Objective: To investigate the effect of Liuwei Dihuangwan on
connexin 32 (Cx32) in
hepatoma cell line CBRH7919 and its
gap junction intercellular
communication (GJIC), and furthermore study its mechanism of enhancing the bystander
killing effect of
suicide gene therapy .
Method: Liuwei Dihuangwan (32 g·kg·d-1) and the same volume of
normal saline were given to the
rats by intragastrical
administration .
Blood was taken to prepare the medicated
serum of Liuwei Dihuangwan and blank control
serum , respectively. The
hepatoma cell line CBRH7919 were treated by control
serum and medicated
serum of Liuwei Dihuangwan in different concentrations. There were four groups in experimentthe blank
control group (volume fraction of 10%), medicated
serum high
dose group of Liuwei Dihuangwan (the volume fraction of 10%), medicated
serum middle
dose group of Liuwei Dihuangwan (the volume fraction of 5%), and medicated
serum low
dose group of Liuwei Dihuangwan (the volume fraction of 2.5%). The expression levels of
Cx32 protein and
mRNA in
hepatoma cell line CBRH7919 were detected by
indirect immunofluorescence assay (ⅡA) and Real-
time fluorescence quantitative
polymerase chain reaction (
Real-time PCR ) assay. The
fluorescence redistribution after
photobleaching (FRAP)
method was used to detect the function of GJIC of
hepatoma cell line CBRH7919.
Result: ① The
indirect immunofluorescence assay (ⅡA)
analysis indicated that as compared with the blank
control group , the cx32 expression of CBRH7919
cells was up-regulated in a concentration-dependent manner in each
dose group of the
serum containing Liuwei Dihuangwan (PPPP
Conclusion: The mechanism of medicated
serum of Liuwei Dihuangwan in enhancing the bystander
killing effect of
suicide geneis related to
gap junction . Liuwei Dihuangwan may enhance the function of GJIC by increasing the
localization of cx32 on the
cell membrane of CBRH7919
cells and increasing the expression of cx32
mRNA and
protein to achieve the synergistic action.