不同方法对鹰嘴豆总黄酮提取含量的影响及鹰嘴豆总黄酮对小鼠的镇痛效果研究

doi:10.12160/j.issn.1672-5190.2024.02.011

摘要/Abstract

摘要： [目的]研究鹰嘴豆总黄酮最佳提取及处理方法,评价鹰嘴豆总黄酮对小鼠的镇痛作用。[方法]以产自新疆维吾尔自治区的鹰嘴豆为试验材料,采用硝酸铝-亚硝酸钠比色法测定经超声波辅助提取法、索氏回流提取法、微波提取法、超声波辅助提取+旋转蒸发处理法、索氏回流提取+旋转蒸发处理法、超声波辅助提取+冷冻干燥处理法分析获得的鹰嘴豆总黄酮含量,确定鹰嘴豆总黄酮的最佳提取及处理方式。将昆明系小鼠随机分为5组,鹰嘴豆总黄酮高、中、低剂量组分别灌胃鹰嘴豆总黄酮60、30、15 mg/（kg·BW）,阳性对照组灌胃阿司匹林80 mg/（kg·BW）,空白对照组灌胃生理盐水0.1 mL/（10 g·BW）,每日给药1次,连续3 d;按照上述分组及给药方案,采用热板舔足法（n=40,每组8只）、温浴甩尾法（n=40,每组8只）和醋酸扭体法（n=40,每组8只）考查不同剂量鹰嘴豆总黄酮对小鼠的镇痛效果。[结果]不同提取及处理方法获得的鹰嘴豆总黄酮含量为：超声波辅助提取+冷冻干燥处理法（51.18 mg/g）>索氏回流提取法（13.16 mg/g）>超声波辅助提取法（4.86 mg/g）>索氏回流提取+旋转蒸发处理法（4.75 mg/g）>微波提取法（3.03 mg/g）>超声波辅助提取+旋转蒸发处理法（0.57 mg/g）。与空白对照组相比,高剂量和低剂量鹰嘴豆总黄酮分别显著（P<0.05）和极显著（P<0.01）延长给药后10、30、60 min小鼠热板舔足反应痛阈,中剂量鹰嘴豆总黄酮可显著（P<0.05）延长给药后10、30 min小鼠热板舔足反应痛阈;中剂量和低剂量鹰嘴豆总黄酮均可极显著（P<0.01）延长给药后2 h小鼠温浴甩尾反应痛阈;高、中、低剂量鹰嘴豆总黄酮均可极显著（P<0.01）减少醋酸致小鼠扭体反应次数,高、中、低剂量组的疼痛抑制率分别为35.44%、35.44%、61.37%,低剂量组的扭体反应次数低于阳性对照组,疼痛抑制率高于阳性对照组。[结论]确定的鹰嘴豆总黄酮最优提取及处理方法是超声波辅助提取+冷冻干燥处理法。不同剂量的鹰嘴豆总黄酮对小鼠均表现出较好的镇痛效果,以低剂量[15 mg/（kg·BW）]鹰嘴豆总黄酮的镇痛效果最好。

关键词: 鹰嘴豆, 总黄酮, 提取方法, 超声波辅助提取, 冷冻干燥处理, 镇痛效果

Abstract: [Objective] This study aimed to determine the optimal extraction and processing method for total flavonoids from chickpea （Cicer arietinum）, and to evaluate the analgesic effect of chickpea total flavonoids （CTFs） on mice. [Method] The chickpeas produced in Xinjiang Uygur Autonomous Region were used as the experimental materials. The aluminum nitrate-sodium nitrite colorimetric method was employed to assess the contents of CTFs obtained through ultrasonic-assisted extraction, Soxhlet reflux extraction, micro-wave extraction, ultrasonic-assisted extraction+rotary evaporation treatment, Soxhlet reflux extraction+rotary evaporation treatment, ultrasonic-assisted extraction+freeze-drying treatment, respectively, to determine the optimal extraction and processing method for CTFs. Kunming mice were randomly assigned into 5 groups. The high, medium and low dose groups were gavaged with 60, 30 and 15 mg/(kg · BW) of CTFs. The positive control group was gavaged with 80 mg/（kg·BW） of aspirin. The blank control group was gavaged with 0.1 mL/（10 g·BW） of physiological saline. Each group was administered once daily for 3 consecutive days. According to the above grouping and administration scheme, the analgesic effects of different doses of CTFs on mice were evaluated using hot plate test （n=40, 8 mice in each group）, warm bath test （n=40, 8 mice in each group） and acetic acid twisting test （n=40, 8 mice in each group）. [Result] The CTFs obtained by different extraction and processing methods were as follows: ultrasonic-assisted extraction+freeze-drying treatment （51.18 mg/g）>Soxhlet reflux extraction （13.16 mg/g）>ultrasonic-assisted extraction （4.86 mg/g）>Soxhlet reflux extraction+rotary evaporation treatment （4.75 mg/g）>micro-wave extraction （3.03 mg/g）>ultrasonic-assisted extraction+rotary evaporation treatment （0.57 mg/g）. Hot plate test demonstrated that compared with the blank control group, CTFs high and low dose groups had extremely significantly （P<0.01） prolonged pain threshold at 10, 30 and 60 min after administration, while CTFs medium dose group exhibited significantly （P<0.05） prolonged pain threshold at 10 and 30 min after administration. CTFs medium and low dose groups had extremely significantly （P<0.01） prolonged pain threshold in warm bath test at 2 h after administration than the blank control group. CTFs high, medium and low dose groups had extremely significantly （P<0.01） reduced the frequency of twisting induced by acetic acid compared with the blank control group, with the pain inhibition rates of 35.44%, 35.44% and 61.37%, respectively. In addition, the frequency of twisting in CTFs low dose group was lower than that in the positive control group, with higher pain inhibition rate than that in the positive control group. [Conclusion] The optimal extraction and processing method for CTFs was ultrasound-assisted extraction+freeze-drying treatment. Different doses of CTFs had good analgesic effects on mice, with low dose [15 mg/（kg·BW）] having the best.

Key words: chickpea, total flavonoids, extraction method, ultrasonic-assisted extraction, freeze-drying treatment, analgesic effect

中图分类号:

S529
S859.79

海婷玉, 金科旭, 阿力米热·阿布都外力, 冯浩强, 戴小华. 不同方法对鹰嘴豆总黄酮提取含量的影响及鹰嘴豆总黄酮对小鼠的镇痛效果研究[J]. 畜牧与饲料科学, 2024, 45(2): 88-94.

HAI Tingyu, JIN Kexu, ELMYRA Abduwali, FENG Haoqiang, DAI Xiaohua. Impacts of Different Procedures on the Extraction of Total Flavonoids from Chickpea（Cicer arietinum）and the Analgesic Effect on Mice[J]. Animal Husbandry and Feed Science, 2024, 45(2): 88-94.

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