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[白芍汤制黄药子减毒炮制工艺及机制研究]

[Toxicity attenuation processing technology and mechanism of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction].

作者信息

Li Bing-Yin, Wang Jun-Ming, Song Ling-Ling, Duan Ya-Qian, Long Bing-Yu, Qin Ling-Yu, Wu Xiao-Hui, Wang Yan-Mei, Gong Ming-Zhu

机构信息

College of Pharmacy, Henan University of Chinese Medicine Zhengzhou 450046, China.

College of Pharmacy, Henan University of Chinese Medicine Zhengzhou 450046, China Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan &Education Ministry, Henan University of Chinese Medicine Zhengzhou 450046, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2023 May;48(9):2455-2463. doi: 10.19540/j.cnki.cjcmm.20230117.301.

Abstract

This study explored toxicity attenuation processing technology of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction for the first time, and further explored its detoxification mechanism. Nine processed products of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction were prepared by orthogonal experiment with three factors and three levels. Based on the decrease in the content of the main hepatotoxic component diosbulbin B before and after processing of Rhizoma Dioscoreae Bulbiferae by high-performance liquid chromatography, the toxicity attenuation technology was preliminarily screened out. On this basis, the raw and representative processed products of Rhizoma Dioscoreae Bulbiferae were given to mice by gavage with 2 g·kg~(-1)(equival to clinical equivalent dose) for 21 d. The serum and liver tissues were collected after the last administration for 24 h. The serum biochemical indexes reflecting liver function and liver histopathology were combined to further screen out and verify the proces-sing technology. Then, the lipid peroxidation and antioxidant indexes of liver tissue were detected by kit method, and the expressions of NADPH quinone oxidoreductase 1(NQO1) and glutamate-cysteine ligase(GCLM) in mice liver were detected by Western blot to further explore detoxification mechanism. The results showed that the processed products of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction reduced the content of diosbulbin B and improved the liver injury induced by Rhizoma Dioscoreae Bul-biferae to varying degrees, and the processing technology of A_2B_2C_3 reduced the excessive levels of alanine transaminase(ALT) and aspartate transaminase(AST) induced by raw Rhizoma Dioscoreae Bulbiferae by 50.2% and 42.4%, respectively(P<0.01, P<0.01). The processed products of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction reversed the decrease protein expression levels of NQO1 and GCLM in the liver of mice induced by raw Rhizoma Dioscoreae Bulbiferae to varying degrees(P<0.05 or P<0.01), and it also reversed the increasing level of malondialdehyde(MDA) and the decreasing levels of glutathione(GSH), glutathione peroxidase(GPX), and glutathione S-transferase(GST) in the liver of mice(P<0.05 or P<0.01). In summary, this study shows that the optimal toxicity attenuation processing technology of Rhizoma Dioscoreae Bulbiferae stir-fried with Paeoniae Radix Alba decoction is A_2B_2C_3, that is, 10% of Paeoniae Radix Alba decoction is used for moistening Rhizoma Dioscoreae Bulbiferae and processed at 130 ℃ for 11 min. The detoxification mechanism involves enhancing the expression levels of NQO1 and GCLM antio-xidant proteins and related antioxidant enzymes in the liver.

摘要

本研究首次探索了用白芍水煎液炒制黄药子的毒性减毒加工工艺,并进一步探讨其解毒机制。采用三因素三水平正交试验制备了9种白芍水煎液炒制黄药子的炮制品。基于高效液相色谱法测定黄药子炮制前后主要肝毒性成分黄独素B含量的降低,初步筛选出毒性减毒工艺。在此基础上,将黄药子的生品和代表性炮制品按2 g·kg⁻¹(相当于临床等效剂量)灌胃给予小鼠,连续21天。末次给药24小时后采集血清和肝组织。结合反映肝功能的血清生化指标和肝脏组织病理学进一步筛选和验证加工工艺。然后,采用试剂盒法检测肝组织的脂质过氧化和抗氧化指标,采用蛋白质免疫印迹法检测小鼠肝脏中NADPH醌氧化还原酶1(NQO1)和谷氨酸-半胱氨酸连接酶(GCLM)的表达,进一步探讨解毒机制。结果表明,白芍水煎液炒制黄药子的炮制品降低了黄独素B的含量,不同程度地改善了黄药子所致的肝损伤,其中A₂B₂C₃加工工艺使生黄药子所致的谷丙转氨酶(ALT)和谷草转氨酶(AST)过高水平分别降低了50.2%和42.4%(P<0.01,P<0.01)。白芍水煎液炒制黄药子的炮制品不同程度地逆转了生黄药子所致小鼠肝脏中NQO1和GCLM蛋白表达水平的降低(P<0.05或P<0.01),也逆转了小鼠肝脏中丙二醛(MDA)水平的升高以及谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GPX)和谷胱甘肽S-转移酶(GST)水平的降低(P<0.05或P<0.0(1)。综上所述,本研究表明白芍水煎液炒制黄药子的最佳毒性减毒加工工艺为A₂B₂C₃,即采用10%的白芍水煎液润制黄药子,在130℃下炒制11分钟。解毒机制涉及增强肝脏中NQO1和GCLM抗氧化蛋白及相关抗氧化酶的表达水平。

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