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With the improvement of people's living standards, the incidence of diabetes is increasing worldwide, and it has become the third largest disease in human beings after cardiovascular and cerebrovascular diseases and cancer, which has attracted widespread attention
.
There are various causes of diabetes, among which the dysfunction of pancreatic islets β cells is the key cause
of diabetes.
A large number of previous studies have proved that isoflavones can inhibit the apoptosis of islet β cells, improve insulin secretion, and enhance the function of islet β cells, which is of great significance
for the prevention of metabolic syndrome such as diabetes.
The content of isoflavones in taro is as high as 41.
59 mg/g, which is an excellent source
of isoflavones.
Isoflavones have antioxidant, anticancer, estrogen-like, osteoporosis improvement, prevention of metabolic syndrome and other effects
.
Deng Ke, Chen Wei, Ni Qinxue* from the College of Food and Health of Zhejiang A&F University explored the enrichment process of potato isoflavone macroporous resin on the basis of tarato alcohol extract and ethyl acetate extract, and analyzed the antioxidant activity, α-glucosidase inhibition ability and protective effect of RIN-m5F cells on the basis of taro alcohol extract and ethyl acetate extract, in order to provide a theoretical basis
for the subsequent development of functional foods.
1.
Optimization of macroporous resin enrichment process of isoflavones
Screening of resin models
It can be seen from Figure 1A that among the six macroporous resins, the adsorption rate of D101 and AB-8 is higher than that of the other four resins.
The desorption rate of D101 and DM130 is better than that of the other four resins
.
D101 was superior to the other five macroporous resins in terms of adsorption and desorption, so D101 resin was selected to enrich isoflavones
.
Effect of sample mass concentration on adsorption rate
As shown in Figure 1B, when the mass concentration of the sample increases, the adsorption rate also gradually increases.
When the mass concentration was 1.
5 mg/mL, the adsorption rate reached the maximum value (83.
48%).
The resin adsorption state reaches saturation, and the mass concentration of the sample continues to increase, and the adsorption rate gradually decreases
.
Therefore, the optimal mass concentration is 1.
5 mg/mL
.
Effect of ethanol volume fraction on desorption
As shown in Figure 1C, as the volume fraction of ethanol increases, the desorption rate gradually increases; When the ethanol volume fraction reached 80%, the desorption rate reached the maximum (80.
61%); As the ethanol volume fraction continues to increase, the desorption rate begins to decrease and eventually flattens
.
This may be related to the polarity of the eluent, and large or small polarity is not conducive to the elution of isoflavones, so the optimal eluent ethanol volume fraction is 80%.
Effect of elution volume on desorption
It can be seen from Figure 1D that the desorption rate increases first and then decreases with the increase of the elution volume of 80% ethanol solution and a flow rate of 1.
0 mL/min
.
At an elution volume of 80 mL, the desorption rate reached a maximum (84.
63%)
.
When the elution volume is 120 mL, the effluent is basically colorless, indicating that the isoflavones adsorbed on the D101 resin have basically been eluted, so the optimal elution volume is 80 mL (4 column volumes).
Effect of eluent pH on desorption
It can be seen from Figure 1E that when the pH < 6, the desorption rate gradually increases with the increase of pH value.
At pH 6, the desorption rate reaches a maximum (80.
14%); The pH value continues to increase, and the adsorption rate gradually decreases<b10>.
Therefore, the pH value of the optimal eluent is 6
.
Too high or too low pH is not conducive to the adsorption and desorption of flavonoids, because the form of flavonoids changes with the change of pH, or the formation of salts, or the molecule loses the H+ on the hydroxyl group in the form of negative ions, and the two modes of existence are not conducive to the adsorption
of macroporous resins.
2.
AI-3 antioxidant activity and α-glucosidase inhibition
The antioxidant activity of AEx, AEx-EtOAc and AI-3 (IC50 and FRAP for DPPH radicals, ABTS cationic radicals) and α-glucosidase inhibitory capacity are shown in Table 1
.
3.
Protective effect of AI-3 on oxidative damage of RIN-m5F cells
Establishment of oxidative stress model
As shown in Figure 2, 50~300 μmol/L H2O2 damaged cells for 1 h, and the cell viability decreased significantly (P<0.
05).
<b10> The apoptosis rate of 150 and 200 μmol/L H2O2 cells was close to 50%, and combined with the experimental results and cell growth status, 150 μmol/L H2O2 treated cells were used for 1 h to establish an oxidative stress model
of RIN-m5F cells.
Effect of AI-3 on proliferative viability of RIN-m5F cells
As shown in Figure 3, the survival rate of RIN-m5F cells after 0~800 μg/mL AEx treatment for 24 h was not significantly different from that of the control group (P>0.
05), while the survival rate of RIN-m5F cells (81.
58%) after 1 600 μg/mL AEx treatment for 24 h was significantly lower than that of the control group (P<0.
05), indicating that the safe mass concentration<b10> of AEx was at 0~800 μg/mL 。 The survival rates of RIN-m5F cells in the mass concentration range of 0~400 and 0~300 μg/mL of AEx-EtOAc and AI-3 were not significantly different from those of the control group (P>0.
05), indicating that the safe mass concentrations of AEx-EtOAc and AI-3 were 0~400 and 0~300 μg/mL
, respectively.
Protective effect of AI-3 on oxidative damage in RIN-m5F cells
As shown in Figure 4, the effect of AEx on the survival rate of RIN-m5F cells in the range of safe action mass concentration (0~800 μg/mL) was not significantly different from that of H2O2 injury group (P>0.
05), and the effect of AEx-EtOAc on the survival rate of RIN-m5F cells in the range of safe action mass concentration (0~800 μg/mL) was very different from that of H2O2 injury group (P<0.
01); 25~300 μg/mL The cell survival rate of AI-3 group was significantly higher than that of H2O2 injury group (P<0.
01), and when AI-3 reached the upper limit of safe mass concentration (300 μg/mL), the cell survival rate reached the maximum, which was 84.
19%, which was higher than that of the positive control group<b10>.
conclusion
In summary, under optimized conditions, D101 macroporous resin can effectively enrich isoflavones, and the resulting component AI-3 has strong antioxidant activity, α-glucosidase inhibition and RIN-m5F cell oxidative damage protection
.
The research results can provide a theoretical basis
for the development of functional foods for the prevention of metabolic syndrome such as diabetes.
About the corresponding author
Ni Qinxue Associate Professor, College of
Food and Health, Zhejiang A&F University.
In 2004, he received a bachelor's degree in agronomy from Zhejiang A&F University; Master of Agronomy, Kyungpook National University, South Korea, 2006; In 2013, he received his Ph.
D.
degree in Engineering from the College of Engineering and Food Science of Zhejiang University.
Since 2009, he has been a teacher
at Zhejiang A&F University.
The main research directions are the intensive processing and comprehensive utilization of agricultural products, the research and development
of food nutrition and health products.
In recent years, he has presided over 5 projects of the National Natural Science Foundation of China and provincial and ministerial projects, published more than 30 papers as the first author or corresponding author, and obtained 3 national invention patents
.
Guided students to win 1 bronze award in the National Competition of the Internet + Entrepreneurship Competition and 2 provincial silver awards in the Challenge Cup Entrepreneurship Competition
.
First author bio
Deng Ke, Master candidate, College of
Food and Health, Zhejiang A&F University.
2019.
09-2021.
01, Zhejiang A&F University, Food Processing and Safety, Master of
Agronomy.
This article "Preparation of Isoflavones and Their Protective Effect on Oxidative Damage in RIN-m5F Cells" is from Food Science, Vol.
43, No.
19, 2022, pages 200-207, authors: DENG Ke, CHEN Wei, YANG Liangyuan, HU Yu, XU Guangzhi, ZHANG Youdao, WANG Yan, NI Qinxue
.
DOI:10.
7506/spkx1002-6630-20210906-067
。 Click to view information about
the article.
