糖尿病大鼠视网膜神经节细胞凋亡及补肾活血中药抑制其凋亡的机制

The mechanism of apoptosis and effect on inhibition in retinal ganglion cells of rats with diabetes mellitus and its inhibition by traditional Chinese medicine for bushenhuoxue(补肾活血)therapy

目的 探讨糖尿病大鼠视网膜神经节细胞 (RGC)凋亡及补肾活血中药抑制凋亡的机制。方法 以链脲佐菌素造成大鼠糖尿病模型 ,给予补肾活血中药加降糖灵混悬液 ,连续 6月 ,采用甲苯胺蓝 -伊红染色及免疫组化染色结合图像分析半定量检测RGC数目、尼小体、bcl-2、bax、NT -3的含量 ,计算bcl-2 /bax蛋白的比值 ,并使用逐步引入剔除模型进行多元线性回归分析。结果 与正常组相比 ,模型组RGC数目明显减少 (P <0 0 0 1 ) ,尼氏小体、bcl-2、NT -3、bcl-2 /bax蛋白含量显著降低 (P <0 0 0 1 ) ,bax含量显著增高 (P <0 0 0 1 ) ;与模型组相比 ,补肾活血中药组RGC细胞数目明显增多 (P <0 0 0 1 ) ,尼小体、bcl-2、NT -3、bcl-2 /bax蛋白含量显著增高 (P <0 0 0 1 ) ,bax含量显著降低 (P <0 0 0 1 )。回归分析结果显示RGC数目和尼小体含量分别与bcl-2 /bax蛋白比值呈正相关 ,前者r=0 754,P <0 0 0 1 ,后者r=0 578,P<0 0 0 1。结论 糖尿病大鼠RGC凋亡及补肾活血中药抑制其凋亡的可能机制之一是通过调节bcl-2 /bax的比值 ,神经营养因子NT -3量的改变也有一定的作用。

OBJECTIVE To investigate the mechanism of apoptosis of retinal ganglion cell (RGC) on the diabetic rats and it was inhibited by traditional Chinese medicine for ushenhuoxue(补肾活血)therapy.METHODS The animal model of diabetes mellitus was introduced by streptozotocin, then gave suspension of the traditional Chinese medicine for bushenhuoxue therapy and phenformin for six months. The number of RGC and the content of the Nissl body and B cell lymphoma, leukemia 2 (bcl-2), and Bcl-2 associated X protein (bax) and Neurotrophin-3 (NT-3) protein were measured semi-quantitatively with toluidine blue-eosin staining and immunohistochemistry methods combined with image analysis system. Ratio of bcl-2/bax protein was counted, then linear multianalysis regression was used for stepwise method. RESULTS Compared with normal group, the number of RGC reduced obviously in the model group(P<0.001), and the content of the Nissl body and the bcl-2 and NT-3 and bcl-2/bax protein decreased notably (P<0.001), and the content of the bax protein increased notably in the model group(P<0.001).Compared with model group, the number of RGC increased obviously (P<0.001), and the content of the Nissl body and the bcl-2 and NT-3 and bcl-2/bax protein increased notably (P<0.001), and the content of the bax protein decreased notably in the bushenhuoxue group (P<0.001). The result of regression analysis showed that the number of RGC and the content of the Nissl body was a positive relation to ratio of bcl-2/bax protein, the former :r=0.754,P<0.001; and the latter r=0.578 P<0.001. CONCLUSION The mechanism of apoptosis of retinal ganglion cell in diabetic rats and that was inhibited by traditional Chinese medicine for bushenhuoxue therapy was mainly of adjusting ratio of bcl-2/bax protein, then changes of content of neurotrophic factor NT-3 bringed partial effects into play.