右归丸对糖尿病视网膜病变大鼠PI3K/Akt信号通路的影响

目的：探讨“阴中求阳”立法之右归丸对糖尿病视网膜病变大鼠PI3 K/Akt信号通路的影响。方法：SD大鼠80只,随机分组,10只为正常组,余大鼠通过一次性腹腔注射链脲佐菌素溶液(60 mg/kg )联合大鼠玻璃体腔注射VEGF(0.05μg)的方式建立糖尿病视网膜病变增殖期大鼠模型。模型建立成功后,将最终成模大鼠随机分为4组：右归丸高中低剂量组、模型组,右归丸高中低剂量组每天给予相应右归丸浓缩液浓度进行灌胃,模型组、正常组每天给予等剂量蒸馏水灌胃。连续灌胃3 mo后,采取SP免疫组织化学、Western-blot法分别观察检测大鼠视网膜组织PI3 K和Akt的表达情况。结果：免疫组织化学观察显示PI3 K和Akt免疫组化染色的阳性表达,在视网膜上均为棕黄色颗粒；正常组PI3 K和Akt表达部分分布于神经节细胞层,内核也有少量表达,呈弱阳性免疫反应；模型组、右归丸各治疗组大鼠视网膜神经节细胞层、内丛状层及内外颗粒层都有阳性表达；与模型组比较,右归丸中、高剂量组PI3 K和Akt表达均减弱,右归丸低剂量组表达减弱不明显；与右归丸低剂量组比较,右归丸中、高剂量组PI3 K和Akt表达减弱；右归丸中、高剂量组表达强弱比较无差异。 Western-blot检测结果显示与正常组比较,模型组、治疗组表达差异有显著统计学意义( P<0.01),模型组、治疗组PI3 K和Akt蛋白表达水平显著升高；与模型组比较,右归丸中、高剂量组表达差异有统计学意义(P<0.05),右归丸中、高剂量组PI3 K和Akt蛋白表达水平均降低,右归丸低剂量组表达差异无统计学意义(P>0.05)；与右归丸低剂量组比较,右归丸中、高剂量组表达差异有统计学意义(P<0.05),右归丸中、高剂量组PI3 K和Akt表达水平下降；右归丸中、高剂量组表达比较,差异无统计学意义(P>0.05)。结论：基于“阴中求阳”立法之右归丸可以通过影响PI3 K和Akt蛋白表达水平,抑制PI3 K/Akt 信号通路活化,延缓糖尿病视网膜病变的病变进程,为糖尿病视网膜病变防盲治疗提出新的治疗思路与科学依据。

Effects of a traditional Chinese patent medicine for PI3K/Akt pro-survival signal channel in diabetic retinopathy rat

?AIM: To investigate the effects of reinforcing yang from yin of a traditional Chinese patent medicine called YouguiWan for PI3K/Akt pro-survival signal channel in diabetic retinopathy rat.?METHODS:A total of 80 SPF SD rats were selected, 10 rats were selected to set up normal group. The remaining 70 rats were given streptozotocin (60mg/kg) peritoneal injection combined with intravitreal injection of VEGF ( 0. 05μg ) to establish proliferative diabetic retinopathy rat model. After the model successful, the rats were divided into four groups: YouguiWan high dose group, YouguiWan middle dose group, YouguiWan low dose group and model group. The treatment groups were fed daily YouguiWan concentrate, model group and the normal group received daily doses of distilled water.
After intragastric administration for 3mo, the PI3K, Akt expression of retinal tissue were observed by SP -immunohistochemistry and Western - blot method respectively.?RESULTS: Immunohistochemical observation showed PI3K, Akt expression immunohistochemical staining, were brown particles on the retina. Normal PI3K, Akt expression weakly positive immunological reaction unit located in the ganglion cell layer. The kernel was also a small amount of expression. Model group, each YouguiWan treatment group rat retinal ganglion cell layer, inner plexiform layer and the inner and outer layer of the particles had a positive expression. Compared with the model group, YouguiWan high dose group PI3K, Akt expression were decreased. YouguiWan low dose group decreased expression was not obvious. Compared with the low dose group, YouguiWan high dose group PI3K, Akt expression was decreased. The expression of the strength of the YouguiWan, high dose group showed no significant difference. Western-blot test results showed that compared with the normal group, model group, treatment group, the expression difference was statistically significant (P<0. 01). Model group, treatment group, PI3K, Akt protein levels were significantly increased. Compared with the model group, the YouguiWan high - dose group was statistically significant (P<0. 05). YouguiWan high dose group PI3K, Akt protein expression were decreased, YouguiWan low dose group was no significant difference ( P> 0. 05 ). Compared with the low dose group, YouguiWan high-dose group was statistically significant ( P < 0. 05 ). YouguiWan high dose group PI3K, Akt expression levels were decreased. YouguiWan high dose group was not statistically significant difference (P>0. 05).? CONCLUSION: Reinforcing yang from yin of YouguiWan can affect PI3K, Akt protein expression and inhibit PI3K/Akt signaling pathway, delay the disease process of diabetic retinopathy, propose new ideas and scientific basis for the treatment of diabetic retinopathy and blindness prevention treatment.