Experimental Study On Mechanism Of TCM Pingliukang In Treating Glioma

Object: This study was designed to observe inhibition of pingliukang on the profileration of C6 glioma cells in vitro by sero-pharmacological, and investigate the antineoplastic mechanism of pingliukang in cell cycle or cytokinetics.Methods:Part one: Through the sero-pharmacological method, cleaning wistar rats were divided into two sets randomly, and pingliukang were drenched to one group, 0.9% NS to the other. Then, the blood serum of rat was obtained.Then according to different concentration of serum, we divide the blood serum into four sets, 2.5%, 5%, 10% and 20%. C6 glioma cells were cultured with four different concentration of non-medicated and medicated serum respectively. We observed the growth status of C6 glioma cells by inverted microscope after 24hrs, 48hrs and 72hrs. Then, MTT was employed to measure the levels of the proliferation of C6 glioma cells cultured with different concentration of non-medicated and medicated serum at different times.Part two:According to part one, we found that the strongest inhibition happened in the of 10% and 20% concentration of medicated serum sets, and the most obvious inhibition happened in the 48hrs and the 72hrs stages. We select these two sets and started to culture the C6 glioma cells were, and collect enough cells to analyze the distribution of the cell cycles by FCM.Results:1. In the light microscope, typical features of apoptosis of tumor cell was showed in medicated serum sets, for instance, the cell volume grows downwards; cell nuclear turns solid and short; cytoplasm condenses; cellular membrane keeps integrity; chromatin agglutinates; cell losing microvillus; Cytoplasm pyknosis;and the cell organ gets dense. But in blank array and non-medicated serum arrays, the typical features of apoptosis rarely appeared.Along with the increasing concentration of serum, the inhibition of medicated serum on C6 increased.2. In the phase of 48hrs, when C6 glioma cells was cultured with serum concentration of 10%, comparing with non-medicated array, the G0/G1 period of medicated serum array changed little (P>0.05) , while the S period medicated serum array decreased (P<0.01) ; When C6 glioma cells was cultured with serum concentration of 20%, comparing with non-medicated serum, the G0/G1 period of medicated serum array increased (P<0.05) , while the S period decreased (P<0.01) .In the phase of 72h, when C6 glioma cells was cultured with serum concentration of 10%, comparing with non-medicated serum array, the G0/G1 period of medicated serum array increased (P<0.05) , but the S period decreased remarkably (P<0.01) . When concentration of serum increased to 20%, the cell cycle changed even more distinctly, the G0/G1 period of medicated serum array increased obviously(P<0.01), and the S period decreased obviously too( P<0.01).Conclusions:1. Composite pingliukang can inhibit proliferation of C6 glioma cells in vitro. The inhibitive effect shows relations of dose-dependent manner. T2. Composite pingliukang can disturb cell cycle of C6 glioma cell, block path of G0/G1 period to S period, and make G0/G1 period percentage increasing and S period decreasing. Then, composite pingliukang can reduce proliferation of C6 glioma cells.