WANG Xiao-li, CUI Hong-xia, LI Hong-ling, et al. Effect of Tianma Gouteng Yin-containing Serum on 1-Methyl-4-phenylpyridinium (MPP)-induced PC12 Cell Apoptosis JNK Pathway[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(21): 141-146.
DOI:
WANG Xiao-li, CUI Hong-xia, LI Hong-ling, et al. Effect of Tianma Gouteng Yin-containing Serum on 1-Methyl-4-phenylpyridinium (MPP)-induced PC12 Cell Apoptosis JNK Pathway[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(21): 141-146. DOI: 10.13422/j.cnki.syfjx.2017210141.
Effect of Tianma Gouteng Yin-containing Serum on 1-Methyl-4-phenylpyridinium (MPP)-induced PC12 Cell Apoptosis JNK Pathway
Objective: To examine the effect of Tianma Gouteng Yin-containing serum on 1-methyl-4-phenylpyridinium (MPP+)-induced apoptosis of PC12 cells
and explore the neuroprotective effect of c-Jun N-terminal kinase (JNK) signaling pathway. Method: A total of 32 male SD rats were randomly divided into control group and high
middle and low-dose Tianma Gouteng Yin-containing serum groups (22.8
11.4
5.7 g·kg-1). The rats were intragastrically given Tianma Gouteng Yin to prepare Tianma Gouteng Yin-containing serum. The control group was given the same volume of normal saline. The proliferation of PC12 cells was cultured and divided into the blank group
the model group
and high
middle and low-dose Tianma Gouteng Yin-containing serum groups. The blank group and the model group were given blank serum
while the other three groups were given 10% Tianma Gouteng Yin-containing serum incubated for 30 min. The model group and Tianma Gouteng Yin-containing serum groups were additionally given 500 μmol·L-1 MPP+ and co-cultured for 48 h. The cell viability was measured by Cell Titer 96° Aoueous MTS Reagent powder (MTS). The follow-up experiment was divided into five groups
namely blank group
MPP+ group
MPP++high-dose Tianma Gouteng Yin-containing serum group
MPP++SP600125 group
and MPP++SP600125+high-dose Tianma Gouteng Yin-containing serum group. Apoptosis was analyzed by flow cytometry. Cysteine aspartate protease-3 (Caspase-3) activity was detected by spectrophotometry. The expressions of JNK
p-JNK
c-Jun
p-c-Jun protein were detected by Western blot. Result: Compared with the blank control
cell viability was significantly decreased in the model group (P < 0.05). Compared with the model group
cell viability was significantly increased in high
middle and low-dose Tianma Gouteng Yin-containing serum groups (P < 0.05). In the follow-up experiment
compared with blank control
apoptosis
Caspase-3 activity
and expressions of p-JNK and p-c-Jun protein were significantly increased in the MPP+ group (P < 0.05). Compared with the MPP+ group
poptosis
Caspase-3 activity
and expressions of p-JNK and p-c-Jun protein were significantly decreased in the MPP++high-dose Tianma Gouteng Yin-containing serum group and MPP++SP600125 group (P < 0.05). Co-treatment with SP600125 and high-dose Tianma Gouteng Yin-containing serum resulted in a slight increase in neuroprotection
which was significantly greater than high-dose Tianma Gouteng Yin-containing serum treatment alone (P < 0.05)
but not significantly greater than SP600125 treatment alone. The expressions of JNK and c-Jun protein showed no significant change. Conclusion: Tianma Gouteng Yin-containing serum has the neuroprotective effect against MPP+-induced apoptosis
which may be closely related to the JNK signaling pathway.
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Related Institution
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