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目的:通过测定经臭氧灌肠治疗后的糖尿病大鼠视网膜及血清中血管内皮生长因子(VEGF)、缺氧诱导因子-1α(HIF-1α)与色素上皮衍生因子(PEDF)的表达差异,了解臭氧在早期糖尿病视网膜病变治疗的作用。方法:选取70只雄性SD大鼠中的10只做为正常对照组,予以正常饮食;余大鼠经适应性喂养后通过腹腔一次性注射链脲佐菌素(50mg/ml)制作糖尿病模型。将造模成功的大鼠模型随机分成模型对照组(B组)、氧气治疗组(C组)及臭氧治疗组(D组),其中臭氧治疗组给予臭氧灌肠治疗(2次/wk,共计1mo),氧气治疗组给予同等剂量及频次的氧气进行灌肠治疗,治疗1mo后取视网膜及血清,使用免疫组化法检测视网膜中VEGF的表达,使用ELISA及RT-PCR的方法检测视网膜及血清中VEGF、HIF-1α与PEDF的含量。结果:免疫组化结果显示VEGF主要在内层视网膜表达,而臭氧治疗组VEGF表达接近于空白对照组;血清与视网膜中VEGFmRNA表达在四组间均有统计学差异(F=23.923;P=0.000),其中臭氧治疗组VEGF的表达接近于空白对照组,但差异仍有统计学意义(P<0.05);而模型对照组同氧气治疗组的VEGF表达亦无统计学差异(P>0.05);相对于空白对照组,臭氧治疗组HIF-1α表达下降(P<0.05),而在模型对照组与氧气治疗组间无统计学差异(P>0.05)。PEDFmRNA在四组间的表达均有差异,但差异均无统计学意义(P>0.05)。结论:臭氧灌肠治疗可以有效的降低血清及视网膜中VEGF和HIF-1α的表达,可以一定程度的抑制早期糖尿病视网膜病变的发展。
OBJECTIVE: To determine the expression of vascular endothelial growth factor (VEGF), hypoxia inducible factor-1α (HIF-1α) and pigment epithelium-derived factor (PEDF) in the retina and serum of diabetic rats after ozone enema treatment Role in the treatment of early diabetic retinopathy. Methods: Ten of 70 male Sprague-Dawley rats were selected as the normal control group and were given normal diet. The rats were fed with streptozotocin (50mg / ml) via intraperitoneal injection after adaptive feeding. The successful model rats were randomly divided into model control group (group B), oxygen treatment group (group C) and ozone treatment group (group D). The ozone treatment group was treated with ozone enema (2 times / wk for a total of 1mo ). Oxygen treatment group was given the same dose and frequency of oxygen for enema treatment. Retinal and serum were taken after treatment for 1 month. The expression of VEGF in the retina was detected by immunohistochemistry. The expression of VEGF in retina and serum was detected by ELISA and RT-PCR , HIF-1α and PEDF content. Results: The results of immunohistochemistry showed that VEGF was mainly expressed in the inner retina, while the expression of VEGF in the ozone-treated group was close to that of the blank control group. The expression of VEGF mRNA in the serum and the retina was statistically different among the four groups (F = 23.923; P = 0.000 ). The expression of VEGF in the ozone treatment group was similar to that in the blank control group, but the difference was still statistically significant (P <0.05). However, there was no significant difference in VEGF expression between the model control group and oxygen therapy group (P> 0.05). Compared with the blank control group, the expression of HIF-1α in the ozone-treated group decreased (P <0.05), while there was no significant difference between the model control group and the oxygen-treated group (P> 0.05). PEDF mRNA expression in the four groups were different, but the difference was not statistically significant (P> 0.05). Conclusion: Oral enema treatment can effectively reduce the expression of VEGF and HIF-1α in serum and retina, which can inhibit the development of early diabetic retinopathy to a certain extent.