红景天多糖对低氧环境下猪睾丸间质细胞增殖及凋亡的影响

doi:10.11843/j.issn.0366-6964.2024.06.016

Abstract

Abstract:

This study investigates the effect and mechanism of rhodiola sachalinensis polysaccaride (RSP) on the proliferation and apoptosis of procine leydig cells under hypoxia conditions. The appropriate hypoxia injury time, optimal RSP concentration and the pre-treattime duration determined by CCK-8 assay. The experiment was divided into 3 groups, normal group, hypoxia group, and hypoxia+RSP group, and cell cycle proliferation and apoptosis were detected by flow cytometry method, the cell proliferation and apoptosis-related genes (CCND1, CDK4, IL-6, TNF-a, Bcl-2, Bax and Caspase-3) mRNA expression levels was detect by RT-qPCR, Western blot was used to detect the expression of Bax, Bcl-2, p21, JNK, p-JNK and p53. The viability of porcine leydig cells was decreased under hypoxia, which could be alleviated after 18 hours of pretreatment with 0.012 5 mg·mL^-1 RSP. The Flow cytometry results showed that RSP could alleviate cell cycle arrest induced by hypoxia in porcine leydig cells. RSP significantly up-regulated the expression of Bcl-2, CCND1, CDK4(P < 0.05), and down-regulated the expression of TNF-α, IL-6, Bax, Caspase-3, and p21, p53, and pJNK/JNK (P < 0.05). RSP helps ensure the normal progression of leydig cells from S-phase to G2-phase under hypoxia conditions, promote cell proliferation and inhibit apoptosis. It is speculated that RSP regulates cell proliferation and apoptosis through p53/p21 and JNK pathways.

Key words: RSP, porcine leydig cells, hypoxia, proliferation, apoptosis

CLC Number:

S828.3

Jinting LUO, Fafang XU, Lei WANG, Xuan LUO, Yuhong MA, Jianbo ZHANG, Weihua HUANG, Yuejun SHANG, Guofang WU. The Effect of RSP on Cell Proliferation and Apoptosis of Porcine Leydig Cells with Hypoxia[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(6): 2441-2450.

Figures/Tables 9

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基因 Gene	引物序列(5′→3′) Primers sequence	退火温度/℃ Tm
TNF-α	F: ACCACGCTCTTCTGCCTACTG	59.8
	R: TGAGACGATGATCTGAGTCCTTGG	58.2
IL-6	F: TTCAGTCCAGTCGCCTTCTCC	59.5
	R: TGGCATCACCTTTGGCATCTTC	58.2
CCND1	F: CGCCCTCCGTGTCCTACTTC	61.0
	R: GACCTCCTCCTCGCACTTCTG	60.4
CDK4	F: GTATGAACCAGTGGCGGAGATTG	58.3
	R: CCTCCAGCACCTCCTCCATTG	61.0
Bax	F: CATCTACCAAGAAGTTGAGCGAGTG	57.6
	R: ACGGCTGCGATCATCCTCT	59.3
Bcl-2	F: CGCAGAGGGGCTACGAGTG	61.7
	R: CGGGCTGGGAGGAGAAGATG	61.0
Caspase-3	F: GCTGTAGAACTCTAACTGGCAAACC	57.8
	R: CCCACTGTCCGTCTCAATCCC	60.8
β-actin	F: GATCTGGCACCACACCTTCTACAAC	60.0
	R: TCATCTTCTCACGGTTGGCTTTGG	59.7

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The Effect of RSP on Cell Proliferation and Apoptosis of Porcine Leydig Cells with Hypoxia

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