Demethyleneberberine targets AMP-activated protein kinase (AMPK) [3]
Demethyleneberberine targets nuclear factor kappa B (NF-κB) signaling pathway [2]
Demethyleneberberine acts as a mitochondria-targeted antioxidant [1]

#### 来自文献[1]
- 在乙醇处理的HepG2细胞中，去亚甲基小檗碱（Demethyleneberberine）（10、20、40 μM）以剂量依赖性方式减少细胞内活性氧（ROS）生成和线粒体超氧化物水平 [1]
- 改善线粒体膜电位（ΔΨm），增加ATP生成，缓解乙醇诱导的线粒体功能障碍 [1]
- 下调脂生成基因（SREBP-1c、FASN）表达，上调脂肪酸氧化相关基因（PPARα）表达 [1]
#### 来自文献[2]
- 在LPS诱导的RAW264.7巨噬细胞中，去亚甲基小檗碱（Demethyleneberberine）（5、10、20 μM）抑制促炎细胞因子（TNF-α、IL-6、IL-1β）和一氧化氮（NO）的产生 [2]
- 通过抑制IκBα磷酸化和p65核转位，抑制NF-κB激活 [2]
- 在TNF-α处理的Caco-2肠上皮细胞中，该化合物增强紧密连接蛋白（ZO-1、occludin）表达，改善肠屏障完整性 [2]
#### 来自文献[3]
- 在棕榈酸（PA）处理的HepG2细胞中，去亚甲基小檗碱（Demethyleneberberine）（10、20、40 μM）减少脂滴积累（油红O染色）和细胞内甘油三酯（TG）含量 [3]
- 激活AMPK磷酸化（p-AMPK），上调下游能量代谢相关基因（PGC-1α、CPT1A）[3]
- 降低PA诱导的ROS生成和丙二醛（MDA）水平，同时提高超氧化物歧化酶（SOD）和谷胱甘肽（GSH）活性 [3]

#### 来自文献[1]（酒精性肝病模型）
- 给予C57BL/6小鼠含乙醇的Lieber-DeCarli饮食4周，口服去亚甲基小檗碱（Demethyleneberberine）（25、50 mg/kg/天）以剂量依赖性方式减少肝脂肪变性（H&E染色）和肝TG含量 [1]
- 改善肝组织线粒体功能，表现为线粒体DNA拷贝数增加及复合体I/III/IV活性升高 [1]
- 减轻肝脏氧化应激（MDA降低，SOD/GSH-Px活性升高），抑制肝细胞凋亡（TUNEL染色）[1]
#### 来自文献[2]（炎症性肠病模型）
- 在DSS诱导的结肠炎小鼠中，口服去亚甲基小檗碱（Demethyleneberberine）（25、50 mg/kg/天）持续7天，缓解体重下降、结肠缩短和组织学损伤（H&E染色）[2]
- 降低结肠促炎细胞因子（TNF-α、IL-6、IL-1β）水平，抑制NF-κB p65核转位 [2]
- 通过增加肠系膜淋巴结中Treg细胞比例、降低Th1/Th17细胞比例，恢复T辅助细胞稳态 [2]
#### 来自文献[3]（非酒精性脂肪肝模型）
- 在ob/ob小鼠或高脂饮食（HFD）喂养的C57BL/6小鼠中，口服去亚甲基小檗碱（Demethyleneberberine）（25、50 mg/kg/天）持续8周，减少肝脂肪变性和肝重量 [3]
- 激活肝脏AMPK磷酸化，上调脂肪酸氧化相关蛋白（CPT1A、PGC-1α）[3]
- 降低肝脏氧化应激和炎症反应（TNF-α、IL-6 mRNA表达降低）[3]

#### 来自文献[1]（线粒体功能检测）
- 线粒体膜电位（ΔΨm）检测：HepG2细胞负载ΔΨm敏感荧光染料，经去亚甲基小檗碱（Demethyleneberberine）（10、20、40 μM）和乙醇处理后，流式细胞仪检测荧光强度 [1]
- ATP检测：乙醇处理的HepG2细胞裂解后，采用荧光素酶法试剂盒定量ATP含量，结果按蛋白浓度归一化 [1]
- 氧化应激检测：细胞/组织匀浆后，采用比色法试剂盒检测SOD、GSH-Px活性及MDA水平 [1]
#### 来自文献[3]（AMPK活性检测）
- HepG2细胞经去亚甲基小檗碱（Demethyleneberberine）（10、20、40 μM）和棕榈酸处理后裂解 [3]
- 通过Western blot检测AMPK底物（ACC）的磷酸化水平，以p-ACC/ACC比值反映AMPK激活程度 [3]

#### 来自文献[1]
- HepG2细胞在含10%胎牛血清的DMEM培养基中培养，经去亚甲基小檗碱（Demethyleneberberine）（10、20、40 μM）预处理2小时后，用100 mM乙醇处理24小时 [1]
- ROS检测：细胞负载DCFH-DA荧光探针，孵育30分钟后，流式细胞仪检测荧光强度 [1]
- 脂积累检测：细胞经油红O染色后，异丙醇洗脱，510 nm处测定吸光度 [1]
- Western blot：细胞裂解后，蛋白经SDS-PAGE分离，用SREBP-1c、FASN、PPARα及β-肌动蛋白抗体进行印迹 [1]
#### 来自文献[2]
- RAW264.7细胞经去亚甲基小檗碱（Demethyleneberberine）（5、10、20 μM）预处理1小时后，用1 μg/mL LPS刺激24小时 [2]
- 细胞因子检测：收集培养上清液，ELISA法测定TNF-α、IL-6、IL-1β水平 [2]
- NF-κB转位检测：细胞固定、透化后，用p65抗体和DAPI染色，激光共聚焦显微镜观察 [2]
- Caco-2细胞经去亚甲基小檗碱（Demethyleneberberine）（5、10、20 μM）和10 ng/mL TNF-α处理24小时；Western blot检测紧密连接蛋白 [2]
#### 来自文献[3]
- HepG2细胞经去亚甲基小檗碱（Demethyleneberberine）（10、20、40 μM）和200 μM棕榈酸处理24小时 [3]
- 甘油三酯检测：细胞裂解后，比色法测定TG含量 [3]
- Western blot：用p-AMPK、AMPK、PGC-1α、CPT1A及β-肌动蛋白抗体进行印迹 [3]

#### From Literature [1] (Alcoholic Liver Disease)
- Male C57BL/6 mice (6–8 weeks old) were randomly divided into control, ethanol model, and Demethyleneberberine (25, 50 mg/kg/day) groups (n=8 per group) [1]
- Mice were fed a Lieber-DeCarli ethanol diet (5% ethanol) for 4 weeks; the compound was dissolved in 0.5% carboxymethylcellulose sodium and administered by oral gavage once daily [1]
- At the end of treatment, mice were sacrificed; liver tissues were collected for H&E staining, TG assay, mitochondrial function analysis, and Western blot [1]
- Serum was collected to detect ALT, AST levels [1]
#### From Literature [2] (Inflammatory Bowel Disease)
- Male C57BL/6 mice (6–8 weeks old) were randomly divided into control, DSS model, and Demethyleneberberine (25, 50 mg/kg/day) groups (n=8 per group) [2]
- Colitis was induced by adding 3% DSS to drinking water for 7 days; the compound was administered by oral gavage once daily during DSS exposure [2]
- Body weight and colon length were measured; colon tissues were collected for H&E staining, cytokine assay, and Western blot [2]
- Mesenteric lymph nodes were isolated for flow cytometry analysis of T-cell subsets [2]
#### From Literature [3] (Non-Alcoholic Fatty Liver Disease)
- Male ob/ob mice or HFD-fed C57BL/6 mice (6–8 weeks old) were randomly divided into control, model, and Demethyleneberberine (25, 50 mg/kg/day) groups (n=8 per group) [3]
- The compound was dissolved in 0.5% carboxymethylcellulose sodium and administered by oral gavage once daily for 8 weeks [3]
- Mice were sacrificed; liver tissues were collected for H&E staining, TG assay, and Western blot [3]
- Serum was collected to detect glucose, insulin, and lipid profiles [3]

In all animal models, oral administration of methylene berberine (25–50 mg/kg/day) for 4–8 weeks did not cause significant changes in body weight, liver function (ALT, AST), or kidney function (BUN, Cr) [1][2][3]. No obvious clinical toxicities (somnolence, loss of appetite, organ damage) were observed in the treatment group mice [1][2][3].

[1]. Demethyleneberberine, a natural mitochondria-targeted antioxidant, inhibits mitochondrial dysfunction, oxidative stress, and steatosis in alcoholic liver disease mouse model. J Pharmacol Exp Ther. 2015 Jan;352(1):139-47.

[2]. Demethyleneberberine alleviates inflammatory bowel disease in mice through regulating NF-κB signaling and T-helper cell homeostasis. Inflamm Res. 2017 Feb;66(2):187-196.

[3]. Demethyleneberberine attenuates non-alcoholic fatty liver disease with activation of AMPK and inhibition of oxidative stress. Biochem Biophys Res Commun. 2016 Apr 15;472(4):603-9.

Demethylberberine belongs to the isoquinoline class of compounds. It has been reported that demethylberberine is found in Thalictrum javanicum, and related data have been reported. Demethylberberine is a natural isoquinoline alkaloid isolated from plants of the Berberidaceae family [1][2][3]. Its core mechanisms include mitochondrial-targeted antioxidant activity, AMPK activation, and NF-κB signaling pathway inhibition [1][2][3]. It has therapeutic potential for metabolic and inflammatory diseases, including alcoholic liver disease, non-alcoholic fatty liver disease, and inflammatory bowel disease [1][2][3]. This compound exerts tissue protective effects by regulating lipid metabolism, reducing oxidative stress, inhibiting inflammation, and maintaining cell barrier function [1][2][3].

C19H18NO4

324.3505

323.116

25459-91-0

Demethyleneberberine chloride;16705-03-6

363209

Light yellow to yellow solid powder

1.649

59.64

2

4

2

24

447

0

HVTCKKMWZDDWOY-UHFFFAOYSA-O

InChI=1S/C19H17NO4/c1-23-18-4-3-11-7-15-13-9-17(22)16(21)8-12(13)5-6-20(15)10-14(11)19(18)24-2/h3-4,7-10,22H,5-6H2,1-2H3/p+1

9,10-dimethoxy-5,6-dihydroisoquinolino[2,1-b]isoquinolin-7-ium-2,3-diol

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 本产品在运输和储存过程中需避光。

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~10 mg/mL (~30.83 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (7.71 mM) (饱和度未知) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
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