玉米赤霉烯酮吸附剂对后备母猪子宫中LC3、PCNA分布和表达的影响

doi:10.3864/j.issn.0578-1752.2021.18.018

Abstract

Abstract:

【Objective】 It has been proved that zearalenone (ZEA) can activate estrogen sensitive gene through estrogen receptor in animals, resulting in reproductive toxicity, and affecting endometrial cell growth, oocyte maturation and follicular granulosa cell proliferation. The purpose of this experiment was to investigate the effects of ZEA adsorbent (zeolite + montmorillonite combinations) on distribution and expression of LC3 and PCNA in the uterus of gilts, and to discuss the detoxification effect of the new ZEA adsorbent from the perspective of histochemistry. 【Method】 A total of 48 healthy gilts with a body weight of 30±2.11 kg were randomly divided into 6 groups (n = 8): control group with basal diet, the ZEA group with basal diet + 1.008 mg∙kg^-1ZEA, the 0.1% ZEA adsorbent (ZEA 0.1) group with ZEA diet + 1.0 g∙kg^-1 new adsorbent, the 0.25% ZEA adsorbent (ZEA 0.25) group with ZEA diet + 2.5 g∙kg^-1 new adsorbent, the 0.5% ZEA adsorbent (ZEA 0.1) group with ZEA diet + 5.0 g∙kg^-1 new adsorbent, and the ZEA montmorillonite (ZEA+M) group with ZEA diet + 2.5 g∙kg^-1 montmorillonite. The preliminary trail period was 7 d, and the trial period was 21 d. 【Result】 ZEA increased the uterine organ index, and adding 0.25% and 0.5% adsorbents in the diet decreased the index of uterine organs obviously. The LC3 and PCNA positive cells were mainly distributed in the glandular epithelial cells, and the LC3 immunoreactivity of lumen epithelial cells were weaker than that of glandular epithelial cells. The immunoreactivity of LC3 in control group was stronger than that of ZEA group. The immunoreactivity and number of positive cells of LC3 in the ZEA new adsorbent group were significantly higher than those in the ZEA group, and there was a dose-dependent trend, but the effect of the ZEA-M group was increased slightly more than that of ZEA group. The results of PCNA in the luminal and glandular epithelium were contrary to that of LC3. The WB and qRT-PCR results also showed that 0.25% and 0.5% new adsorbents could promote the expression of LC3, increase autophagy, and decrease the PCNA immunoreactive reaction. These results indicated that ZEA inhibited the expression of LC3, inhibited autophagy and destroyed the homeostasis of endometrial cells and glandular epithelial cells. However, the new adsorbent could increase the expression of LC3 protein and decrease the expression of PCNA, which could promote autophagy and resist the abnormal proliferation of endometrial cells caused by ZEA. The new adsorbent had a good protective effect on uterine cells through bidirectional regulation.The experimental results provided theoretical basis for the further application of the new adsorbent. 【Conclusion】In this experiment, ZEA induced the uterus proliferation reaction, the new adsorbent (zeolite + montmorillonite combinations) resisted the negative effect of ZEA on the normal physiological function of uterus within a certain limits. As a result, 0.25% and 0.5% doses of this newadsorbents was suitable.

Key words: zearalenone, uterus, LC3, PCNA, gilts, adsorbent

HUANG LiBo,WANG JinQuan,GAO WenBo,CHEN HongJu,HOU YanMeng,YUAN XueJun,WANG ChunYang. Effects of Dietary Zearalenone Adsorbent on the Distribution and Expression of LC3 and PCNA in the Uterus of Gilts[J].Scientia Agricultura Sinica, 2021, 54(18): 4008-4017.

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日粮组成 Ingredients	含量 Content(%)	营养水平¹Nutrient levels
玉米Corn (%)	66.00	代谢能 Metabolic energy (MJ·kg^-1)	12.79
豆粕 Soybean meal (%)	21.00	粗蛋白 Crude proetin (%)	17.40
麸皮 Wheat bran (%)	5.00	钙 Calcium (%)	0.78
膨化大豆 Expanded soybean (%)	4.00	有效磷 Phosphorus (%)	0.30
预混料²Premix (%)	4.00	赖氨酸 Lysine (%)	0.96
合计 Total	100	蛋+胱 Sulphur amino acid (%)	0.60

目的基因 Target gene	引物序列(5' -3') Primer sequence	产物大小 Product size (bp)	登录号 Accession No.
GAPDH	F: ATGGTGAAGGTCGGAGTGAA	154	NM_001206359.1
GAPDH	R:CGTGGGTGGAATCATACTGG	154	NM_001206359.1
LC3	F: AACGAAATTCCTGGTGCCTGA	90	NM_100322893
LC3	R: :AAGGCTTGGTTAGCATTGAGCTG	90	NM_100322893
PCNA	F: GTGATTCCACCACCATGTTC	145	NM_001291925.1
PCNA	R: TGAGACGAGTCCATGCTCTG	145	NM_001291925.1

项目 Items	对照组Control	ZEA	ZEA0.1	ZEA0.25	ZEA0.5	ZEA-M
子宫器官指数 Organ index of uterus (g∙kg^-1)	1.91±0.31a	2.72±0.52b	2.05± 0.35a	1.93± 0.27a	1.84± 0.29a	1.81± 0.15a
数值标不同小写字母者表示差异显著(P<0.05)。n=8 Values with different superscripts differ significantly at P<0.05. n=8

Effects of Dietary Zearalenone Adsorbent on the Distribution and Expression of LC3 and PCNA in the Uterus of Gilts

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