染料木黄酮对兔结膜成纤维细胞增殖的抑制作用

目的:观察染料木黄酮(genistein,Geni)对体外培养的兔结膜成纤维细胞(rabbitconjunctivalfibroblasts,RCF)增殖的影响。方法:应用MTT法测定不同生长时间及浓度的Geni,5-FU及二者联合药物对RCF生长的影响。结果:2.5￣40mg/LGeni对RCF生长抑制率分别为27.2%,34.0%,44.6%,55.1%,58.9%;不同浓度的Geni作用1,3,5,7d的生长抑制率,随时间、剂量增加而加大。Geni(5,10mg/L)与5-FU(1mg/L)联用的抑制率分别为45.6%和49.1%。结论:Geni对RCF的增殖有抑制作用,呈时间和剂量依赖性;与5-FU有协同效应。     

Ge—132对兔结膜成纤维细胞增殖的影响

用ＭＴＴ比色法和细胞计数法测定细胞增殖。观察了Ｇｅ－１３２（羧乙基锗倍半氧化物）在体外对兔结膜成纤维细胞增殖的影响。结果显示：Ｇｅ－１３２药物最终浓度在０．００１６—５ｍｇ／ｍｌ范围有剂量依从性的抑制成纤维细胞增殖的作用，其半数有效浓度（ＩＣ５０）为０．１４７ｍｇ／ｍｌ。它有可能用于控制成纤维细胞的过度增殖。     

汉防己甲素及地塞米松对兔结膜成纤维细胞增殖的抑制作用

目的　观察汉防己甲素 (tetrandrine,Tet)对体外培养的兔结膜成纤维细胞 (rab-bit conjunctival fibroblasts,RCF)增殖的抑制作用 ,探讨其与地塞米松 (dexamethasone,Dex)联合应用的价值 ,并对二者的抑制作用进行比较。方法　应用 MTT法 ,测定不同浓度的 Tet、Dex及 Tet联合 Dex对 RCF增殖的抑制作用。结果　浓度为 1 0 0、80、2 0、5、1 .2 5、0 .3 1 2 5mg· L-1的 Tet对 RCF的增殖均有抑制作用 ,其抑制率分别为 86.0 0 %、78.1 5%、62 .89%、3 0 .1 9%、1 8.0 4%、9.2 1 %,与对照组相比有显著性差异。半数抑制量(ID50 ) 2 0 0 mg·L-1的 Dex在联合 5mg· L-1的 Tet后 ,对细胞生长的抑制率为 49.2 5%.结论　 Tet对 RCF的增殖有抑制作用 ,且与药物浓度成正相关。 Dex联合 Tet有协同抑制作用。 Tet可能成为一种新型的抗眼部增殖性疾病的药物     

MTT法检测紫杉醇对兔晶状体上皮细胞增殖的影响

目的　研究紫杉醇 (Taxol)对体外培养的兔晶状体上皮细胞生长的影响。方法　取 2～ 3代对数生长期的兔晶状体上皮细胞 ,于 96孔板中培养 2 4 h后 ,用不同浓度的紫杉醇分别作用 2 4及 72 h,MTT比色测定法观察紫杉醇对兔晶状体上皮细胞增殖的影响。结果　我们发现 ,浓度为 0 .312 5 m g· L- 1的紫杉醇作用于培养的兔晶状体上皮细胞 2 4 h对其生长无明显的抑制 ,而作用 72 h则能抑制其生长。 2 4 h的半效抑制量 (ID50 )为 14 .6 81mg· L- 1 ,72 h的 ID50 为 4 .0 0 7mg· L- 1 ,紫杉醇还能抑制细胞贴壁。结论　紫杉醇对传代培养的兔晶状体上皮细胞增殖有明显的抑制作用 ,紫杉醇对预防后囊膜混浊的发生可能有一定作用     

几丁聚糖对兔结膜上皮细胞和结膜成纤维细胞增殖的影响

目的　研究几丁聚糖对兔结膜上皮细胞和结膜成纤维细胞增殖的影响。方法　采用消化法培养结膜上皮细胞 ,组织块法培养结膜成纤维细胞。通过MTT比色法和细胞计数法测定几丁聚糖对两种细胞增殖的影响。结果　几丁聚糖在浓度 >0 0 3 %时能促进结膜上皮细胞的增殖 ,浓度为 0 12 %时促进作用最明显。结膜成纤维细胞在几丁聚糖浓度 >0 0 3 %时其增殖即受到抑制 ,抑制作用呈浓度依赖性。结论　几丁聚糖有望用于预防睑球粘连     

肝素单独应用及与红比霉素、阿霉素联合应用抑制结膜成纤维细胞增殖

我们测定了肝素单独应用及肝素分别与红比霉素、阿霉素联合应用在体外对兔结膜成纤维细胞的影响。证实肝素单独应用可明显抑制结膜成纤维细胞增殖,肝素的ID_(50)值为1.93mg/ml,肝素单独应用抑制成纤维细胞生长的效力比其与红比霉素或阿霉素联合应用的效力更大。肝素无明显副作用,有希望成为提高青光眼手术成功率的理想药物。     

MTT法探讨三氧化二砷及莪术对晶状体上皮细胞增殖的抑制作用

目的探讨两种天然药物三氧化二砷(ATO)和莪术(RC)对晶状体上皮细胞(LEC)增殖的抑制作用,为寻求防治后发性白内障的确切有效药物提供实验依据。设计实验性研究。研究对象体外培养的牛LEC。方法倒置显微镜下观察细胞形态学变化。MTT比色法检测不同终浓度(2．5、5、10μmol/L)ATO和(5、10、20mg/ml)RC作用于增殖状态下LEC 72小时后对细胞增殖的抑制程度。主要指标细胞形态、吸光度(A)值。结果倒置显微镜下可见增殖对照组细胞大小均匀,生长密集,ATO和RC组细胞生长及形态有明显改变,增殖缓慢,生长差,贴壁能力减弱。MTT比色法结果显示:不同浓度(2．5、5、 10μmol／L)的ATO和(5、10、20mg／ml)RC可明显抑制LEC增殖,并具有剂量依赖关系(P均<0．01)。结论两种天然药物 ATO、RC能够明显抑制LEC增殖,有望成为防治后发性白内障的有效药物。     

用MTT比色法测定兔眼结膜成纤维细胞的增殖与存活

对Mosmann的MTT比色方法进行改良后,应用于测定兔眼结膜成纤维细胞的体外增殖与存活,获得满意结果。此方法具有快速、简便、精确、不需任何同位素、不受主观因素影响等优点,可代替~3H-Tdr掺入法测定成纤维细胞及其它细胞增殖,用于药物筛选等研究,值得推广。     

含不同分子量蛋白质的视网膜下液促成纤维细胞增殖的实验研究

含不同分子量蛋白质的视网膜下液促成纤维细胞增殖的实验研究唐罗生姜德咏聂爱光罗贤明朱晓华为探讨视网膜下液（ｓｕｂｒｅｔｉｎａｌｆｌｕｉｄ，ＳＲＦ）中具有生长因子样作用蛋白质与增殖性玻璃体视网膜病变（ｐｒｏｌｉｆｅｒａｔｉｖｅｖｉｔｒｏｒｅｔｉｎｏｐａｔ...     

血小板源性生长因子BB对体外培养兔角膜成纤维细胞增殖和细胞周期的影响

目的探讨重组人血小板源性生长因子BB(recombinant human platelet-derived growth factor-BB,rhPDGF-BB)对体外培养的兔角膜成纤维细胞增殖和细胞周期的影响。方法体外培养兔角膜成纤维细胞,分实验组和对照组,对照组弃原培养液,加无血清培养液继续培养,实验组用不同浓度的rhPDGF-BB1μg·L-1、10μg·L-1、100μg·L-1、1000μg·L-1分别作用于兔角膜成纤维细胞24h、48h、72h,用MTT比色法检测rhPDGFBB对兔角膜成纤维细胞生长的影响,流式细胞仪检测rhPDGFBB对兔角膜成纤维细胞细胞周期的影响。结果rhPDGFBB在1～100μg·L-1浓度范围内与兔角膜成纤维细胞增殖呈剂量效应关系,当浓度增大到1000μg·L-1,促增殖作用减弱。流式细胞术对细胞周期时项分析结果显示,经rhPDGFBB作用后,兔角膜成纤维细胞的DNA合成量(S%)显著升高,G1%降低,反映细胞增殖活力的增殖指数PI值增高。结论rhPDGFBB能促进兔角膜成纤维细胞的增殖和DNA合成,这一作用可能是通过促使处于G1期的兔角膜成纤维细胞进入S期来实现的。     

低分子量肝素防治兔增生性玻璃体视网膜病变的实验研究

目的 :探讨低分子量肝素 (lowmolecularweightheparin ,LMWH)对因玻璃体出血所致的增生性玻璃体视网膜病变(proliferativevitreoretinopathy ,PVR)的防治作用 ,并研究其与临床上常用的与防治PVR有关的药物地塞米松联合应用的效果。方法 :2 0只健康有色家兔 ,随机分成 4组 ,即对照组、LMWH组 (10 0IU)、地塞米松组 (5 0 0 μg)及LMWH(10 0IU) +地塞米松 (5 0 0 μg)组 (均以生理盐水稀释成 0 2ml) ,每组 5只 ,计 10只眼。采用经睫状体玻璃体腔注入药物及自体新鲜血 0 1ml的方法 ,观察LMWH对实验性兔玻璃体视网膜病变的影响 ,并与对照组、地塞米松、及联合用药组进行比较。结果 :术后 14、2 1及 2 8天各组的平均增生级别分别为 :对照组 ,2 6、2 7和 3 4;LMWH组 ,0 75、1 0和 1 2 ;地塞米松组 ,2 6、2 7和 3 3;LMWH联合地塞米松组 ,0 6、0 6 5和 1 0 5。各时期LMWH组及LMWH联合地塞米松组与对照组及地塞米松组均有显著性差异 (P <0 0 5 ) ;LMWH组与联合用药组无显著性差异 (P >0 0 5 ) ;地塞米松组与对照组无显著性差异 (P >0 0 5 )。结论 :LMWH对经兔睫状体玻璃体腔注入新鲜自体血继发的PVR有防治作用 ,地塞米松未见有防治作用 ,LMWH、地塞米松联合应用亦未见明显的协同作用     

冬凌草甲素抑制兔小梁切除术后培养的结膜成纤维细胞增殖的初步研究

目的观察冬凌草甲素（Ori）对小梁切除术后培养的免结膜成纤维细胞（RCF）增殖的影响。设计实验研究。研究对象体外培养的小梁切除术后结膜滤过区来源RCF。方法制作兔眼小梁切除术模型,术后7天取兔滤过区结膜下组织进行成纤维细胞体外培养。分别用浓度依次为5、10、20、30、40、50μmol·L^-1。的Ori干预48小时。采用MTT法检测不同浓度药物对成纤维细胞生长的影响。主要指标成纤维细胞的光密度（OD）值和生长抑制率。结果加入Ori培养48小时后,5、10、20、30、40、50μmol·L^-1不同浓度组成纤维细胞的OD值分别为（0．402±0．013）、（0．347±0．011）、（0．293±0．015）、（0．236±0．022）、（0．184±0．023）和（O．125±0．014）,与无药对照组OD值（0．453±0．015）比较差异均有统计学意义（P=0．021～0．000）,并呈浓度依赖性减小（F=5．48,P=0．003）。浓度为5、10、20、30、40、50μmol·L^-1的Ori对成纤维细胞生长抑制率分别为11．26％、23．40％、35．32％、47．90％、59．38％和72．41％。结论Ori可以抑制培养的兔眼滤过区结膜成纤维细胞的增殖,抑制作用呈剂量依赖性。Ori可能成为一种治疗青光眼术后滤过泡纤维化的新药物。（眼科,2009,18：128-130）     

低相对分子质量肝素对牛晶状体上皮细胞在人工晶状体表面增殖的影响

目的　体外比较研究低相对分子质量肝素 (lowmolecularweightheparin ,LMWH)和普通肝素影响牛晶状体上皮细胞 (bovinelensepithelialcell,BLEC)在聚甲基丙烯酸甲酯 (polymethylmethacrylate,PMMA)人工晶状体表面增殖的作用。方法　体外将BLEC接种到PMMA人工晶状体表面培养 ,LMWH组和肝素组培养液中分别含LMWH(1× 10 5U/L)和普通肝素 (1× 10 5U/L) ,对照组培养液中不含上述药物。 72h后用流式细胞仪对人工晶状体表面黏附的细胞进行计数分析。结果　经LMWH和普通肝素处理的PMMA人工晶状体表面BLEC形态幼稚 ,对照组BLEC呈纤维化趋势 ;细胞计数对照组为(878 2± 10 7 8)个 /ml,肝素组为 (5 4 5 5± 5 8 2 )个 /ml,LMWH组为 (40 7 8± 36 6 )个 /ml,3个组比较差异有非常显著意义 (P <0 0 0 1)。结论　LMWH可显著抑制晶状体上皮细胞在人工晶状体表面的黏附和增殖 ,有望成为更为安全、有效的防治后发性白内障的药物     

低分子肝素缓释系统防治兔后发性白内障的实验研究

目的 探讨低分子肝素缓释系统防治兔后发性白内障的有效性和安全性.方法 采用前瞻性随机分组对照研究.以乳酸-羟基乙酸共聚物(PLGA)为载体采用冻干法制备低分子肝素缓释系统(LMWH DDS)并体外评价其缓释特性.将50只(50只眼)新西兰白兔分别行超声乳化透明晶状体吸除术,并随机均分为5组:A组术后生理盐水滴眼,B、C、D组术毕后房分别植入载药量为1.00 mg、0.50 mg、0.25 mg的LMWH DDS,E组植入不含药物的空白缓释系统;术后12周对术眼行裂隙灯显微镜、组织病理学以及电镜检查,并检测房水药物浓度和晶状体后囊膜湿重.术后房水闪光、房水细胞分级以及后囊膜混浊分级资料采用Kruskal-Wallis检验,房水药物浓度采用具有一个重复测量因素的两因素方差分析.结果 采用冻干法制备的LMWH DDS包封率为98.2%,体外释药方程拟合以零级方程为佳.术后B、C、D组炎症反应较A、E组显著减轻;术后12周A、B、C、D、E各组后囊膜混浊发生率分别为100%(10/10)、20%(2/10)、30%(3/10)、90%(9/10)、100%(10/10),后囊膜混浊分级评分组间比较差异均具有统计学意义(X~2=31.637,P=0.000),后囊膜湿重分别为(114.59±14.58)mg、(24.14±6.08)mg、(39.23±17.13)mg、(99.35±29.37)rag、(115.29±19.87)mg,组间比较差异均具有统计学意义(F=42.149,P=0.000);术后4周内B、C组房水中低分子肝素一直维持较高浓度(大于20 ms/L),D组浓度较低且不稳定;光镜和电镜下B、C组后囊细胞增生不活跃,未发现眼内毒性反应;术后均未见眼内出血现象.结论 以PLGA为载体采用冻干法制备的LMWHDDS具有良好的缓释性和组织相容性;其后房植入能明显减轻术后炎症反应,能安全、有效抑制后发性白内障的发生,且存在一定量效关系.     

Histopathologic effects of a low molecular weight heparin on bone healing in rats: a promising adjuvant in dacryocystorhinostomy

AIM: To investigate the effect of short-term prophylactic dose of a low molecular weight heparin (LMWH) drug on the bone healing process in an animal model simulating the osteotomy obtained in dacryocystorhinostomy. METHODS: Forty male Wistar albino rats were divided into 2 groups. Subcutaneous injections of enoxaparin 1 mg/kg (enoxaparin-treated group) and saline solution (control group) were performed once daily for 4d, beginning on the first preoperative day. The osteotomy was created at the femoral diaphysis in all animals by using a Kirschner wire. Each group was further divided into 2 subgroups depending on the timing of the second operation, 14 or 21d following initial osteotomy. Patent osteotomy area on the second and the third weeks in each group were calculated by using a computer software on digital micrographs. RESULTS: The patent osteotomy areas at the second and the third weeks were significantly larger in the enoxaparin-treated group than those of the control group (P<0.001 for each time-period). In the control group, the patent osteotomy area at the third week of healing was significantly smaller than that of the second week (P=0.003), whereas there was no significant difference between these two measurements in the enoxaparin-treated group (P=0.185). CONCLUSION: Short-term administration of enoxaparin resultes in a significant alteration in bone healing at 14 and 21d after injury. LMWHs can be regarded as promising alternative adjuvants in dacryocystorhinostomy after being evaluated with further clinical and animal studies.     

Evaluation of antiviral agents for adenovirus using the MTT method in vitro

PURPOSE: Currently, there is no effective treatment for adenoviral conjunctivitis. We evaluated the antiviral inhibitory effect of four antiviral agents against adenovirus (ADV) in vitro. METHODS: Viruses used for the experiment were ADV type 4 (ADV 4), type 8 (ADV 8) and type 37 (ADV 37). We examined four antiviral agents, i.e., cidofovir (HPMPC), zalcitabine (ddC), foscarnet (PFA), and acyclovir (ACV). 50% effective concentration (EC50), 50% cytotoxic concentration (CC50) and selectivity index (SI) of compounds were determined for ADV infection in HEp-2 cells using the 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. RESULTS: HPMPC and ddC showed an inhibitory effect against all three ADVs. In particular, ddC showed more potent and selective inhibition against ADV than HPMPC. PFA and ACV were ineffective against ADV. CONCLUSIONS: HPMPC and ddC were inhibitory against ADV 4, ADV 8 and ADV 37 replication in vitro. The MTT method is rapid and simple for the screening of antiviral agents. We think this method is also very useful for the screening of anti-ADV agents.     

Effect of low molecular weight heparin (enoxaparin) on congenital cataract surgery

AIM: To assess the efficacy of intracameral enoxaparin (a low-molecular-weight heparin) infusion, in variable doses on postoperative inflammatory response in congenital cataract surgery.METHODS: It is aprospective, randomized controlled trial. Eighty eyes of 53 children with congenital cataract were enrolled in this study. Every eye had primary posterior capsulorrhexis and intraocular lens (IOL) implantation after lens aspiration. The eyes were divided into 4 equal groups. In group 1 balanced salt solution (BSS) without enoxaparin was used as an irrigation solution. Whereas in group 2, 3 and 4, 40mg, 20mg and 10mg enoxaparin in 500mL BSS was used respectively. The inflammatory response in the anterior chamber was compared among the groups with slit-lamp biomicroscopy.RESULTS: The mean follow-up period was (17.75±3.95) months in group 1, (18.00±5.15) months in group 2, (19.20±5.47) months in group 3 and (18.65±5.16) months in group 4. Mean number of inflammatory cells in the anterior chamber in group 1 was significantly higher than that of group 2, 3, 4 (P<0.001). There was fibrin formation in the anterior chambers of 3 eyes in group 1 and one eye in group 4. There was synechiae formation in 3 eyes of group 1 and one eye of group 4. There was no significant difference among the groups by means of fibrin or synechiae formation (P>0.05). There were IOL precipitates in 4 eyes of group 1 and 2 eyes of group 4. IOL precipitate formation was significantly higher in group 1 than that of group 2 and 3 in which there was no IOL precipitate (P=0.048). There was IOL subluxation in only one eye of group 1, 3 and 4 while no subluxation was observed in group 2 (P>0.05). There was no statistically significant difference detected about IOL subluxation occurance in all 4 groups (P>0.05).CONCLUSION: Complications of cataract surgery in congenital cataract patients associated with postoperative inflammatory response found to be decreased with the use of enoxaparin in intraocular infusion solutions. Furthermore according to our results the anti-inflammatory effect of enoxaparin was dose dependant.