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

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (6): 2441-2450.doi: 10.11843/j.issn.0366-6964.2024.06.016

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

罗金婷¹(), 许发芳², 王磊¹, 罗璇¹, 马玉红¹, 张剑搏¹, 黄伟华¹, 尚月军³, 吴国芳¹^,*()

1. 青海大学畜牧兽医科学院，青海省高原家畜遗传资源保护与创新利用重点实验室，农业农村部高原畜禽遗传育种重点实验室，西宁 810016
2. 青海省互助八眉猪原种育繁场，海东 810599
3. 青海省农业农村科技指导发展服务中心，西宁 810008

收稿日期:2023-12-01 出版日期:2024-06-23 发布日期:2024-06-28
通讯作者: 吴国芳 E-mail:1356372949@qq.com;qhdxwuguofang@126.com
作者简介:罗金婷(1998-)，女，广东茂名人，硕士生，主要从事畜牧方面的研究，E-mail: 1356372949@qq.com
基金资助:
青海省自然基金应用基础研究(2022-ZJ-752)

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

Jinting LUO¹(), Fafang XU², Lei WANG¹, Xuan LUO¹, Yuhong MA¹, Jianbo ZHANG¹, Weihua HUANG¹, Yuejun SHANG³, Guofang WU¹^,*()

1. Key Laboratory of Plateau Livestock Genetic Breeding of Ministry of Agriculture and Rural Affairs, Key Laboratory of Plateau Livestock Genetic Resources Protection and Innovative Utilization of Qinghai Province, Qinghai Academy of Animal Science and Veterinary Medicine, Qinghai University, Xining 810016, China
2. Qinghai Mutual Aid Bamei Pig Breeding Field, Haidong 810599, China
3. Qinghai Province Agriculture and Rural Science and Technology Guidance Development Service Center, Xining 810008, China

Received:2023-12-01 Online:2024-06-23 Published:2024-06-28
Contact: Guofang WU E-mail:1356372949@qq.com;qhdxwuguofang@126.com

摘要/Abstract

摘要：

本研究探讨了红景天多糖(rhodiola sachalinensis polysaccaride，RSP)对缺氧条件下猪睾丸间质细胞增殖和凋亡的影响及其机制。通过CCK-8法测定细胞活力，筛选出RSP的最适宜浓度和预处理时间；设正常组、低氧组、低氧+RSP组，流式细胞术检测细胞周期和凋亡率，RT-qPCR检测细胞增殖和凋亡相关基因(CCND1、CDK4、IL-6、TNF-a、Bcl-2、Bax、Caspase-3) mRNA表达水平，Western blot检测Bax、Bcl-2、p21、JNK、p-JNK、p53表达水平。低氧条件下，猪睾丸间质细胞活力下降，经0.012 5 mg·mL^-1 RSP预处理18 h后，细胞活力有所提高。流式细胞术结果显示，RSP能缓解猪睾丸间质细胞低氧引起的细胞周期阻滞。RSP显著上调Bcl-2、CCND1、CDK4的表达(P＜0.05)，下调TNF-α、IL-6、Bax、Caspase-3、p21、p53、pJNK/JNK的表达(P＜0.05)。RSP能保证低氧环境下猪睾丸间质细胞由S期进入G2期，促进细胞增殖，抑制细胞凋亡，推测RSP通过p53/p21和JNK通路调控细胞增殖和凋亡。

关键词: 红景天多糖, 睾丸间质细胞, 低氧环境, 增殖, 凋亡

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

中图分类号:

S828.3

罗金婷, 许发芳, 王磊, 罗璇, 马玉红, 张剑搏, 黄伟华, 尚月军, 吴国芳. 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[J]. 畜牧兽医学报, 2024, 55(6): 2441-2450.

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.

图/表 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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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

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

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