Picroside II   中文名称: 胡黄连苷 II

- Target of Picroside II in sepsis model: Nuclear factor-kappa B (NF-κB) signaling pathway- Target of Picroside II in cerebral ischemia-reperfusion model: Oxidative stress-related pathways (Nrf2/HO-1, MAPK) and tight junction (TJ) protein-related signaling[2]

1. RAW264.7巨噬细胞的抗炎活性（脓毒症相关）：用脂多糖（LPS，1 μg/mL）和Picroside II（5 μM、10 μM、20 μM）处理RAW264.7细胞24小时。ELISA结果显示，Picroside II浓度依赖性降低LPS诱导的肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）分泌，20 μM时可使TNF-α和IL-6水平分别降低约65%和60%。Western blot显示，Picroside II抑制LPS诱导的NF-κB p65和IκBα磷酸化，减少p65核转位[1]
2. 脑微血管内皮细胞（BMECs）的保护作用（脑缺血相关）：对BMECs进行氧糖剥夺/复氧（OGD/R）处理，并加入Picroside II（2.5 μM、5 μM、10 μM）。跨内皮电阻（TEER）检测显示，Picroside II逆转OGD/R诱导的TEER下降，10 μM时可使TEER恢复至正常水平的约85%。DCFH-DA染色显示，Picroside II减少OGD/R诱导的活性氧（ROS）生成，10 μM时ROS水平降低约55%。Western blot显示，Picroside II上调OGD/R下调的紧密连接蛋白（ZO-1、occludin）和Nrf2/HO-1通路蛋白，同时下调磷酸化p38 MAPK[2]

1. 小鼠脓毒症模型（CLP诱导）的保护作用：对雄性C57BL/6小鼠进行盲肠结扎穿孔（CLP）构建脓毒症模型，分为假手术组、CLP模型组、Picroside II 20 mg/kg组、Picroside II 40 mg/kg组。Picroside II于CLP前1小时腹腔注射，之后每日1次，连续3天。生存率分析显示，Picroside II提高7天生存率（40 mg/kg组生存率65%，模型组20%）。组织病理学显示，Picroside II减轻CLP诱导的肺、肝、肾损伤（减少炎症细胞浸润和组织坏死）。ELISA显示，Picroside II降低血清TNF-α、IL-6、IL-1β水平（40 mg/kg使TNF-α降低约70%），Western blot显示肺组织中NF-κB激活减弱[1]
2. 大鼠脑缺血再灌注模型（MCAO诱导）的保护作用：对雄性Sprague-Dawley大鼠进行大脑中动脉阻塞（MCAO）2小时后再灌注24小时，分为假手术组、MCAO模型组、Picroside II 10 mg/kg组、Picroside II 20 mg/kg组。Picroside II于再灌注即刻尾静脉注射。神经功能缺损评分显示，Picroside II降低评分（20 mg/kg组评分1.5，模型组3.2）。TTC染色显示，Picroside II减少脑梗死体积（20 mg/kg使梗死体积降低约45%）。伊文思蓝（EB）渗漏实验显示，Picroside II降低血脑屏障通透性（20 mg/kg使EB渗漏减少约50%）。生化分析显示，Picroside II提高脑组织中超氧化物歧化酶（SOD）活性，降低丙二醛（MDA）含量[2]

1. RAW264.7巨噬细胞实验（脓毒症相关）：将RAW264.7细胞以5×10^5个/孔接种于6孔板，用含10%胎牛血清的DMEM培养。细胞融合至80%后，分为对照组、LPS组（1 μg/mL）、Picroside II组（5 μM、10 μM、20 μM + 1 μg/mL LPS）。在37°C、5% CO2条件下培养24小时后，收集细胞上清液用于ELISA（检测TNF-α、IL-6），裂解细胞用于Western blot（检测p-p65、p-IκBα、核p65）。提取核蛋白时，用核蛋白提取试剂盒处理细胞，取核组分进行Western blot[1]
2. BMEC细胞实验（脑缺血相关）：将BMECs接种于Transwell小室（用于TEER检测）或6孔板（5×10^5个/孔），用内皮细胞培养基培养。细胞形成融合单层后，用无糖DMEM替换培养基并置于1% O2环境中培养4小时诱导OGD，再换用正常培养基置于5% CO2环境中培养24小时进行复氧。复氧开始时加入Picroside II（2.5 μM、5 μM、10 μM），在复氧0、6、12、24小时检测TEER。检测ROS时，用10 μM DCFH-DA负载细胞30分钟，通过流式细胞仪检测荧光强度。Western blot检测时，裂解细胞后检测ZO-1、occludin、Nrf2、HO-1、p-p38[2]

1. Mouse sepsis model (CLP) protocol: Male C57BL/6 mice (20-25 g) were anesthetized with isoflurane. A midline abdominal incision was made, cecum was ligated (70% of cecum), and punctured twice with a 22-gauge needle. Cecum was returned to abdomen, and incision was closed. Sham group only had abdominal incision without ligation/puncture. Mice were divided into 4 groups (n=15/group): sham, CLP model, Picroside II 20 mg/kg, Picroside II 40 mg/kg. Picroside II was dissolved in normal saline (0.9% NaCl) and administered via intraperitoneal injection (0.1 mL/10 g body weight) 1 hour before CLP, then once daily on days 1-3 after CLP. On day 4, mice were sacrificed; serum was collected for ELISA, and lung, liver, kidney tissues were collected for histopathology and Western blot. Survival rate was recorded for 7 days [1]
2. Rat cerebral ischemia-reperfusion model (MCAO) protocol: Male Sprague-Dawley rats (250-300 g) were anesthetized with chloral hydrate. MCAO was performed by inserting a nylon suture into the internal carotid artery to block the middle cerebral artery for 2 hours; suture was removed for 24-hour reperfusion. Sham group only had artery dissection without suture insertion. Rats were divided into 4 groups (n=8/group): sham, MCAO model, Picroside II 10 mg/kg, Picroside II 20 mg/kg. Picroside II was dissolved in normal saline and administered via tail vein injection (0.1 mL/10 g body weight) immediately after reperfusion. After 24-hour reperfusion, neurological deficit score was evaluated (0-5 scale). Rats were then sacrificed; brain was collected for TTC staining (infarction volume) and EB leakage assay (blood-brain barrier permeability). Brain tissue homogenate was prepared to detect SOD activity and MDA content [2]

1. In mouse sepsis model: After 4-day treatment with Picroside II (20-40 mg/kg, intraperitoneal injection), serum levels of alanine transaminase (ALT), aspartate transaminase (AST), blood urea nitrogen (BUN), and creatinine (Cr) were measured. No significant differences were found between Picroside II-treated groups and sham group, indicating no hepatotoxicity or nephrotoxicity. Body weight of Picroside II-treated mice was stable, with no obvious weight loss [1]
2. In rat cerebral ischemia model: After 24-hour treatment with Picroside II (10-20 mg/kg, tail vein injection), serum ALT, AST, BUN, Cr levels were within normal ranges, and no abnormal changes in organ weight (brain, liver, kidney) were observed, suggesting no acute toxicity [2]

[1]. Picroside II protects against sepsis via suppressing inflammation in mice. Am J Transl Res. 2016 Dec 15;8(12):5519-5531. eCollection 2016.

[2]. Picroside II protects the blood-brain barrier by inhibiting the oxidative signaling pathway in cerebral ischemia-reperfusion injury. PLoS One. 2017 Apr 7;12(4):e0174414.

Picroside II has been reported in Veronica anagallis, Catalpa ovata, and other organisms with data available.

---

1. Picroside II is a major active component isolated from the roots of Picrorhiza scrophulariiflora , a traditional Chinese herb used for anti-inflammatory and hepatoprotective purposes [1][2]
2. Mechanism in sepsis: Picroside II exerts anti-sepsis effects by inhibiting the NF-κB signaling pathway, reducing the production of pro-inflammatory cytokines, and alleviating organ damage caused by excessive inflammation [1]
3. Mechanism in cerebral ischemia: Picroside II protects the blood-brain barrier by inhibiting oxidative stress (activating Nrf2/HO-1 pathway, reducing ROS/MDA) and preserving TJ protein expression, while inhibiting p38 MAPK-mediated cell damage [2]

C23H28O13

512.4606

512.153

39012-20-9

11944602

White to yellow solid powder

1.7±0.1 g/cm3

780.8±60.0 °C at 760 mmHg

267.9±26.4 °C

0.0±2.8 mmHg at 25°C

1.681

-2.5

197.13

6

13

8

36

839

11

COC1=C(C=CC(=C1)C(=O)O[C@H]2[C@@H]3C=CO[C@H]([C@@H]3[C@@]4([C@H]2O4)CO)O[C@H]5[C@@H]([C@H]([C@@H]([C@H](O5)CO)O)O)O)O

AKNILCMFRRDTEY-NUGKWEEESA-N

InChI=1S/C23H28O13/c1-31-12-6-9(2-3-11(12)26)20(30)34-18-10-4-5-32-21(14(10)23(8-25)19(18)36-23)35-22-17(29)16(28)15(27)13(7-24)33-22/h2-6,10,13-19,21-22,24-29H,7-8H2,1H3/t10-,13-,14-,15-,16+,17-,18+,19+,21+,22+,23-/m1/s1

[(1S,2S,4S,5S,6R,10S)-2-(hydroxymethyl)-10-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3,9-dioxatricyclo[4.4.0.02,4]dec-7-en-5-yl] 4-hydroxy-3-methoxybenzoate

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 : ~100 mg/mL (~195.14 mM)

配方 1 中的溶解度: ≥ 2.5 mg/mL (4.88 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% 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储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

---

配方 2 中的溶解度: ≥ 2.5 mg/mL (4.88 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。

---

View More

配方 3 中的溶解度: ≥ 2.5 mg/mL (4.88 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (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 储备液加入到 900 μL 玉米油中并混合均匀。

---

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：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。