계산각조CNV면적.결과 FFA검사결과현시,정상대조조소서시망막혈관이시반위중심정방사상주행,맥락막혈관정망상분포;격광광응후1주,광응모형조소서광응구가견형광소삼루정반상강형광단.Western blot검측결과현시,광응모형조소서시망막내TGF-β단백표체수격광광응시간연장이축점승고.광응모형조여정상대조조소서시망막내TGF-β단백표체비교,차이유통계학의의(F=13.042,P＜0.05).격광광응후1、2、3、4주,PBS대조조、TGF-β수체억제제조소서시망막내TNF-α(F=14.721、17.509)、VEGF(F=18.890、11.251)단백표체균교정상대조조명현증가,차이균유통계학의의(P＜0.05).여PBS대조조비교,TGF-β수체억제제조소서시망막내TNF-α、VEGF단백표체명현강저,차이균유통계학의의(F=21.321、16.160,P＜0.05).격광광응후2주,TGF-β수체억제제조소서맥락막내CNV면적명현소우PBS대조조급광응모형조,차이균유통계학의의(F=4.482,P＜0.05).결론 TGF-β가능통과상조TNF-α화VEGF단백표체수평촉진CNV형성,응용기억제제가이감소CNV형성.
Objective To investigate the effects of transforming growth factor-β (TGF-β) in choroidal neovascularization (CNV) induced by laser in mice.Methods Eighty male C57BL/6J mice at the age of 6-8 weeks old were randomly divided into the normal control,photocoagulation model,photocoagulation with phosphate buffered saline (PBS control group) and photocoagulation with TGF-β receptor inhibitor groups (TGF-β receptor inhibitor group),twenty mice of each group.Fundus argon laser photocoagulation was performed in the photocoagulation model group,PBS control group and TGF-β receptor inhibitor group to induce CNV.One week,two,three and four weeks after the laser procedure,fundus fluorescein angiography (FFA) was carried out in the normal control or photocoagulation model groups to observe CNV formation dynamically.Western blot was used to analyze the expressions of TGF-β in the retina from the mice of normal control or photocoagulation model groups,and VEGF or TNF-α in the retina of normal control,PBS control or TGF-β receptor inhibitor groups.The CNV areas of each group were evaluated by using fluorescein stain on retinal pigment epithelium (RPE)/choroid flat mounts after two weeks of photocoagulation.Results The FFA results showed the retinal vessels centered on the optic disc and arranged radially,while the choroidal vascular present network distribution in the normal control mice.Significant leakage of fluorescein showed discoid strong fluorophore in photocoagulation sites of retina at one week after photocoagulation.The quantitative analysis results of Western blot demonstrated that the TGF-β protein expression levels in retina of photocoagulation model mice gradually increased with time passing.The protein expression levels of TGF-β were significant differences in the photocoagulation model group comparing with the normal control group (F=13.042,P＜0.05).The protein expression levels of TNF-α (F=14.721,17.509) and VEGF (F=18.890,11.251) increased significantly in retina of PBS control or TGF-β receptor inhibitor groups when compared with that of normal control group at one week,two,three and four weeks after photocoagulation,and the differences were both statistically significant (P＜0.05).Compared with PBS control group,the protein levels of TNF-α and VEGF in retina from TGF-β receptor inhibitor group were significantly reduced,the differences was statistically significant (F=21.321,16.160,P＜0.05).Two weeks after laser photocoagulation,a distinct reduction in CNV lesion size in the TGF-β receptor inhibitor group mice when compared to PBS or normal control groups,the differences was statically significant (F=4.482,P＜0.05).Conclusion TGF β may promote CNV formation by up-regulating both TNF α and VEGF protein expressions,the application of its specific inhibitor is able to reduce CNV progression.
