Effects of Proanthocyanidin on the Proliferation of Sheep Follicular Granulosa Cells in vitro

doi:10.13560/j.cnki.biotech.bull.1985.2021-1177

Abstract

Abstract:

The aim of this study is to investigate the effects of proanthocyanidin（PC）on the proliferation of sheep follicular granulosa cells in vitro. Sheep follicular granulosa cells（GCs）were cultured in complete medium with different concentrations of PC（20,30,40,50,60,70 and 80 μg/mL）for 24,48 and 72 h,respectively. The proliferation rate of GCs was determined by MTT method. And the mRNA abundances of cell cycle related genes p21,p27 and cell apoptosis-related gene caspase-8 were detected by RT-qPCR. The results showed that the proliferation rate of GCs was the highest when the concentration of PC was 50 μg/mL when cultured for 48 h in vitro. And the relative mRNA abundances of three genes（p21,p27 and caspase-8）significantly decreased,respectively（P<0.01）. In sum,PC could promote the proliferation of sheep follicular GCs in vitro,and which was verified at relevant mRNA level.

Key words: proanthocyanidins, granulosa cells, sheep, in vitro, proliferation

YANG Xiao-feng, QIN Xiao-wei, GUO Ze-yuan, LV Li-hua. Effects of Proanthocyanidin on the Proliferation of Sheep Follicular Granulosa Cells in vitro[J]. Biotechnology Bulletin, 2022, 38(9): 258-263.

Figures/Tables 4

Table 1 Primer sequences of RT-qPCR

基因名称Gene name	引物序列 Primer sequence（5'-3'）
p21	F：ACGTCTCAGGAGGACCACTT, R：TGGTAGAAATCTGTCATGCTGGT
p27	F：TACTTGGGTCTCAGGCAAAC, R：GCTCTTTTGTTTTGAGGAGAGG
caspase-8	F：ACTGTGTGGAGCAGGTAACA, R：CCGGCTTAGGAACTTGAGGG
18S	F：GTAACCCGTTGAACCCCATT, R：CCATCCAATCGGTAGTAGCG

Fig. 1 Growth status of sheep follicular granulosa cells in vitro cultured with different concentrations of PC for different time（100×） A1：48 h,0 μg/mL;B1：48 h,20 μg/mL;B2：48 h,50 μg/mL;B3：48 h,70 μg/mL;A2：24 h, 0 μg/mL;A3：72 h, 0 μg/mL;C：24 h, 50 μg/mL;D：72 h, 50 μg/mL

Fig. 2 OD values of granulosa cells cultured in vitro after 24,48 and 72 h treatment with different concentration of PC Superscript lowercase letters indicate comparison results at 0.05 significance level

Fig. 3 mRNA expression of proliferation-related genes in sheep granulosa cells Superscript capital letters indicate comparison results at 0.01 significance level

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