枸杞多糖治疗雷公藤甲素诱导雄性小鼠生殖损伤的作用研究

doi:10.16431/j.cnki.1671-7236.2023.09.028

摘要/Abstract

摘要： 【目的】探究枸杞多糖(LBP)对雷公藤甲素诱导的雄性小鼠生殖损伤的恢复及治疗作用,为雷公藤甲素引起的生殖损伤提供临床基础治疗,为雷公藤甲素的广泛应用提供抗毒配伍药物。【方法】选取45只SPF级4周龄雄性ICR小鼠,采用数字法随机均分为5组:对照组(CON)、雷公藤甲素损伤组(TP)及枸杞多糖低(LBP-L)、中(LBP-M)、高(LBP-H)剂量组。小鼠除对照组腹腔注射生理盐水外,其余各组均腹腔注射120 μg/kg雷公藤甲素,每天1次;2 h后,对照组和雷公藤甲素损伤组灌胃蒸馏水,枸杞多糖低、中、高剂量组分别灌胃20、40和60 mg/kg枸杞多糖溶液,连续14 d后处死采样。利用精液分析仪检测精液质量,HE染色观察小鼠睾丸组织细胞病理变化;利用试剂盒检测小鼠睾丸组织超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性及丙二醛(MDA)、血清睾酮(T)含量;采用Western blotting检测小鼠睾丸组织B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、Cl-半胱氨酸蛋白酶-3(Cl-Caspase3)蛋白的表达情况。【结果】与对照组相比,雷公藤甲素损伤组小鼠体重增长率、脏器系数、精子数目、精子活率、SOD和GSH-Px活性均极显著下降(P＜0.01);精子畸形率和MDA含量均极显著上升(P＜0.01);睾丸组织病理学检测中生精细胞萎缩、精子细胞减少、细胞层次紊乱和空泡化结构严重。与雷公藤甲素损伤组相比,各剂量枸杞多糖组小鼠除血清睾酮外其余指标均有所改善(P＜0.05;P＜0.01),且枸杞多糖高剂量组改善效果最优。Western blotting结果表明,与对照组相比,枸杞多糖高剂量组小鼠睾丸组织中凋亡蛋白表达量显著或极显著升高(P＜0.05;P＜0.01);与雷公藤甲素损伤组相比,枸杞多糖高剂量组小鼠睾丸组织中Bax、Cl-Caspase3蛋白表达量和Bax/Bcl-2比值显著或极显著降低(P＜0.05;P＜0.01);Bcl-2蛋白表达量极显著升高(P＜0.01)。【结论】枸杞多糖对雷公藤甲素造成的生殖损伤有一定的治疗作用,其治疗作用主要体现在增加生物体抗氧化应激水平,提高精子质量,缓解小鼠睾丸组织空泡状细胞和改善细胞紊乱,通过调节相关凋亡蛋白表达抑制睾丸细胞的凋亡。

关键词: 枸杞多糖; 雷公藤甲素; 生殖损伤; 睾丸; 抗毒配伍

Abstract: 【Objective】 This study was aimed to investigate the recovery and therapeutic effect of Lycium barbarum polysaccharide (LBP) on reproductive injury induced by triptolide in male mice,so as to provide clinical basis for the treatment of reproductive injury caused by triptolide and antitoxic compatible drugs for the wide application of triptolide.【Method】 Forty-five SPF grade 4-week-old male ICR mice were randomly divided into 5 groups by digital method:Control group (CON),triptolide injury group (TP),Lycium barbarum polysaccharide low (LBP-L),medium (LBP-M) and high(LBP-H) dose treatment groups.Except for mice in control group were intraperitoneally injected physiological saline,mice in the other groups were intraperitoneally injected with 120 g/kg triptolide,once a day.Two hours later,the CON and TP groups were intragaically irrigated with distilled water,the LBP-L,LBP-M and LBP-H groups was intragaically irrigated with 20,40 and 60 mg/kg Lycium barbarum polysaccharide solution.Mice were killed after consecutive 14 days.Semen quality was measured by semen analyzer,HE staining sections were used to observe the pathological changes of testis in mice.The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px),and the contents of malondialdehyde (MDA) and serum testosterone (T) in testis of mice were detected by the kit.The expression of B cell lymphoma-2 (Bcl-2),Bcl-2 associated X protein (Bax) and Cl-Caspase3 protein in testis of mice were detected by Western blotting.【Result】 Compared with CON group,the growth rate of body weight,organ indexes,sperm number,sperm motility and the activities of SOD and GSH-Px in TP group were extremely significantly decreased (P＜0.01).The sperm malformation rate and MDA content were extremely significantly increased (P＜0.01).The results of testicular histopathology showed atrophy of spermatogenic cells,reduction of spermatogenic cells,disorder of cell hierarchy and serious vacuolation structure.Compared with TP group,other indexes except for serum testosterone were improved in different doses of Lycium barbarum polysaccharide groups (P＜0.05 or P＜0.01),and the high dose group had the best improvement effect.Western blotting results showed that compared with CON group,the expression of apoptosis proteins in testis of mice in LBP-H group were significantly or extremely significantly increased (P＜0.05 or P＜0.01).Compared with TP group,the expression of Bax and Cl-Caspase3 proteins and Bax/Bcl-2 ratio in testis of mice in LBP-H group were significantly or extremely significantly decreased (P＜0.05 or P＜0.01),the expression of Bcl-2 protein was significantly extremely increased (P＜0.01).【Conclusion】 Lycium barbarum polysaccharide had a therapeutic effect on the reproductive damage caused by triptolide,and its therapeutic effect was mainly reflected in the increase of antioxidant stress level of organisms,improving sperm quality,alleviating vacuolar cells and improving cell disorders in testis of mice,and inhibiting the apoptosis of testicular cells by regulating the expression of related apoptotic proteins.

Key words: Lycium barbarum polysaccharide; triptolide; reproductive injury; testis; antitoxin compatibility

中图分类号:

S859.8

王泽朝, 何文莹, 常青, 彭涛, 李艳宁, 毕逢辰, 王俊燕, 王银, 王丽娟. 枸杞多糖治疗雷公藤甲素诱导雄性小鼠生殖损伤的作用研究[J]. 中国畜牧兽医, 2023, 50(9): 3719-3729.

WANG Zechao, HE Wenying, CHANG Qing, PENG Tao, LI Yanning, BI Fengchen, WANG Junyan, WANG Yin, WANG Lijuan. Effect of Lycium barbarum Polysaccharide on Reproductive Injury Induced by Triptolide in Male Mice[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3719-3729.

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