弱激光疗法对兔感染性眼内炎抗炎作用研究
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摘要
感染性眼内炎(infectious ophthalmitis)是一种眼内感染,是眼球穿孔伤和内眼手术后最严重的并发症。该疾病进展快,破坏性强。病原微生物本身及其释放的毒素引起的强烈炎症反应一直是眼内炎治疗面临的困境。强烈的炎症反应导致眼内血管通透性增强,中性粒细胞、巨噬细胞迁移以及蛋白的渗漏,在很大程度上引起视网膜结构破坏性改变,导致不可逆损伤,影响视功能。在抗菌素使用的前提下,有效地控制眼内炎症反应也是治疗的关键所在。治疗时机、治疗方法的选择与预后紧密相关。糖皮质激素被广泛应用于感染性眼内炎治疗。但是,炎症反应是机体的一种防御功能,因此糖皮质激素在抑制炎症、减轻症状的同时,也降低了机体的防御,可导致感染扩散。近几年研究发现弱激光疗法(low-level laser therapy, LLLT)在各种急性非感染性炎症过程中,可减轻炎症的渗出、水肿、毛细血管扩张、白细胞浸润和吞噬等反应,从而改善炎症状态。本研究通过建立感染性眼内炎模型,探索LLLT在感染性眼内炎治疗过程中的安全性、有效性,并对其可能的作用机制进行初步研究。
目的：
本课题利用标准表皮葡萄球菌菌株建立兔眼感染性眼内炎模型,在玻璃体腔注射敏感抗菌素干预条件下,采用LLLT、糖皮质激素(地塞米松)抗炎治疗。这一研究试图探索感染性眼内炎LLLT治疗的安全性、有效性,为LLLT治疗该疾病提供实验依据,以期减少、替代糖皮质激素在感染性眼内炎抗炎治疗过程中的应用。
方法：
1.不同剂量弱激光经瞳孔照射兔眼,通过眼科常规检查及电生理检查,探讨弱激光疗法的安全性。
2.采用灭活的金黄色葡萄球菌混悬液建立兔眼内炎模型,采用弱激光疗法进行治疗,探讨弱激光疗法的有效性。
3.建立表皮葡萄球菌性兔眼内炎模型,在万古霉素干预下,比较地塞米松、LLLT在细菌性眼内炎治疗过程中抗炎作用。
4.建立烟曲霉兔眼内炎模型,在两性霉素B干预下,比较地塞米松、LLLT在真菌性眼内炎治疗过程中的抗炎作用。
5.对PMA刺激的人白血病细胞系HL-60行LLLT,通过ELISA方法测定TNF-α,对LLLT可能的抗炎机制初步探讨。
结果：
1.10mw×10min组ESR未见明显视网膜功能不可逆损伤,适宜剂量LLLT用于眼内照射是安全的。
2.对细菌毒素导致的兔眼内炎,LLLT可明显减低炎症反应,对视网膜功能具有保护作用(P<0.05)。
3.表皮葡萄球菌性眼内炎,在万古霉素干预下,不同时间点地塞米松、LLLT均表现出明显的抑制炎症反应作用,可减轻炎症反应(P<0.05)。
4.成功建立烟曲霉兔眼内炎模型,但是由于烟曲霉性眼内炎炎症反应剧烈,两性霉素治疗干预后内眼反应更为剧烈,短时间内即发展为全眼球炎,故无法评估地塞米松、LLLT的抗炎作用。
5.经LLLT处理的PMA刺激的人白血病细胞系HL-60产TNF-α减低(P>0.05)。LLLT抗炎作用机制可能是抑制细胞因子TNF-α生成,细胞因子间相互影响有待进一步研究。
结论：
本论文的研究显示不论在细菌毒素诱导的眼内炎还是表皮葡萄球菌性眼内炎治疗过程中,LLLT显示出良好的抗炎效果,可以应用于眼内炎抗炎治疗。
Infectious endophthalmitis is the most serious complication after intraocular surgery and penetrating eye injuries. Infectious endophthalmitis can quickly damage eye tissue and visual function, and results in loss of vision and ocular atrophy if the inflammation is not effectively controlled. It is now generally believed that tissue damage and intraocular inflammation are closely related. Therefore, anti-inflammatory treatment is as important as anti-microbial treatment to control disease progression and protect visual function. Dexamethasone is widely used to treat endophthalmitis. However, some researchers believe that dexamethasone can damage the immune system, and may accelerate intraocular infection and the destruction of intraocular tissue.
In recent years, low-level laser therapy (LLLT) has been widely applied in medicine. Many studies have found that LLLT has a significant biological regulatory role in the inflammatory process, such as the ability to reduce the speed and extent of inflammatory exudation, improved capillary permeability to promote absorption and dissipation of inflammation, etc.
Objective
In this study, a rabbit endophthalmitis model for Staphylococcus epidermidis infection was established, and either LLLT or dexamethasone was administered in combination with vancomycin to determine the anti-inflammatory effect of LLLT on infectious endophthalmitis and to assess the feasibility of LLLT to reduce or replace dexamethasone in the treatment of infectious endophthalmitis.
Methods
1. Different dose laser exposured to rabbit's retina through pupil, then routine ophthalmitic examination and F-ERG were practiced in left rabbit eyes to investigate the safty of LLLT.
2. To inject inactived staphylococcu aureus into rabbits' left eyes and establish endophthamitis model. LLLT was used to relieve inflammation. To investigate the effectivity of LLLT in infectious endophthalmitis (P<0.05).
3. To inject staphylococcu epidermidis into rabbits'left eyes and establish endophthamitis model. With the injection of Vancomycin into vitreous dexamethasone and LLLT was usded to relieve inflammation (P<0.05).
4. To inject Aspergillus fumigatus into rabbits'left eyes and establish endophthamitis model. With the injection of Amphotericin B into vitreous cavity to antisepticize, dexamethasone and LLLT was usded to relieve inflammation.
5. LLLT expoure to HL-60 provocatived by PMA, the concertration of TNF in the supernatant solution was determined by ELISA.
Results:
1. Suitable dose LLLT is safe. The results of F-ERG didn't show the inreversible damage of retina (P<0.05).
2. LLLT relieved the inflammmation obviosly of the endophthlmitis induced by bacteriotoxin and protected the function of retina (P<0.05).
3. With the effect of vancomycin, LLLT and dexamethasone showed the obvious anti-inflammation role to Staphylococcus epidermidis endophthalmitis at different time (P<0.05).
4. Established the Aspergillus fumigatus endophthalmitis successfully, the inflammation of Aspergillus fumigatus endophthalmitis was very strong and developed panophthalmitis in a short time, so it was difficult to investigate the anti-inflammation of LLLT to Aspergillus fumigatus endophthalmitis.
5. The concertration of TNF-αproduced by HL-60 expoured to LLLT was lower (P>0.05).
Conclusion:
LLLT showed a good anti-inflammation in endophthalmitis induced by inactived staphylococcu aureus or staphylococcu epidermidis. LLLT can be used to treat infectios endophthalmitis.
目的：
本课题利用标准表皮葡萄球菌菌株建立兔眼感染性眼内炎模型,在玻璃体腔注射敏感抗菌素干预条件下,采用LLLT、糖皮质激素(地塞米松)抗炎治疗。这一研究试图探索感染性眼内炎LLLT治疗的安全性、有效性,为LLLT治疗该疾病提供实验依据,以期减少、替代糖皮质激素在感染性眼内炎抗炎治疗过程中的应用。
方法：
1.不同剂量弱激光经瞳孔照射兔眼,通过眼科常规检查及电生理检查,探讨弱激光疗法的安全性。
2.采用灭活的金黄色葡萄球菌混悬液建立兔眼内炎模型,采用弱激光疗法进行治疗,探讨弱激光疗法的有效性。
3.建立表皮葡萄球菌性兔眼内炎模型,在万古霉素干预下,比较地塞米松、LLLT在细菌性眼内炎治疗过程中抗炎作用。
4.建立烟曲霉兔眼内炎模型,在两性霉素B干预下,比较地塞米松、LLLT在真菌性眼内炎治疗过程中的抗炎作用。
5.对PMA刺激的人白血病细胞系HL-60行LLLT,通过ELISA方法测定TNF-α,对LLLT可能的抗炎机制初步探讨。
结果：
1.10mw×10min组ESR未见明显视网膜功能不可逆损伤,适宜剂量LLLT用于眼内照射是安全的。
2.对细菌毒素导致的兔眼内炎,LLLT可明显减低炎症反应,对视网膜功能具有保护作用(P<0.05)。
3.表皮葡萄球菌性眼内炎,在万古霉素干预下,不同时间点地塞米松、LLLT均表现出明显的抑制炎症反应作用,可减轻炎症反应(P<0.05)。
4.成功建立烟曲霉兔眼内炎模型,但是由于烟曲霉性眼内炎炎症反应剧烈,两性霉素治疗干预后内眼反应更为剧烈,短时间内即发展为全眼球炎,故无法评估地塞米松、LLLT的抗炎作用。
5.经LLLT处理的PMA刺激的人白血病细胞系HL-60产TNF-α减低(P>0.05)。LLLT抗炎作用机制可能是抑制细胞因子TNF-α生成,细胞因子间相互影响有待进一步研究。
结论：
本论文的研究显示不论在细菌毒素诱导的眼内炎还是表皮葡萄球菌性眼内炎治疗过程中,LLLT显示出良好的抗炎效果,可以应用于眼内炎抗炎治疗。
Infectious endophthalmitis is the most serious complication after intraocular surgery and penetrating eye injuries. Infectious endophthalmitis can quickly damage eye tissue and visual function, and results in loss of vision and ocular atrophy if the inflammation is not effectively controlled. It is now generally believed that tissue damage and intraocular inflammation are closely related. Therefore, anti-inflammatory treatment is as important as anti-microbial treatment to control disease progression and protect visual function. Dexamethasone is widely used to treat endophthalmitis. However, some researchers believe that dexamethasone can damage the immune system, and may accelerate intraocular infection and the destruction of intraocular tissue.
In recent years, low-level laser therapy (LLLT) has been widely applied in medicine. Many studies have found that LLLT has a significant biological regulatory role in the inflammatory process, such as the ability to reduce the speed and extent of inflammatory exudation, improved capillary permeability to promote absorption and dissipation of inflammation, etc.
Objective
In this study, a rabbit endophthalmitis model for Staphylococcus epidermidis infection was established, and either LLLT or dexamethasone was administered in combination with vancomycin to determine the anti-inflammatory effect of LLLT on infectious endophthalmitis and to assess the feasibility of LLLT to reduce or replace dexamethasone in the treatment of infectious endophthalmitis.
Methods
1. Different dose laser exposured to rabbit's retina through pupil, then routine ophthalmitic examination and F-ERG were practiced in left rabbit eyes to investigate the safty of LLLT.
2. To inject inactived staphylococcu aureus into rabbits' left eyes and establish endophthamitis model. LLLT was used to relieve inflammation. To investigate the effectivity of LLLT in infectious endophthalmitis (P<0.05).
3. To inject staphylococcu epidermidis into rabbits'left eyes and establish endophthamitis model. With the injection of Vancomycin into vitreous dexamethasone and LLLT was usded to relieve inflammation (P<0.05).
4. To inject Aspergillus fumigatus into rabbits'left eyes and establish endophthamitis model. With the injection of Amphotericin B into vitreous cavity to antisepticize, dexamethasone and LLLT was usded to relieve inflammation.
5. LLLT expoure to HL-60 provocatived by PMA, the concertration of TNF in the supernatant solution was determined by ELISA.
Results:
1. Suitable dose LLLT is safe. The results of F-ERG didn't show the inreversible damage of retina (P<0.05).
2. LLLT relieved the inflammmation obviosly of the endophthlmitis induced by bacteriotoxin and protected the function of retina (P<0.05).
3. With the effect of vancomycin, LLLT and dexamethasone showed the obvious anti-inflammation role to Staphylococcus epidermidis endophthalmitis at different time (P<0.05).
4. Established the Aspergillus fumigatus endophthalmitis successfully, the inflammation of Aspergillus fumigatus endophthalmitis was very strong and developed panophthalmitis in a short time, so it was difficult to investigate the anti-inflammation of LLLT to Aspergillus fumigatus endophthalmitis.
5. The concertration of TNF-αproduced by HL-60 expoured to LLLT was lower (P>0.05).
Conclusion:
LLLT showed a good anti-inflammation in endophthalmitis induced by inactived staphylococcu aureus or staphylococcu epidermidis. LLLT can be used to treat infectios endophthalmitis.
引文
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