牡蛎酶解物减轻5-FU对荷瘤小鼠肠黏膜损伤的作用研究*

doi:10.11978/2021155

热带海洋学报 ›› 2022, Vol. 41 ›› Issue (5): 105-116.doi: 10.11978/2021155CSTR: 32234.14.2021155

牡蛎酶解物减轻5-FU对荷瘤小鼠肠黏膜损伤的作用研究*

叶子晴¹^,²^,⁴(), 赵祥弹¹^,²^,⁴, 蔡冰娜¹^,³^,⁴, 万鹏¹^,³^,⁴, 陈华¹^,³^,⁴, 潘剑宇¹^,³^,⁴()

1.中国科学院南海海洋研究所, 中国科学院热带海洋生物资源与生态重点实验室, 广东广州 510301
2.中国科学院大学, 北京 100049
3.南方海洋科学与工程广东省实验室(广州), 广东广州 511458
4.中国科学院南海生态环境工程创新研究院, 广东广州 510301

收稿日期:2021-11-10 修回日期:2021-12-21 出版日期:2022-09-10 发布日期:2021-12-22
通讯作者: 潘剑宇
作者简介:叶子晴(1996—), 女, 广东省云浮市人, 硕士研究生, 从事海洋生物活性肽的分离纯化与活性研究。email: yeziqing19@mails.ucas.ac.cn
基金资助:
国家重点研发计划(2018YFC0311202);广东省海洋经济发展专项(GDNRC[2020]038);广东省海洋经济发展专项(GDNRC[2020]036);广东省自然科学基金(2018A030313088);南方海洋科学与工程广东省实验室(广州)人才团队引进重大专项(GML2019ZD0406);广东省重点领域研发计划(2020B1111030004);中国科学院南海生态环境工程创新研究院自主部署项目(ISEE2021PY05)

Oyster hydrolysates alleviate 5-fluorouracil-induced intestinal mucosal injury on S180 tumor-bearing mice*

YE Ziqing¹^,²^,⁴(), ZHAO Xiangdan¹^,²^,⁴, CAI Bingna¹^,³^,⁴, WAN Peng¹^,³^,⁴, CHEN Hua¹^,³^,⁴, PAN Jianyu¹^,³^,⁴()

1. CAS Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China
2. University of Chinese Academy of Sciences, Beijing 100049, China
3. Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou), Guangzhou 511458, China
4. Institution of South China Sea Ecology and Environmental Engineering, Chinese Academy of Sciences, Guangzhou 510301, China

Received:2021-11-10 Revised:2021-12-21 Online:2022-09-10 Published:2021-12-22
Contact: PAN Jianyu
Supported by:
National Key Research and Development Program of China(2018YFC0311202);Marine Economy Development Special Project of Guangdong Province(GDNRC[2020]038);Marine Economy Development Special Project of Guangdong Province(GDNRC[2020]036);National Natural Science Foundation of Guangdong, China(2018A030313088);Key Special Project for Introduced Talents Team of Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou)(GML2019ZD0406);Key-Area Research and Development Program of Guangdong Province(2020B1111030004);Innovation Academy of South China Sea Ecology and Environmental Engineering, Chinese Academy of Sciences(ISEE2021PY05)

摘要/Abstract

摘要：

文章以牡蛎肉为原料, 采用胰蛋白酶水解法制备获得牡蛎胰蛋白酶水解物(trypsin hydrolysates of oyster, OYH), 分析了其氨基酸组成和相对分子量分布, 并初步探究了OYH在减轻化疗药物5-氟尿嘧啶(5-fluorouracil, 5-FU)对S180荷瘤小鼠肠黏膜损伤方面的作用效果。结果表明: OYH中的必需氨基酸占总氨基酸的36.44%, 且谷氨酸含量最高, 其分子质量主要集中在1kDa以下, 所占比例为92.18%。5-FU单独使用组的肿瘤抑制率为31.00%; 与模型组相比, OYH-L+5-FU组、OYH-M+5-FU组和OYH-H+5-FU组小鼠肿瘤抑制率明显提高(p<0.001), 分别为38.24%、46.27%和47.57%, 其中OYH-M+5-FU组和OYH-H+5-FU组较5-FU单独使用有明显提高(p<0.01, p<0.001)。同时, OYH还能有效减轻5-FU给药期间造成的肠道黏膜损伤, 与5-FU组相比, OYH-L+5-FU组、OYH-M+5-FU组和OYH-H+5-FU组小鼠的小肠绒毛高度、隐窝深度、黏膜厚度和绒毛表面积均显著增加(p<0.01, p<0.001), 明显改善小肠隐窝和上皮绒毛的状态, 维持肠黏膜屏障的完整。此外, OYH-L+5-FU组、OYH-M+5-FU组和OYH-H+5-FU组的S180荷瘤小鼠与模型组相比, 中性粒细胞含量降低, 淋巴细胞含量增加(p<0.05, p<0.01), OYH-H+5-FU组能显著增加机体总蛋白、白蛋白以及免疫球蛋白含量(p<0.05, p<0.01, p<0.001), 改善机体免疫功能、营养状况以及调节血清免疫抗体水平。结果显示牡蛎肉的胰蛋白酶水解物具有作为化疗过程辅助治疗营养食品, 起到肠黏膜保护和提升机体耐受力的应用潜力。

关键词: 牡蛎酶解物, 化疗, 肿瘤抑制, 肠黏膜保护, 免疫调节

Abstract:

In this study, oyster meat was used as the raw material to produce trypsin hydrolysates of oyster (OYH). The amino acid composition and relative molecular weight distribution of OYH were then analyzed. The effect of OYH on reducing 5-fluorouracil (5-FU) induced intestinal mucosal injury on S180 tumor-bearing mice was preliminarily investigated. The results showed that the essential amino acids in the total amino acids of OYH accounted for 36.44 %, and content of glutamic acid was the highest. The molecular weight of OYH was mostly below 1 kDa, accounting for 92.18 %. The results showed that the tumor inhibition ratio of 5-FU group was 31.00 %. Compared with model group, the tumor inhibition ratio of mice in OYH-L+5-FU, OYH-M+5-FU and OYH-H+5-FU group were significantly increased (p < 0.001) with the percentage of 38.24 %, 46.27 % and 47.57 %, respectively. Tumor inhibition ratios of OYH-M+5-FU and OYH-H+5-FU group were greatly higher than 5-FU (p < 0.01, p < 0.001). Meanwhile, OYH can alleviate intestinal mucosal injury caused by 5-FU chemotherapy. Compared with 5-FU alone treatment, OYH-L+5-FU, OYH-M+5-FU and OYH-H+5-FU group significantly increased the villus height, crypt depth, mucosa thickness and villus surface area of small intestine (p < 0.01, p < 0.001), which indicated that OYH can improve the state of intestinal crypt and epithelial villi, maintain the integrity of the intestinal mucosal barrier during chemotherapy. In addition, compared with model group, the percentage of neutrophils was decreased but the percentage of lymphocytes was increased in OYH-L+5-FU, OYH-M+5-FU and OYH-H+5-FU group (p < 0.05, p < 0.01). And the OYH-H+5-FU group can largely increase the levels of total protein, albumin and immune globulin in serum (p < 0.05, p < 0.01, p < 0.001), which showed that OYH play a role in the immunologic function, nutritional improvement and level of immune antibody regulation in serum immunoregulation. In conclusion, trypsin hydrolysates of oyster meat have the potential for becoming an adjuvant nutritional food in the process of chemotherapy, protecting intestinal mucosa and improving body tolerance.

Key words: oyster hydrolysates, chemotherapy, anti-tumor, intestinal mucosal protection, immune regulation

中图分类号:

P735.4

叶子晴, 赵祥弹, 蔡冰娜, 万鹏, 陈华, 潘剑宇. 牡蛎酶解物减轻5-FU对荷瘤小鼠肠黏膜损伤的作用研究*[J]. 热带海洋学报, 2022, 41(5): 105-116.

YE Ziqing, ZHAO Xiangdan, CAI Bingna, WAN Peng, CHEN Hua, PAN Jianyu. Oyster hydrolysates alleviate 5-fluorouracil-induced intestinal mucosal injury on S180 tumor-bearing mice*[J]. Journal of Tropical Oceanography, 2022, 41(5): 105-116.

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组名	是否移植肉瘤细胞株	腹腔注射5-氟尿嘧啶剂量/(mg·kg^-1)	每日灌胃牡蛎酶解物剂量/(mg·kg^-1)
正常组	否	—	—
模型组	是	—	—
5-氟尿嘧啶组(5-FU组)	是	30	—
低剂量牡蛎胰蛋白酶水解物联合5-氟尿嘧啶组(OYH-L+5-FU组)	是	30	200
中剂量牡蛎胰蛋白酶水解物联合5-氟尿嘧啶组(OYH-M+5-FU组)	是	30	400
高剂量牡蛎胰蛋白酶水解物联合5-氟尿嘧啶组(OYH-H+5-FU组)	是	30	800

氨基酸种类	氨基酸含量/%	氨基酸种类	氨基酸含量/%
天冬氨酸 Asp	3.76	亮氨酸 Leu*	2.39
苏氨酸 Thr*	1.65	酪氨酸 Tyr	1.60
丝氨酸 Ser	1.44	苯丙氨酸 Phe*	1.27
谷氨酸 Glu	5.80	赖氨酸 Lys*	2.58
脯氨酸 Pro	1.99	组氨酸 His*	0.88
甘氨酸 Gly	3.08	精氨酸 Arg	2.30
丙氨酸 Ala	2.46	色氨酸Trp*	0.23
缬氨酸 Val*	1.69	氨基酸总量TAA	35.29
蛋氨酸 Met*	0.60	必需氨基酸含量EAA	12.86
异亮氨酸 Ile*	1.57	EAA/TAA	36.44

组别	初体重/g	终体重/g	瘤重/g	抑瘤率/%
正常组	25.99±1.23	29.11±1.50	—	—
模型组	24.27±1.76	28.72±2.51	1.57±0.30	—
5-FU组	24.14±0.82	25.15±1.41^***△△△	1.09±0.16^***	31.00
OYH-L+5-FU组	24.58±1.41	25.95±1.45^**△△△	0.97±0.18^***	38.24
OYH-M+5-FU组	24.61±2.13	26.64±1.65^*#△△	0.85±0.09^***###	46.27
OYH-H+5-FU组	23.94±1.92	26.84±1.83^#△△	0.82±0.19^***##	47.57

	正常组	模型组	5-FU组	OYH-L+5-FU组	OYH-M+5-FU组	OYH-H+5-FU组
白细胞/(×10⁹L^-1)	6.61±2.58^***	13.22±3.23	4.71±0.99^***	4.25±1.14^***	4.12±0.46^***	4.40±0.81^***
红细胞/(×10¹²L^-1)	10.34±0.46^***	9.03±0.37	8.82±0.44	9.12±0.41	8.70±0.35^$	8.72±0.49
血红蛋白/(g·L^-1)	158.90±6.79^***	136.20±6.16	134.70±5.56	138.80±5.12	133.00±5.42^$	133.10±7.46
红细胞压积/%	44.93±1.68^***	39.13±1.27	38.53±1.60	39.25±1.40	37.73±1.51^*$	37.68±1.99
血小板/(×10⁹L^-1)	1312.30±149.95^***	1740.90±169.84	1601.50±196.78	1747.00±257.28	1640.40±170.42	1817.30±209.85^#
淋巴细胞/%	80.29±4.17^***	54.82±6.35	62.65±11.25	66.71±8.16^**	65.15±6.27^**	65.05±8.91^**
中性粒细胞/%	17.47±4.08^***	39.43±6.90	32.14±9.61	28.57±7.54^**	30.95±6.22^**	30.22±8.20^*
单核细胞/%	0.87±0.40^***	4.98±1.57	4.22±2.22	3.60±0.66^*	2.99±1.00^**	3.77±1.11
嗜酸性粒细胞/%	1.34±0.45^**	0.74±0.19	0.99±0.51	1.10±0.46^*	0.91±0.48	0.89±0.57
嗜碱性粒细胞/%	0.03±0.07	0.03±0.05	0.00±0.00	0.02±0.06	0.00±0.00	0.07±0.12

	正常组	模型组	5-FU组	OYH-L+5-FU组	OYH-M+5-FU组	OYH-H+5-FU组
总蛋白/(g·L^-1)	73.83±7.65^***	57.35±7.11	54.72±6.55	61.39±6.85^#	64.36±8.95^#	70.56±8.85^**###$
白蛋白/(g·L^-1)	48.41±4.62^***	36.15±5.41	29.90±3.23^**	37.25±8.01^#	38.36±9.21^#	44.58±5.75^**###$
前白蛋白/(mg·L^-1)	3.74±0.70^**	2.71±0.53	2.45±0.46	2.45±0.53	2.47±0.52	2.57±0.52
转铁蛋白/(g·L^-1)	10.31±1.54^**	8.52±1.08	8.64±1.21	9.25±1.06	9.42±1.18	9.32±1.00
补体C3 /(g·L^-1)	2.56±0.61^***	3.78±0.41	3.09±1.12	3.70±0.52	3.84±0.79	3.56±0.88
补体C4 /(g·L^-1)	0.38±0.07^***	0.53±0.06	0.40±0.08^**	0.43±0.09^*	0.43±0.07^**	0.43±0.08^**
免疫球蛋白A /(g·L^-1)	0.37±0.12^***	0.13±0.03	0.12±0.03	0.15±0.03^#	0.17±0.03^*##	0.18±0.05^*##
免疫球蛋白G /(g·L^-1)	13.54±1.98^**	10.57±2.06	10.12±0.98	10.71±2.39	11.47±2.36	11.83±1.77^#
免疫球蛋白M /(g·L^-1)	1.72±0.33^***	1.08±0.19	0.99±0.15	1.23±0.32^#	1.56±0.26^***###$	1.64±0.25^***###$$

牡蛎酶解物减轻5-FU对荷瘤小鼠肠黏膜损伤的作用研究*

Oyster hydrolysates alleviate 5-fluorouracil-induced intestinal mucosal injury on S180 tumor-bearing mice*

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