酶解醇提辅助提取沙棘果渣黄酮及其光损伤修复活性研究

doi:10.3969/j.issn.2097-2806.2026.07.012

摘要/Abstract

摘要：

本研究针对沙棘果渣资源化利用瓶颈，优化黄酮类化合物提取工艺，探究沙棘黄酮对UVB诱导损伤HaCaT细胞的抗氧化保护作用。采用“脱脂-复合酶解-超声协同”技术：沙棘果渣经石油醚脱脂后，以2.5%果胶酶+2.5%纤维素酶破坏细胞壁，结合55%乙醇（含0.1 mol/L柠檬酸-磷酸盐缓冲液，pH 5.0，液料比（mL∶g）30∶1），在超声功率300 W、时间50 min、温度50 ℃条件下提取黄酮；构建150 mJ/cm² UVB损伤HaCaT细胞模型，设空白组（C）、模型组（UVB）及沙棘黄酮低/中/高剂量组（HRF-L/M/H，100，200，400 μg/mL），经沙棘黄酮处理后，通过MTT、EdU检测细胞活力与增殖，采用试剂盒法分析过氧化氢酶（CAT）、超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GPX）活性及活性氧（ROS）、丙二醛（MDA）水平。本研究优化工艺后的黄酮提取率为（9.92±0.44）mg/g；与C组相比，UVB组细胞活力、EdU阳性率降低，CAT、SOD、GPX活性下降，ROS、MDA升高（P<0.05）；与UVB组相比，HRF-L/M/H组细胞活力、EdU阳性率升高，CAT、SOD、GPX活性升高，ROS、MDA降低（P<0.05）。本研究表明，复合酶解-超声协同法可实现沙棘果渣黄酮高效提取；沙棘黄酮通过增强抗氧化酶活力、清除ROS、抑制脂质过氧化，缓解UVB对HaCaT细胞的损伤，本研究为沙棘果渣的资源利用及天然抗氧化剂开发提供了理论支撑。

关键词: 沙棘, 黄酮, 抗氧化, HaCaT细胞, 化妆品

Abstract:

This study aimed to address the bottleneck in the resource utilization of Hippophae rhamnoides Linn. pomace, optimize the extraction process, and investigate their antioxidant protective effects on HaCaT cells damaged by UVB irradiation. A “defatting-composite enzymatic hydrolysis-ultrasound synergistic” technique was adopted: After defatting Hippophae rhamnoides Linn. pomace with petroleum ether, a mixture of 2.5% pectinase and 2.5% cellulase was used to break the cell walls. Combined with 55% ethanol（containing 0.1 mol/L citrate-phosphate buffer, pH 5.0, liquid-to-solid ratio of 30∶1 mL/g）, flavonoids were extracted under ultrasonic conditions of 300 W, for 50 min, and at 50 ℃. A HaCaT cell model damaged by 150 mJ/cm² UVB was established, including a blank group（C）, a model group（UVB）, and Hippophae rhamnoides Linn. flavonoid low/middle/high dose groups（HRF-L/M/H, 100, 200, 400 μg/mL, respectively）. After treatment with Hippophae rhamnoides Linn. flavonoids, cell viability and proliferation were detected by MTT and EdU assays, and the activities of catalase（CAT）, superoxide dismutase（SOD）, and glutathione peroxidase（GPX）, as well as the levels of reactive oxygen species（ROS）and malondialdehyde（MDA）were analyzed using commercial kits. The extraction yield of flavonoids after process optimization in this study is（9.92±0.44）mg/g, which is higher than those reported in previous literatures. Compared with the C group, the cell viability and EdU positive rate in the UVB group decrease, the activities of CAT, SOD, and GPX decrease, and the levels of ROS and MDA increase（P<0.05）. Compared with the UVB group, the cell viability and EdU positive rate in the HRF-L/M/H groups increase, the activities of CAT, SOD, and GPX increase, and the levels of ROS and MDA decrease（P<0.05）. This study demonstrates that the composite enzymatic hydrolysis-ultrasound synergistic method can achieve efficient extraction of flavonoids from Hippophae rhamnoides Linn. pomace. Hippophae rhamnoides Linn. flavonoids alleviate UVB-induced damage to HaCaT cells by enhancing the activities of antioxidant enzymes, scavenging ROS, and inhibiting lipid peroxidation. This study provides theoretical support for the resource utilization of Hippophae rhamnoides Linn. pomace and the development of natural antioxidants.

Key words: Hippophae rhamnoides Linn., flavonoids, antioxidant, HaCaT cells, cosmetics

中图分类号:

TQ658

于波, 王金玲, 吕雅娟, 李艳红. 酶解醇提辅助提取沙棘果渣黄酮及其光损伤修复活性研究[J]. 日用化学工业（中英文）, 2026, 56(7): 935-942.

Bo Yu, Jinling Wang, Yajuan Lv, Yanhong Li. Study on enzymatic hydrolysis-ethanol assisted extraction of flavonoids from Hippophae rhamnoides Linn. pomace and their photodamage repair activity[J]. China Surfactant Detergent & Cosmetics, 2026, 56(7): 935-942.

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实验号	A 超声功率/W	B 超声时间/min	C 提取温度/℃	D 液料比/（mL∶g）	提取率/ （mg/g）
1	200	30	40	20∶1	8.25±0.42
2	200	50	50	30∶1	9.52±0.51
3	200	70	60	40∶1	8.86±0.45
4	300	30	50	40∶1	9.21±0.48
5	300	50	50	30∶1	9.85±0.54
6	300	70	40	20∶1	8.56±0.43
7	400	30	60	30∶1	9.42±0.50
8	400	50	40	40∶1	8.78±0.46
9	400	70	50	20∶1	8.12±0.41
K₁	8.88	8.96	8.53	8.31
K₂	9.21	9.38	9.58	9.54
K₃	8.77	9.52	9.03	8.31
R	0.82	0.65	1.05	1.23

分组	相对细胞活力/%	EdU阳性率/%
C	100.00±1.64	52.00±3.06
UVB	47.55±2.14^*	19.51±0.69^*
HRF-L	57.56±1.90^*#	26.44±2.68^*#
HRF-M	71.17±5.26^*#&	32.25±0.64^*#&
HRF-H	83.32±4.12^*#&△	41.03±4.34^*#&△
F	231.942	132.931
P	<0.001	<0.001

分组	CAT活性/ （U/（mg prot））	SOD活性/ （U/（mg prot））	GPX活性/ （U/（mg prot））	ROS相对荧光强度	MDA含量/ （nmol/（mg prot））
C	31.36±1.50	45.77±5.30	50.43±2.25	1.00±0.05	1.40±0.06
UVB	14.66±0.82^*	25.74±0.70^*	31.77±0.93^*	5.27±0.41^*	3.18±0.13^*
HRF-L	20.59±1.08^*#	33.05±2.99^*#	36.78±2.14^*#	4.01±0.21^*#	2.68±0.17^*#
HRF-M	24.70±0.83^*#&	36.26±1.89^*#&	40.05±1.42^*#&	3.06±0.17^*#&	2.44±0.10^*#&
HRF-H	28.92±2.85^*#&△	42.05±2.27^*#&△	43.90±2.13^*#&△	2.00±0.09^*#&△	2.11±0.16^*#&△
F	103.170	39.643	87.984	326.018	155.099
P	<0.001	<0.001	<0.001	<0.001	<0.001