摘要
目的:
1.建立大鼠白内障模型,并探讨亚硒酸钠诱导大鼠白内障模型晶状体混浊的发生机制。
2.探讨祛障穴冷冻疗法和改变冷冻位点对亚硒酸钠诱导大鼠白内障模型晶状体混浊进展的抑制和延缓机制。
3.通过对祛障穴冷冻疗法和改变冷冻位点方式的对比分析,探索优势冷冻位点,并探讨后者针对年龄相关性白内障治疗的有效性和可行性。
4.为低温生物医学揭示冷冻对白内障抑制延缓作用的机理提供宝贵资料。
方法:
对35只大鼠行裂隙灯检查,排除晶状体病变。随机抽取5只大鼠作为正常对照组,其余大鼠随机分为3组,每组10只,隔日于颈背部皮下注射1mmol/ L的亚硒酸钠,共注射3次,正常组给予等剂量0.9%生理盐水,监测大鼠晶状体混浊变化。待晶状体刚刚出现混浊时,祛障穴冷冻治疗组大鼠给予祛障穴冷冻疗法,每周1次,5周一疗程,方法参照李永才发明祛障穴冷冻疗法。冷冻位点改变治疗组大鼠,于大鼠角巩膜缘12点位每次随机抽取4位点,行冷冻治疗,操作方法同祛障穴冷冻治疗组。未治疗组不予处理。通过裂隙灯观察并记录大鼠晶状体混浊的进展。晶状体混浊分级标准采用英国牛津大学眼科实验室方法。6wk时处死大鼠,取出晶状体,测定其中水溶性蛋白质含量、谷胱甘肽(GSH)含量、谷胱甘肽还原酶(GR)及过氧化氢酶(CAT)活性。
结果:
1.除正常组以外,其余三组大鼠双眼均于1wk开始出现混浊,2wk~6wk白内障未治疗组大鼠双眼晶状体混浊进展较快,而两治疗组大鼠双眼晶状体相对B组混浊进展较慢,且混浊程度轻于B组,具有高度统计学意义(右:P=0.000,左:P=0.002)。
2.晶状体可溶性蛋白质含量,与正常组相比,未治疗组大鼠显著降低,有高度统计学意义(右:P=0.010,左:P=0.001),祛障穴冷冻组右眼、冷冻位点改变组双眼低于正常组,但无统计学意义(P>0.05),祛障穴冷冻组左眼眼较正常组显著降低,具有高度统计学意义(P=0.016)。
3.GR活性,各组大鼠双眼晶状体比较其差别无统计学意义(P>0.05);
4.GSH比较,与正常组大鼠双眼比较白内障未治疗组、祛障穴冷冻组和冷冻位点改变治疗组双眼晶状体GSH含量明显降低,有高度统计学意义(P=0.000,P=0.000,P=0.000)。
5.CAT活性,与正常组大鼠双眼比较白内障未治疗组、祛障穴冷冻组和冷冻位点改变治疗组双眼晶状体CAT活性明显降低,有高度统计学意义(P=0.000,P=0.000,P=0.000)。祛障穴冷冻组和冷冻位点改变治疗组CAT活性高于白内障未治疗组,右眼分别为41.4%和35.9%(P=0.003,P=0.012),左眼分别为32.3%和27.4%(P=0.008,P=0.031),具有高度统计学意义。
6.色素沉着,祛障穴冷冻组和冷冻位点改变治疗组在对大鼠角巩膜缘进行冷冻治疗瞬间均形成冻斑,随即冻斑消失,6wk少数祛障穴冷冻组大鼠角巩膜缘冷冻位点处出现不同程度色素沉着,而这一表现在冷冻位点改变治疗组大鼠中未能出现。
结论:
1.亚硒酸钠诱导大鼠白内障模型是白内障发病机制及疗效评价的可靠模型。
2.晶状体内氧化损伤可能贯穿于白内障的发生发展。
3.祛障穴冷冻疗法对大鼠初发期白内障晶状体混浊有较好的抑制、延缓作用
4.祛障穴冷冻疗法与随机改变4冷冻位点在对白内障大鼠晶状体混浊的抑制、延缓作用的比较中无显著性差异,而后者能够更好的分散由冷冻对局部组织引起的刺激和伤害。
5.依据祛障穴冷冻疗法治疗老年性未成熟期白内障规范化整理研究成果(国家中医药管理局诊疗技术课题,国中医药科2001ZL27号),预计角巩膜缘随机选取4冷冻位点冷冻治疗年龄相关性白内障将取得较好疗效。
Purpose:
1. Cataract model in rats, and sodium selenite cataract model induced by the mechanism of lens opacity.
2. To explore Qu Zhang Point cryosurgery and changing the freezing point sites on the selenite cataract model induced by inhibition of the progress of lens opacity and delay mechanisms.
3. By Qu Zhang Point cryosurgery and changing the freezing point cryotherapy sites the way comparative analysis of the latter effect for the treatment of senile cataract the predictability and feasibility.
Methods:
35 rats were slit lamp examination to exclude the lens disease. 5 rats were randomly selected as normal control group, the remaining rats were randomly divided into 3 groups of 10 each, every other day subcutaneous injection in the neck, 1mmol / L of sodium selenite were injected three times the normal group was given 0.9% saline, monitoring changes in lens opacity. Lens opacity appears to be just when the treatment group of Qu Zhang Point cryosurgery to cured by Qu Zhang Point cryosurgery, 1 time a week, 5 Mon treatments, methods, invented by Mr Li You cai’Qu Zhang Point cryosurgery. Freezing changes the treatment sites of the rats in the rat limbal 12 randomly selected 4-bit each time point to cryosurgery, use the same methods as Qu Zhang Point cryosurgery group. Through the slit lamp to observe and record the progress of rat lens opacification. Lens opacity classification criterion used by the laboratory method of Ophthalmology, University of Oxford. 6wk rats were sacrificed, remove the lens to determine which water-soluble protein, glutathione (GSH), glutathione reductase (GR) and catalase (CAT) activity.
Results:
1. In addition to the normal group than the other three rats eyes were cloudy began in 1wk, 2wk~6wk untreated cataract lens opacity group eyes rapid progress, while the two treatment group eyes the progress of lens opacities compared with the relative B group slow, and the turbidity and light in the B group, the difference was significant (right:P=0.000,.left:P=0.002).
2. Lens soluble protein content, compared with the normal group, no treatment group were significantly lower, with statistical significance (right: P = 0.010, left: P = 0.001), Qu Zhang Point cryosurgery right eye point freezing group, frozen sites change the group with normal control eyes, but no statistical significance (P>0.05), the left eye of Qu Zhang Point cryosurgery group were significantly lower than normal (P = 0.016).
3.GR activity, each rat eye lens compare the difference was not statistically significant (P>0.05);
4.GSH comparison, compared with normal rats of cataract eyes without treatment, Qu Zhang Point cryosurgery group and removing of point freezing treatment group,eyes sites to change the lens GSH concentration decreased and there was significant difference (P = 0.000, P = 0.000, P = 0.000).
5.CAT activity, compared with normal rats of cataract eyes without treatment, Qu Zhang Point cryosurgery group and removing of point freezing treatment group sites change CAT activity was significantly lower lens eyes, there was significant difference (P = 0.000, P = 0.000, P = 0.000). Qu Zhang Point cryosurgery group and removing of point freezing treatment group sites change CAT activity in the treatment group than in untreated patients with cataract, right eye were 41.4% and 35.9% (P = 0.003, P = 0.012), left eye were 32.3% and 27.4% ( P = 0.008, P = 0.031), with statistical significance.
6. Pigmentation, Qu Zhang Point cryosurgery group and removing of point freezing treatment group change in position on the rats in the treatment group were frozen treat limbal formation of cold spots are an instant, then cold spots disappeared, 6wk few Qu Zhang Point cryosurgery group limbal frozen rats frozen sites pigmentation appears at different levels, and this reflected in the frozen site failed to change the treatment group there.
Conclusion:
1. Selenite cataract model induced cataract pathogenesis and treatment evaluation of reliableModel.
2. The lens oxidative damage may run through the development of cataracts.
3. Qu Zhang Point cryosurgery group onset of cataract in rat lens opacity has good inhibition, delay Slow effect.
4. Qu Zhang Point cryosurgery group and removing of point freezing of randomly selecting for each 4 sites treatment group Limbal rats were in dialogue, impaired inhibition of rat lens opacity and delay comparison of the role of no significant difference, while the later can be better distributed by the freezing of the local tissue irritation and damage caused.
5. According to dispel the Qu Zhang Point cryosurgery treatment of senile cataract is not mature research results standardized order (State Administration of Traditional treatment technology issues, the National Bureau 2001ZL27 number of Chinese medicine), is expected to limbal frozen each 4 sites were randomly selected treatment age-related cataract will get better effect.
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