茶籽壳发酵物的抗氧化活性及其对细胞内氧化损伤抑制作用

doi:10.3969/j.issn.0253-2417.2019.04.015

摘要/Abstract

摘要：

利用固定化细胞发酵技术，选取黑曲霉、枯草芽孢杆菌和酿酒酵母对茶籽壳进行发酵，制备发酵物。分析显示：与未经发酵的水提取物相比，茶籽壳酿酒酵母发酵物（FETSS-3）主要抗氧化活性物质含量明显增加，其多糖、多酚和皂苷含量分别增加至原来的9.19、9.59和4.67倍。从生化水平、细胞水平和酶活性水平对其抗氧化活性进行了探究，并对其皮肤安全性进行试验确认。研究结果表明：在0.1~0.4 g/L和0.5~2.0 g/L，FETSS-3的ABTS和·OH自由基清除活性显著优于V_C（P < 0.01；P < 0.01）；在0.2~1.2 g/L，其对脂质过氧化半抑制率IC₅₀仅为V_E的21.98%，抑制活性显著高于V_E（P < 0.01）。FETSS-3在20~120 mg/L能显著降低H₂O₂诱导的人皮肤成纤维细胞（HSF）内的氧化损伤应激水平（P < 0.01）。体外酶活性实验表明：FETSS-3（120 mg/L）能显著提高HSF细胞氧化应激损伤后的过氧化氢酶（CAT）、超氧化物歧化酶（SOD）和人谷胱甘肽-过氧化物酶（GSH-PX）活性（P < 0.05；P < 0.01；P < 0.01）。人体斑贴实验表明：FETSS-3无皮肤过敏风险。

关键词: 茶籽壳, 固定化细胞, 发酵, 抗氧化

Abstract:

Tea seed shell(TSS) were fermented with Aspergillus niger, Bacillus subtilis and Saccharomyces cerevisiae, by using immobilized cell fermentation technology, and the water extract of the fermentation product was prepared. The analysis showed that compared with the unfermented water extract, the content of main antioxidant substances in the fermented extract of S. cerevisiae (FETSS-3) increased significantly, the content of polysaccharides, polyphenols and saponins increased to 9.19, 9.59 and 4.67 times respectively. Its antioxidant activity was investigated by biochemical level, cellular level and enzyme activity level, and its skin safety was confirmed by patch test. The results showed that in the concentration range of 0.1-0.4 g/L and 0.5-2.0 g/L, ABTS and·OH radical scavenging activities of FETSS-3 were significantly superior to those of V_C(P < 0.01; P < 0.01); In the concentration range of 0.2-1.2 g/L, its IC₅₀ was only 21.98% of that of V_E, and its inhibitory activity was significantly higher than that of V_E(P < 0.01). In the concentration range of 20-120 mg/L, FETSS-3 could significantly reduce the level of oxidative damage stress of human skin fibroblasts(HSF) induced by H₂O₂(P < 0.01). In vitro enzyme activity experiments showed that FETSS-3(120 mg/L) could significantly improve the activity ofcatalase(CAT), superoxide dismutase(SOD) and glutathione peroxidase(GSH-PX) in HSF cells after oxidative stress injury(P < 0.05; P < 0.01; P < 0.01). The human body patch experiment showed that the FETSS-3 had no risk of skin allergy.

Key words: tea seed shell, immobilized cell, fermentation, antioxidation

中图分类号:

TQ35
TQ658.2

周晶,桂雨豪,龚盛昭. 茶籽壳发酵物的抗氧化活性及其对细胞内氧化损伤抑制作用[J]. 林产化学与工业, 2019, 39(4): 105-112.

Jing ZHOU,Yuhao GUI,Shengzhao GONG. Antioxidant Activity of Tea Seed Shell Fermentation Extracts and Its Inhibitory Effect on Intracellular Oxidative Damage[J]. Chemistry and Industry of Forest Products, 2019, 39(4): 105-112.

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样品¹⁾ samples	多糖类/(mg·g^-1) polysaccharides	多酚类/(mg·g^-1) polyphenols	皂苷类/(mg·g^-1) saponins
WETSS	16.25±0.89	14.96±0.54	23.28±0.59
FETSS-1	120.45±4.98^**	86.89±1.74^**	35.24±1.61
FETSS-2	59.42±2.91^*	48.49±0.86	66.54±1.08^*
FETSS-3	149.43±5.18^**	143.58±4.86^**	85.43±2.88^**

H₂O₂/(μmol·L^-1)	不同孵育时间阶段的MTT值MTT values at different incubation time stages
H₂O₂/(μmol·L^-1)	6 h	2 h	18 h	24 h
0	0.822±0.019	0.817±0.026	0.819±0.031	0.831±0.014
40	0.802±0.008^**	0.797±0.023^*	0.771±0.022	0.723±0.013^*
80	0.788±0.012^**	0.779±0.018^*	0.661±0.033^**	0.768±0.014^*
120	0.784±0.023^*	0.744±0.024^**	0.603±0.016^*	0.542±0.018^**
160	0.769±0.032	0.712±0.012^*	0.501±0.009^*	0.356±0.021
200	0.748±0.019^**	0.654±0.011^*	0.411±0.020^**	0.242±0.020^**
240	0.707±0.042	0.573±0.028^*	0.349±0.018^*	0.225±0.015
280	0.671±0.023	0.474±0.029^**	0.327±0.007^**	0.212±0.033^*
320	0.623±0.017^*	0.409±0.010	0.318±0.027^*	0.204±0.034^**
360	0.491±0.009	0.39±0.035^*	0.278±0.009	0.153±0.037^**

组名group	酶活力值enzyme activity value/(U·L^-1)¹⁾
组名group	CAT	GSH-PX	SOD
空白对照组the blank control group	(58.44± 8.17)×10³	28.46±5.023	461.49±21.38
H₂O₂损伤模型组H₂O₂ damage model group	(17.45±6.48)×10³ ^A	6.92±1.103 ^A	210.74±12.45 ^a
FETSS-3(40 mg/L)	(29.48±8.22)×10³ ^A	19.33±6.205 ^Ab	249.52±24.22 ^Ab
FETSS-3(80 mg/L)	(49.42±8.42)×10³ ^aB	26.54±6.158 ^b	340.71±27.64 ^ab
FETSS-3(120 mg/L)	(56.81±14.78)×10³ ^B	34.38±8.302 ^B	484.29±33.11 ^aB
FETSS-3(160 mg/L)	(58.93±11.72)×10³ ^B	35.38±7.603 ^aB	488.24±31.08 ^B

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