He-Ne激光对兔眼小梁切除术后滤过道成纤维细胞增殖的影响

目的 研究兔眼小粱切除术前辅助使用He-Ne激光照射滤过区对滤过道成纤维细胞增殖情况的影响.方法 随机选取健康新西兰大白兔29只(58眼)用于实验.实验一:将健康新西兰大白兔8只(16眼)随机分为4组,即100 mW·cm-2、150 mW·cm-2、200 mW·cm-2激光手术组及空白对照组,每组各2只(4眼),激光手术组兔眼使用不同功率密度的He-Ne激光照射,空白对照组兔眼不行激光照射.照射部位选择兔眼鼻上象限角膜缘紧邻上直肌处,照射距离10cm,光斑直径4 mm,照射时间10 min,每天1次,连续照射3d.最后一次照射24h后处死动物,取照射区组织采用病理学和免疫组织化学技术.检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA),并使用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminaldeoxynucleotidyl transferase dUTP nick end labeling,TUNEL)观察照射区成纤维细胞增殖和凋亡的病理改变.实验二:将另外21只兔(42眼),采用200 mW·cm-2 He-Ne激光同上法预处理每只兔右眼手术区,最后一次照射24h后实施小梁切除术(即激光联合小粱切除术组),左眼行单纯小粱切除术为单纯手术对照组,分别于手术后7d、14d、28d检测滤过区组织上述指标.结果 实验一:各He-Ne激光联合小梁切除术组与空白对照组均仅检测到少量PCNA阳性细胞,未见明显TUNEL凋亡细胞,差异均无统计学意义(均为P>0.05).实验二:200 mW·cm-2 He-Ne激光联合小梁切除术组与单纯手术对照组PCNA阳性细胞率术后7d最高,14d显著降低,28d降至最低.术后7d、14d,200 mW·cm-2 He-Ne激光联合小梁切除术组PCNA阳性细胞率分别为(19.34±2.35)%、(10.48±3.29)%,均明显低于单纯手术对照组(22.88±3.29)%、(14.35±2.56)%(均为P<0.05);术后28 d,2组PCNA阳性细胞率均较低,差异无统计学意义(p>0.05).2组各时间点亦未检测到明显TUNEL凋亡细胞.结论 He-Ne激光照射对正常兔眼结膜下及巩膜成纤维细胞活性无明显影响,小粱切除术前辅助使用200 mW·cm-2 He-Ne激光照射可以抑制术后早期成纤维细胞增殖,可能有助于抑制滤过泡的瘢痕化而提高手术成功率.     

CTGF在兔眼小梁切除术后滤过泡瘢痕形成中的表达

目的:以兔为动物模型,观察结缔组织生长因子(connective tissue growth factor,CTGF)在兔眼小梁切除术后滤过泡瘢痕形成过程中的表达,初步探讨CTGF在滤过泡瘢痕形成机制中的作用。方法:健康成年家兔28只,随机选取1眼行小梁切除术,另1眼不手术作为对照,分别于术后第1,3,5,7,10,14,21d分批处死,另取2只兔不手术作为正常对照,于术前处死。摘除眼球,剪取滤过泡区域组织,制成病理切片,行常规HE染色及免疫组织化学检测(SABC法),光镜下观察并行图像定量分析。结果:CTGF在对照组织标本中几乎无阳性表达,手术后表达上调,主要表达于成纤维细胞胞浆中,手术后第5d达高峰。结论:在滤过泡瘢痕形成过程中,CTGF发挥了重要的作用,阻断CTGF的基因表达或抑制其活性,可能为防止青光眼滤过手术的失败提供一个新的治疗途径。     

结膜下注射CTGF抗体对兔青光眼滤过手术后眼压和滤过泡面积的影响

目的:探讨结缔组织生长因子(connective tissue growth factor,CTGF)抗体对兔青光眼滤过手术后滤过泡瘢痕化的抑制作用。方法:家兔5只双眼制作青光眼滤过手术模型。随机选取家兔一眼作为抗体组,分别于手术完成当时和术后5d结膜下注射0.1mL浓度为50mg/L的CTGF抗体;另一眼作为对照组在相同时间点结膜下注射0.1mL磷酸盐缓冲液。术后1,3,5,7,10,14d分别观察滤过泡形态并测量其面积和眼压值。结果:术后7,10和14d抗体组滤过泡面积均大于对照组(P<0.05),眼压均小于对照组(P<0.05)。结论:结膜下注射CTGF抗体可维持兔眼滤过手术后较大的滤过泡面积和较低的眼压。     

兔青光眼术后成纤维细胞内结缔组织生长因子的调节

目的观察青光眼滤过手术后滤过泡瘢痕内成纤维细胞中反义寡核苷酸（ASODN）对结缔组织生长因子（CTGF）的调节,为抗青光眼术后滤过泡区瘢痕形成提出新的解决思路。方法取兔眼小梁切除术后第5天的手术区结膜及板层巩膜行成纤维细胞的原代培养。培养的细胞根据干预方式的不同分为CTGFASODN-脂质体复合物组、CTGFASODN对照组、CTGF随机序列寡核苷酸（SCODN）-脂质体复合物组和空白对照组。利用脂质体介导,将经异硫氰酸荧光素（FITC）标记的CTGFASODN转染至兔抗青光眼滤过手术后滤过泡瘢痕组织成纤维细胞中,利用半定量逆转录-聚合酶链反应（RT-PCR）检测CTGFmRNA在成纤维细胞中的表达,采用Westernblot法检测CTGF蛋白的合成量,以相对灰度值为指标,观察CTGFASODN对CTGF在手术区结膜及板层巩膜行成纤维细胞中表达的抑制效果。结果组织块培养法培养的成纤维细胞在13～14d达到融合。CTGFASODN-脂质体复合物组转染6h后可见培养的细胞质中出现少量黄绿色荧光,12h后荧光依然存在;CTGFmRNA在成纤维细胞中表达的相对灰度值明显低于空白对照组（0.457±0.035vs0.790±0.037）,差异有统计学意义（P〈0.01）;CTGF蛋白水平表达也呈相同趋势（0.177±0.022vs0.557±0.024）（P〈0.01）。CTGFASODN对照组、CTGFSCODN-脂质体复合物组CTGF在成纤维细胞中的表达与空白对照组相比差异均无统计学意义（P〉0.05）。结论脂质体能有效地将CTGFASODN转染进入体外培养的成纤维细胞;CTGF在青光眼滤过手术后的瘢痕形成过程中发挥重要作用;CTGFASODN能在脂质体介导下抑制CTGF在成纤维细胞中的表达。     

外源性结缔组织生长因子在兔眼小梁切除术后滤过泡瘢痕形成中的作用

目的 以兔为动物模型,比较外源性结缔组织生长因子(CTGF)和生理盐水在兔眼小梁切除术后对滤过泡瘢痕形成的影响,初步探讨CTGF在滤过泡瘢痕形成机制中的作用.方法 健康成年家兔8只(16只眼),双眼行小梁切除术,16只眼随机分成2组,每组8只眼,滤过泡内分别注入外源性CTGF和生理盐水,术后观察滤过泡形态及眼压变化.结果 滤过泡内注入CTGF后,滤过泡失败时间为(12.00±0.82)d,与生理盐水组比较,差别有统计学意义(P<0.01);与术前眼压比较,CTGF组和生理盐水组术后在第1、7天,差别均有统计学意义(P<0.01),术后第14、21天,CTGF组差别均无统计学意义(P>0.05),而生理盐水组术后第14天,差别仍有统计学意义(P<0.05);CTGF组眼压术后第7天与第1天相比,差别有统计学意义(P<0.01),而生理盐水组差别无统计学意义(P>0.05);术后第7天,CTGF组眼压与生理盐水组比较,差别有统计学意义(P<0.01).结论 外源性CTGF注入滤过泡内,可以促进滤过泡瘢痕形成,眼压迅速回升,导致手术提前失败,其在滤过泡瘢痕形成过程中发挥了重要作用.     

改良小梁切除术临床疗效观察

0引言常规的小梁切除术是目前最普遍应用的抗青光眼滤过术式,但随着随访时间的延长,滤过手术后眼压失控病例不断增加,手术后2a内的失败率高达15%～30%。其主要原因是手术滤过道瘢痕形成,而再次行滤过手术失败率会更高。因此如何防止巩膜瘘道阻塞保持长时间的功能性滤过作用,已成为小梁切除术研究的难点和热点。我院自2001-10开展改良小梁切除术治疗了几种类型的青光眼15例(17眼),临床观察效果满意,现报告如下。1临床资料在15例青光眼患者中,男5例(5眼),女10例(12眼),年龄41～72岁,其中急性闭角型青光眼14眼,慢性开角型青光眼3眼,术前眼压3.5…     

丁酰化壳聚糖膜在兔眼滤过手术抗瘢痕的研究

目的研究丁酰化壳聚糖膜在兔眼高眼压模型滤过手术中的抗瘢痕作用。方法实验研究。45只青紫蓝兔随机分为3组,每组左眼为实验眼,前房均注射0．3％复方卡波姆溶液（含有0．025％地塞米松）0．3ml,诱导兔眼高眼压模型,3周后眼压为30—40mmHg（1mmHg=0．133kPa）。第1组：单纯小梁切除术;第2组：小梁切除术中使用丝裂霉素c（MMC）;第3组：将丁酰化壳聚糖膜植入小梁切除术的板层巩膜瓣下。右眼均不行手术为对照组。分别于术后1、2、4、8及12周每组随机处死3只兔,制作眼球标本。结果病理切片显示术后2周时,单纯小梁切除组巩膜瓣胶原排列紊乱,炎性细胞浸润较多,伴有新生血管形成。MMC组巩膜瓣胶原排列紊乱,胶原组织短小、断裂,伴有炎性细胞浸润。壳聚糖膜组巩膜瓣间隙可见,胶原排列整齐,炎性细胞较少。术后2周,单纯小梁切除组的眼压〉21mmHg,明显高于MMC组和壳聚糖膜组（F=392．869,P〈0．05）,术后12周。壳聚糖膜组眼压低于单纯小梁切除组和MMC组（F=259．923,P〈0．05）;手术4周后,单纯小梁切除组与MMC组功能性滤过泡比率下降。结论丁酰化壳聚糖膜植入兔眼高眼压模型小梁切除术板层巩膜瓣下,有效抑制纤维细胞增生,维持滤过道通畅,术后并发症少,有助于提高手术成功率。     

生物羊膜在青光眼小梁切除术中的应用研究

目的评价生物羊膜在青光眼小梁切除术中的作用。方法收集2010年1月至2011年6月就诊我院诊断为青光眼患者72例(98只眼),按随机数字表分为两组:小梁切除术组和小梁切除联合羊膜移植术组,由同一位医师完成手术,术前和术后1周,1、3、12及18个月随访观察眼压;18个月时采用超声生物显微镜(UBM)测量滤过泡长径和宽径的长度,计算手术成功率。结果小梁切除联合羊膜移植术组术后眼压的波动较小梁切除术组低。UBM检测发现18个月后小梁切除术组滤过泡长径为(3.81±0.91)mm,小梁切除联合羊膜移植术组滤过泡长径为(7.11±2.08)mm,两者间差异具有统计学意义(t=8.708,P<0.001);小梁切除术组滤过泡宽径为(3.14±0.96)mm,小梁切除联合羊膜移植术组滤过泡宽径为(4.78±1.20)mm,两者间差异具有统计学意义(t=6.324,P<0.001)。18个月时小梁切除术组手术完全成功率为56.8%(21只眼),小梁切除联合羊膜移植术组手术完全成功率为84.4%(27只眼),两者间差异具有统计学意义(χ2=6.182,P=0.013);18个月时小梁切除术组手术条件成功率为75.7%(28只眼),小梁切除联合羊膜移植术组手术条件功率为93.8%(30只眼),两者间差异具有统计学意义(χ2=4.183,P=0.041)。结论小梁切除联合羊膜移植术可明显减轻滤过道瘢痕化,形成功能性滤过泡,提高手术成功率,是青光眼治疗一种安全有效的手术方法。     

羊膜植入在青光眼小梁切除术中的应用

目的 青光眼小梁切除术中羊膜植入防止术后瘢痕性滤过泡形成的作用。方法 对12例15眼原发及继发性青光眼行小梁切除术,术中将羊膜植于巩膜床,并观察术后阻止瘢痕性滤过泡形成的效果及降眼压作用。结果 15眼中,形成功能性滤过泡13眼,无功能滤过泡2眼。术后降眼压效果良好。结论 羊膜植入能有效防止小梁切除术后瘢痕性滤过泡形成,进一步提高抗青光眼手术的成功率。     

贝伐单抗结膜下注射对青光眼滤过术后滤过泡瘢痕化的抑制作用

背景 青光眼滤过手术术后滤过道的瘢痕化是导致手术失败的主要原因.传统的抑制滤过道瘢痕化的方法是丝裂霉素C的应用,但存在较多的并发症.研究表明贝伐单抗具有抑制新生血管和纤维增生的作用,其对青光眼滤过术后滤过泡瘢痕化是否有抑制作用受到关注. 目的 观察贝伐单抗结膜下注射对兔眼小梁切除术后滤过泡纤维瘢痕形成的抑制效果. 方法 按随机数字表法将7～9周龄新西兰大白兔40只随机分为4个组,各组兔右眼均行常规小梁网切除术.贝伐单抗单次注射组兔眼术毕结膜下注射贝伐单抗0.05 ml(25 mg/ml),贝伐单抗多次注射组兔眼分别于术毕及术后3、7d注射贝伐单抗,每次均注射0.05 ml,丝裂霉素C组兔眼术毕局部涂用丝裂霉素C,生理盐水组兔眼术毕以同样的方法注射0.05 ml生理盐水.所有兔眼术后每隔1日用Schi(o)tz眼压计测量眼压,行裂隙灯显微镜检查以观察滤过泡形态及其表面的血管分布,并用卡尺测量和计算滤过泡面积.分别于术后14d和28 d摘取实验眼行滤过泡组织病理学检查,采用免疫组织化学法检测滤过道组织中血管内皮细胞标志物CD31的表达以计算微血管数目. 结果 各组兔眼术后眼压值的总体比较差异无统计学意义(F=0.88,P=0.47).与贝伐单抗单次注射组、丝裂霉素C组和生理盐水组兔眼滤过泡的形态比较,术后7d贝伐单抗多次注射组兔眼滤过泡高度隆起且弥散,表面血管稀疏.贝伐单抗多次注射组兔眼滤过泡生存时间为27 d,而贝伐单抗单次注射组、丝裂霉素C组均为19d,生理盐水组为13d.术后14d各组兔眼滤过道胶原纤维百分比分别为(49.18±1.54)％、(26.41±1.23)％、(50.68±1.87)％和(70.63±1.81)％,贝伐单抗单次注射组、贝伐单抗多次注射组、丝裂霉素C组滤过道胶原纤维百分比均低于生理盐水组,贝伐单抗多次注射组低于贝伐单抗单次注射组,差异均有统计学意义(P＜0.05);术后28 d贝伐单抗多次注射组胶原纤维百分比为(66.82±1.53)％,其他3个组出现瘢痕化.术后14d贝伐单抗多次注射组兔眼滤过道组织中微血管数目明显低于贝伐单抗单次注射组、丝裂霉素C组和生理盐水组,差异均有统计学意义(均P＜0.05).术后28 d贝伐单抗多次注射组兔眼滤过道组织中微血管数且为3.51±0.31,均高于贝伐单抗注射组、丝裂霉素组和生理盐水组,差异均有统计学意义(均P＜0.05). 结论 青光眼滤过术后结膜下注射贝伐单抗有助于维持功能滤过泡的形态,抑制滤过道瘢痕化,提高手术的成功率.     

Influence and mechanism of He-Ne laser on scar formation of filtration canal after trabeculectomy in rabbit

AIM: To investigate the influence of He-Ne laser on connective tissue growth factor (CTGF) expression and collagen formation of fibroblast in filtration site after trabeculectomy in rabbit, and to discuss the mechanism for preventing scar formation with He-Ne laser in vivo. METHODS: The upper nasal limbus area next to the upper rectus muscle in right eyes received 10 minutes He-Ne laser irradiation (200mW/cm2) every day for three days, the left eyes served as control. Twenty-four hours after the last irradiation, both eyes of the rabbits were took trabeculectomy surgery. The expressions of CTGF in the filtration area were tested on the 7th, 14th and 28th day after surgery and collagen density was tested on the 14th and 28th day after surgery. Each of the time point had 7 rabbits. RESULTS: The expression of CTGF was lower than that of the control group's on the 7th and 14th day after trabeculectomy surgery (P=0.01, P=0.005). When examined on the 14th and 28th day, the collagen density of irradiation group were significantly lower than that of the control group's (P=0.013, P=0.01). CONCLUSION: Pretreating the filtration area with 200mW/cm2 He-Ne laser may be helpful in preventing scar formation after trabeculectomy in rabbit, possibly due to downregulation of the expression of CTGF and collagen synthesis in fibroblasts. He-Ne laser may be developed into a new scar preventing method in filtration surgery.     

The influence of He-Ne laser on scar formation after trabeculectomy in rabbits

AIM: To investigate the influence of He-Ne lasers on scar formation in the filtration canal after trabeculectomy in a rabbit model, as well as to explore the mechanisms for preventing scar formation when using He-Ne lasers in vivo. METHODS: Experiment 1: Four groups were established (four eyes in each group). In 12 eyes, the upper nasal limbus area next to the upper rectus muscle received 10 minutes of He-Ne laser irradiation (100, 150, 200mW/cm2; 60, 90, 120J/cm2) every day for three days. Four eyes served as controls. Twenty-four hours after the final irradiation, the rabbits were sacrificed and the irradiated tissue was excised, fixed with paraformaldehyde and tested for proliferating cell nuclear antigen (PCNA), connective tissue growth factor (CTGF) and apoptosis (TUNEL). Experiment 2: Forty-two rabbits were randomly divided into two groups and standard trabeculectomy was performed in the right eyes either after 200mW/cm2 He-Ne laser irradiation or not in the filtration area. The expression of PCNA and CTGF, apoptosis and collagen density in the filtration area were tested on the 7th, 14th and 28th day after surgery. RESULTS: Experiment 1: There were no more PCNA and CTGF positive cells in the He-Ne irradiation group than in the control group. No apoptotic cells were found in either group. Experiment 2: The expression of PCNA and CTGF was lower in the He-Ne irradiation group than in the control group on the 7th and 14th day after trabeculectomy surgery (P＜0.05); no apoptotic cells were detected in either group. Collagen density was significantly lower in the He-Ne irradiation group than in the control group on the 14th and 28th day after surgery (P＜0.05＝. CONCLUSION: Pretreating the filtration area with 200mW/cm2 (120J/cm2) of He-Ne laser irradiation may be helpful in preventing scar formation after trabeculectomy, possibly due to the downregulation of the expression of PCNA, CTGF and collagen synthesis in fibroblasts.     

紫杉醇在小梁切除术中的应用及其抗增生作用的研究

背景青光眼滤过手术是控制高眼压的主要方法,而滤过泡的纤维增生是手术失败的主要原因。寻找安全、有效的抗瘢痕药物是抗青光眼研究的主要目标之一。目的探讨紫杉醇在小粱切除术中的应用,评价紫杉醇对结膜下滤过泡的抗增生作用。方法32只家兔双眼行标准小梁切除术,随机分为生理盐水组、0．3g／L丝裂霉素C（MMC）组、0．2g／L紫杉醇组及0．3g／L紫杉醇组,每组8只兔16只眼。滤过手术术中巩膜瓣下分别用生理盐水、MMC、0．2g／L紫杉醇及0．3g／L紫杉醇浸泡3min。术后4、7、14、28d用Schlitz眼压计测量眼压、裂隙灯下观察滤过泡情况,并分别在上述各时间点处死2只兔,摘除动物眼球,取5mmx5mm滤过道处组织行苏木精一伊红染色、Massoni色染色比较各组滤过道开放情况、炎性细胞数量和新生胶原纤维的增生情况。结果手术前及手术后第4天,4个组兔眼压的总体比较差异均无统计学意义（F=0．54,P=0．83;F=0．57,P=0．87）。术后第7、14、28天,MMC组、0．2g／L紫杉醇组及0．3g／L紫杉醇组眼压明显低于生理盐水组,差异均有统计学意义（P〈0．05）;0．3g／L紫杉醇组眼压值均低于其他3组,差异均有统计学意义（P〈0．05）。术后第4天,4个组兔手术眼均为功能型滤过泡,术后第7、14、28天,MMC组、0．2g／L紫杉醇组功能型滤过泡的眼数均明显多于生理盐水组,但少于0．3g／L紫杉醇组,差异均有统计学意义（P〈0．05）。苏木精一伊红染色表明,术后生理盐水组滤过道炎性细胞数量明显多于其他3组,0．3g／L紫杉醇组炎性细胞浸润最少。Masson染色显示,0．3g/L紫杉醇组新生胶原纤维增生情况明显少于其他3组。结论小梁切除术中局部应用紫杉醇能抑制炎性细胞、胶原纤维的增生,有助于功能型滤过泡的形成,可有效降低眼压。     

重组人干扰素α-2b原位凝胶滴眼剂对兔小梁切除术后滤过瘢痕的影响

目的与传统干扰素α-2b(IFNα-2b)滴眼液相比,观察IFNα-2b原位凝胶滴眼剂在家兔实验性滤过手术后的抗瘢痕化作用。方法选取健康新西兰大白兔10只,随机平均分为两组。全部家兔均双眼进行标准小梁切除手术。右眼作为实验眼,左眼作为对照眼。手术第2天起右眼滴用IFNα-2b原位凝胶或滴眼液,4次/d,持续至术后14d。观察术前、术后眼压变化。术后28d,处死所有家兔,取滤过区组织,进行苏木素-伊红(HE)染色,观察滤过区瘢痕形成情况。结果除术后第2周滴用IFNα-2b滴眼液组双眼眼压差异无统计学意义(P=0.145)外,其余术后各时间点,两组家兔眼压右眼均低于左眼,差异有统计学意义(P<0.05)。术后各时间点,使用原位凝胶眼的眼压均低于使用滴眼液眼的眼压,但是差异并不具有统计学意义(P>0.05)。滤过区HE染色发现IFNα-2b滴眼液组巩膜瓣下纤维结缔组织明显较对照组少,成纤维细胞相对较少,并可见部分滤过泡间隙。IFNα-2b原位凝胶组可见巩膜瓣下结缔组织形态较规则,成纤维细胞较少,可见明显滤过泡间隙。所有手术眼均未发现明显眼部炎症表现。结论 IFNα-2b原位凝胶及IFNα-2b滴眼液均能显著抑制家兔实验性滤过性手术后的瘢痕形成。IFNα-2b原位凝胶的抗瘢痕作用略优于滴眼液。     

Control of filtering bleb structure through tissue bioengineering: An animal model

PURPOSE: To devise a means of providing controlled resistance between the anterior chamber and the subconjunctival space after trabeculectomy by implantation of a biodegradable, porous collagen matrix. METHODS: Matrices were implanted in the right eyes of 17 rabbits after trabeculectomy, while left eyes served as surgical controls. The scleral flap was sutured loosely, and the implant provided pressure on the scleral flap to reduce overfiltration. Trabeculectomy in the control eyes was performed with tight sutures using standard methodology. Intraocular pressure (IOP) was measured before surgery and on days 3, 7, 14, 21, and 28 after surgery. Masson trichrome and alpha-smooth muscle actin stains were used for histologic study of the filtering blebs. RESULTS: The initial postoperative IOP reduction was approximately equal, at 14% to 16%, for both groups. In the implanted group, the IOP continued to decrease to 55% below baseline at day 28 as the implant gradually degraded. In the control group, IOP had returned to the preoperative level by day 21. Histologic examination with Masson trichrome and alpha-smooth muscle actin stains showed a prominent bleb in the implanted group compared with scar formation and limited bleb formation in the control group. CONCLUSIONS: Implantation of a biodegradable, porous collagen matrix in the subconjunctival space offers the potential for a new means of avoiding early scar formation and maintaining long-term IOP control by creating a loosely structured filtering bleb.     

兔眼小梁切除术后结缔组织生长因子的表达

目的 研究结缔组织生长因子(CTGF)在兔眼小梁切除术后滤过泡中的表达及意义.方法 对照实验研究.将49只健康家兔分为5组:正常眼未手术组(A组),高眼压未手术组(B组),高眼压模型小梁切除术组(C组),正常眼小梁切除术组(D组),正常眼假手术组(E组),并分别在术后第2、5、7、14及21天,取右眼滤过泡部位的球结膜和浅层巩膜组织,A组和B组作为对照组.采用RT-PCR法检测CTGF mRNA的相对表达量,应用免疫组织化学方法 检测CTGF蛋白表达水平,HE染色行组织病理学检查.采用SAS 9.1.3统计学软件,对逆转录聚合酶链反应(RT-PCR)和免疫组织化学检测结果 采用析因设计定量资料方差分析.结果 C、D、E组术后CTGF表达水平高于A组和B组,术后第5天CTGF表达水平达到高峰,与其他时间点的表达差异有统计学意义(F=19.54,P<0.05);术后第2和5天,CTGF mRNA表达水平,C组高于同期D组(t=2.300,5.140;P<0.05);术后第2、5、7、14天,D组高于同期E组(t=-2.927,-6.424,-4.176,-4.997,P<0.05).CTGF蛋白表达水平,术后第2、5、7、14天,C组高于同期D组(t=-7.147,-10.955,-9.900,-6.385,P<0.05);术后第2、5、7、14天,D组高于同期E组(F=68.33,P<0.05).组织病理学检查结果 显示,术后第5天,C、D、E组局部炎性反应达高峰,表现为中性粒细胞、单核细胞及巨噬细胞浸润,成纤维细胞增殖达高峰.结论 小梁切除术后家兔CTGF表达水平的变化趋势表明,CTGF在伤口愈合及瘢痕形成过程中可能起重要作用,高眼压状态可促进CTGF表达水平上调.(手华眼科杂志,2009,45:168-174)     

应用滤过性手术在大鼠眼上建立结膜滤过泡瘢痕化模型的研究

目的探讨应用滤过性手术在大鼠眼上建立结膜滤过泡瘢痕化模型的可行性。方法选用35只健康成年SD大鼠作为实验动物,在全身麻醉下行单侧眼球滤过性手术,术后观察结膜滤过泡的形成情况,并观察眼前节反应;手术后1、3d,1、2、4周共5个时间点各处死3只大鼠,摘除其眼球,常规10％福马林固定,切片,HE染色,观察其病理改变,并以对侧眼作为正常对照。结果所有大鼠在手术后7d均有结膜滤过泡形成;手术后8d,结膜滤过泡消失,最长的结膜滤过泡能够维持21d。组织病理学观察显示,术后1d,结膜下组织疏松水肿,血管扩张、充血明显,有较多中性粒细胞浸润;术后3d,结膜下组织血管扩张、充血,除有中性粒细胞浸润外,还可见单核巨噬细胞浸润,结缔组织疏松、水肿较前减轻;术后7d,血管扩张充血程度较术后3d时减轻,有较多的纤维母细胞增生,纤维母细胞的细胞核呈椭圆形、胞质丰富,在纤维母细胞周围,可见少量疏松的胶原纤维形成;术后2周,纤维母细胞数量较术后1周时有所减少,细胞变细长,细胞核两端变尖,周围胶原纤维较术后1周时致密;术后4周时,结膜下组织血管多数闭合,细胞成分明显减少,纤维母细胞演变为纤维细胞,细胞核变细长,周围伴大量胶原纤维沉积,呈现瘢痕样改变。结论应用滤过性手术在大鼠眼上建立结膜滤过泡瘢痕化模型具有可行性,其病理改变是一个创伤修复的过程,该模型的建立不仅为青光眼手术后结膜滤过泡瘢痕化的防治,同时也为防治其他原因导致的结膜瘢痕化提供了新的研究方法。     

大蒜素对青光眼滤过术后兔眼滤过泡瘢痕化的抑制作用

目的　探讨大蒜素对青光眼滤过术后兔眼滤过泡瘢痕化的抑制作用。方法　取健康同品系大白兔36只72眼,随机分成大蒜素组、丝裂霉素组、生理盐水组,每组24眼,均行小梁切除术。各组分别将浸有大蒜素（1.112 kg·L^-1）、丝裂霉素（0.3 g·L^-1）、生理盐水的棉片放置于结膜囊-巩膜瓣-巩膜床层间,5 min后完全取出棉片,充分冲洗。术后各组分别测眼压,观察滤过泡的形态,计算滤过泡面积并行眼前节照相。术后28 d处死各组兔,行HE染色及Masson染色对比观察各组结膜下瘢痕增生情况。结果　术后第1天3组眼压较术前降低,随后眼压逐渐回升。各组不同时间内的眼压差异均有统计学意义（均为P<0.05）。术前及术后1 d、4 d、7 d、14 d两两比较中除术后4 d丝裂霉素组眼压比生理盐水组低（P=0.025）外,其余各时间眼压两两比较,差异均无统计学意义（均为P>0.05）。术后28 d生理盐水组眼压高于大蒜素组和丝裂霉素组（均为P<0.05）,大蒜素组和丝裂霉素组相比差异无统计学意义（P>0.05）。术后1~4 d 3组滤过泡面积分别达到最大值,之后滤过泡面积逐渐减小。统计学分析结果示3组滤过泡面积减小量在不同的时间点上是不同的（P<0.05）。不同分组之间的滤过泡面积减小量有明显差异（P<0.05）。术后大蒜素组与丝裂霉素组滤过泡面积减小幅度相同,差异无统计学意义（P>0.05）,生理盐水组比大蒜素组和丝裂霉素组的滤过泡面积减小速度更快,差异均有统计学意义（均为P<0.05）。术后28 d各组结膜上皮均完整无破损及增厚。HE染色显示大蒜素组和丝裂霉素组兔眼瘢痕组织较少,滤过通道较宽;生理盐水组兔眼瘢痕组织较多,滤过通道狭窄。Masson染色显示大蒜素组和丝裂霉素组可见少量炎性细胞浸润,滤过区成纤维细胞及胶原纤维疏松,结膜及巩膜间隙较宽;生理盐水组可见大量炎性细胞浸润,滤过区成纤维细胞及胶原纤维致密,结膜及巩膜间隙较窄,形成大量的瘢痕组织。术后 28 d 根据胶原纤维占整个结膜和巩膜空间总面积的百分比分析,大蒜素组［（58.13±5.33）%］比丝裂霉素组［（62.42±2.95）%］胶原纤维少,差异有统计学意义（P<0.05）,大蒜素组和丝裂霉素组均比生理盐水组［（89.75±3.91）%］胶原纤维少,差异均有统计学意义（均为P<0.05）。结论　大蒜素和丝裂霉素有着相同的功效,可通过抑制小梁切除术后滤过泡瘢痕化,对功能性滤过泡有一定的维持作用。     

Fibrin glue as a suture substitute: histological evaluation of trabeculectomy in rabbit eyes

PURPOSE: To describe the outcome of the use of fibrin adhesive (Quixil) in penetrating trabeculectomy in a rabbit model. METHODS: Fibrin adhesive was used experimentally to attach the conjunctiva and the scleral flap in two groups of 17 New Zealand albino adult rabbits (34 eyes). In the first experiment (20 eyes), the fibrin adhesive was used to reattach the tissue after conjunctival peritomy and scleral flap only in 14 eyes (experiment I). In 6 eyes (controls), the conjunctiva was attached with nylon sutures. In the second experiment (14 eyes), the fibrin adhesive was used after conjunctival peritomy, scleral flap, and penetrating trabeculectomy in 8 eyes (experiment II). In a control group of 6 eyes, nylon sutures were used to attach the scleral flap and the conjunctiva after penetrating trabeculectomy. Biomicroscopy and histopathological examinations were performed on postoperative days 1, 3, 7, 14, 21, and 30. Intraocular pressure was measured before and after surgery in the second experiment. Main outcome measures are histological presence of adhesive in the tissue, degree of capillary congestion, inflammatory reaction, collagen density [scar formation] and clinical (IOP measurements before and after surgery, conjunctival chemosis, anterior chamber reaction, presence of filtering bleb and wound leakage). RESULTS: In experiments I and II, the adhesive was well identified histologically in the tissue as an amorphic eosinophilic substance for up to day 3 and nearly disappeared by day 7. An acute inflammatory reaction was noted for up to 14 days, which converted to chronic inflammation with collagen deposits and scar formation by day 30. Similar inflammatory reaction was observed in the control group. The adhesive had no adverse effects on ocular tissue compared with sutures. One eye in experiment II demonstrated wound dehiscence. Intraocular pressure dropped from 17.35 mmHg preoperatively to 8.28 mmHg on postoperative day 1 in experiment II, and from 17.2 mmHg to 11.5 mmHg in the controls. No significant change in intraocular pressure was noted in experiment I. CONCLUSIONS: The fibrin adhesive had no adverse effects on ocular tissue compared with sutures. It might serve as an effective substitute for conjunctival and scleral wound closure in trabeculectomy surgery.