[Animal modeling] - Effect of membrane cells on granulosa cells injured by chemotherapy and intervention of Zuogui Pill containing serum

  Objective To investigate the effect of membrane cells on granulosa cells injured by chemotherapy and the intervention of Zuogui Pill containing serum.

  Methods The drug containing serum of Zuogui Pill was prepared, and the granulosa cells and membrane cells of rat ovary were cultured. The rats were divided into 8 groups: (1) blank control group; (2) Co culture group; (3) ZGW serum group; (4) ZGW serum+co culture group; (5) Model group; (6) Model+co culture group; (7) Model+ZGW serum group; (8) Model+ZGW serum+co culture group. The expression of Bax, Caspase-3, Beclin-1, p62, LC3B at the transcription level and translation level were detected by real-time fluorescent quantitative PCR (qRT-PCR) and Western blot respectively.

  Results In the model group, Beclin-1, LC3B, Bax and Caspase-3 genes were highly expressed at both the transcription and translation levels. Adding 10% Zuogui Pill containing serum or co culturing with membrane cells could down regulate the expression of these genes; The expression of p62 gene was lower than that of the control group. Adding 10% Zuogui Pill containing serum or co culturing with membrane cells could increase the expression of p62 gene in the model group. When 10% Zuogui Pill containing serum was added to the experimental group when co cultured with membrane cells, the expression of five genes changed more significantly.

  Conclusion Membrane cells can alleviate the chemotherapeutic injury of granulosa cells in vitro, and have a synergistic effect with Zuogui Pill containing serum, which is related to the inhibition of autophagy and the reduction of apoptosis of granulosa cells.