Abstract: Objective To explore the protective effect and mechanism of Bushenhuoxue(invigorating the kidney and promoting blood circulation) TCM serum on retinal ganglion cells (RGCs) purified cultured under the condition of hypoxia in vitro. Methods 30 neonatal rats 1d after birth were selected. Through the two step screening system, the RGCs of SD neonatal rats were purified cultured. 6 SD rats 7d after birth were selected and through improved enzymatic digestion technique, retinal Muller cells of neonatal SD rats were purified cultured. Then, according to the ratio of 1:25, RGCs were inoculated in the medium coated by retinal Muller cells to establish a common culture model. Glucose (50 mmol·L-1) and sodium dithionite (1.0 mmol·L-1) (high glucose hypoxia group), only glucose (50 mmol·L-1) (high glucose group) and sodium dithionite (1.0 mmol·L-1) (hypoxia group) were respectively added into the medium in vitro. Based on this, formulated Bushenhuoxue TCM serum of 20% volume fraction was respectively added for intervention. And the leakage of lactate dehydrogenase (LDH) and activity of glutamine synthetase (GS) in extracellular fluid of the model were respectively determined in 24 h, 48 h and 72 h. Results In 48 h, 72 h, the leakages of LDH in the normal control group, high glucose group and hypoxia group significantly increased, compared with that in 24 h (P <0.05). Comparing normal control group, high glucose group, hypoxia group and high glucose hypoxia group with their respectively corresponding TCM intervention in the same time, it could be found that the leakages of LDH were significantly higher than those TCM intervention groups (P <0.05). When it was 48 h and 72 h, the activity of GS in normal control group, high glucose group, hypoxia group, high glucose hypoxia group and their corresponding TCM intervention group increased significantly, compared with that in 24 h (P <0.05). With extension of time, the activity of GS in normal control group, high glucose group, hypoxia group, high glucose hypoxia group was significantly lower than that of the TCM intervention group. Conclusion High glucose or anoxic conditions can cause deviation of cell membrane stability of RGCs and retinal Muller cells and increase of permeability. Bushenhuoxue TCM can improve the stability of the cell membrane and increase GS activity, so as to protect the RGCs cells.

Key words: Bushenhuoxue TCM, hypoxia, retinal ganglion cells, retinal Muller cells