产品介绍：
含铁血黄素(Hemosiderin)是一种血红蛋白源性色素，为金黄色或棕黄色颗粒，因其含铁，且为金黄色，故称为含铁血黄素。当红细胞被巨噬细胞吞噬后，在溶酶体酶的作用下，血红蛋白被分解为不含铁的橙色血质和含铁的含铁血黄素。Perls普鲁士蓝反应(Prussian blue reaction)又称为含铁血黄素染色，即经过亚铁氰化钾和稀酸处理后可以产生蓝色，常见于吞噬细胞或间质内，其染色原理在于亚铁氰化钾溶液使三价铁离子从蛋白质中被稀盐酸分离出来，三价铁与亚铁氰化钾反应，生成一种不溶解的蓝色化合物即三价铁的亚铁氰化物。
普鲁士蓝染色用于显示局部组织内的各种出血性病变，常见于吞噬细胞内，可以很好地区分含铁血黄素与其他色素。该染色液稳定性好、可以长期保存、不易产生沉淀、应用范围广，可以进行复染，该染色液的复染液采用中性红。
自备材料：
1、 固定液：10%中性福尔马林、4%多聚甲醛等
2、 系列乙醇
使用方法：
（一）石蜡切片染色：
1、 组织固定于10%中性福尔马林，常规脱水包埋。
2、 切片厚度4μm，常规脱蜡至水。
3、 蒸馏水水洗1min。
4、 切片入perls stain（见注意事项2），浸染15~30min。
5、 蒸馏水充分冲洗5~10min。
6、 入伊红染色液，淡染细胞核5~10min。
7、 自来水冲洗1~5s。
8、 常规脱水透明，中性树胶封固。
（二）冰冻切片染色：
1、 无需脱蜡，直接迅速骨蒸馏水冲洗2~3min。
2、 染色、脱蜡、透明、封固步骤同石蜡切片的染色步骤。
（三）细胞染色：
1、 4%多聚甲醛固定10~20min。
2、 自来水冲洗2次，每次2min。
3、 蒸馏水冲洗2次，每次2min。
4、 染色、脱蜡、透明、封固步骤同石蜡切片的染色步骤。
染色结果
含铁血黄素或三价铁 | 蓝色 |
细胞核、其他组织 | 红色 |
阴性对照（可选）
取相同切片脱蜡至水。入5%草酸孵育2~6h，经perls stain，其余步骤同上。结果为阴性。
注意事项
1、 切片脱蜡应尽量干净。组织固定常采用10%中性福尔马林，经普通福尔马林长期固定后，组织会有损伤。避免使用酸性固定剂，铬酸盐处理也会妨碍铁的保存。
2、 整个操作过程中容器要干净，避免用金属铁制品，洗切片和容器时以蒸馏水为宜，因普通水内含铁质。perls stain染色时，应根据样本情况调整着色时间。
3、 所有切片都应使用同一个阳性对照切片，选择适合的对照非常重要。尸检肺组织是一个很好的对照，包含相当数量的铁阳性巨噬细胞（心衰细胞）。
4、 冰冻切片和细胞染色，最好根据具体情况摸索实验条件。
5、 为了您的安全和健康，请穿好实验服并佩戴一次性手套和口罩操作。
6、 本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。
产品组成：
名称 | AWI0484a 2×50ml | AWI0484b 2×100ml | |
试剂（A）：perls stain | A1：perls stain A | 25ml | 50ml |
A2：perls stain B | 25ml | 50ml | |
临用前，取A1、A2等量混合即为 perls stain，不宜提前配制。 | |||
试剂（B）：中性红染色液 | 50ml | 100ml | |
注意:
1.本产品仅供科研使用。请勿用于医药、临床诊断或治疗。食品及化妆品等用途。请勿存放于普通住宅区。
2.为了您的安全和健康，请穿好实验服并佩戴一次性手套和口罩操作。
3.实验结果可由多种因素影响，相关处理只限于产品本身，不涉及其他赔偿。
参考文献 (2)
Objective Smoking was a major risk factor for chronic obstructive pulmonary disease (COPD). This study plan to explore the mechanism of Polyphyllin B in lung injury induced by cigarette smoke (CSE) in COPD. Methods Network pharmacology and molecular docking were applied to analyze the potential binding targets for Polyphyllin B and COPD. Commercial unfiltered CSE and LPS were used to construct BEAS-2B cell injury in vitro and COPD mouse models in vivo, respectively, which were treated with Polyphyllin B or fecal microbiota transplantation (FMT). CCK8, LDH and calcein-AM were used to detect the cell proliferation, LDH level and labile iron pool. Lung histopathology, Fe 3+ deposition and mitochondrial morphology were observed by hematoxylin–eosin, Prussian blue staining and transmission electron microscope, respectively. ELISA was used to measure inflammation and oxidative stress levels in cells and lung tissues. Immunohistochemistry and immunofluorescence were applied to analyze the 4-HNE, LC3 and Ferritin expression. RT-qPCR was used to detect the expression of FcRn, pIgR, STAT3 and NCOA4. Western blot was used to detect the expression of Ferritin, p-STAT3/STAT3, NCOA4, GPX4, TLR2, TLR4 and P65 proteins. 16S rRNA gene sequencing was applied to detect the gut microbiota. Results Polyphyllin B had a good binding affinity with STAT3 protein, which as a target gene in COPD. Polyphyllin B inhibited CS-induced oxidative stress, inflammation, mitochondrial damage, and ferritinophagy in COPD mice. 16S rRNA sequencing and FMT confirmed that Akkermansia and Escherichia_Shigella might be the potential microbiota for Polyphyllin B and FMT to improve CSE and LPS-induced COPD, which were exhausted by the antibiotics in C + L and C + L + P mice. CSE and LPS induced the decrease of cell viability and the ferritin and LC3 expression, and the increase of NCOA4 and p-STAT3 expression in BEAS-2B cells, which were inhibited by Polyphyllin B. Polyphyllin B promoted ferritin and LC3II/I expression, and inhibited p-STAT3 and NCOA4 expression in CSE + LPS-induced BEAS-2B cells. Conclusion Polyphyllin B improved gut microbiota disorder and inhibited STAT3/NCOA4 pathway to ameliorate lung tissue injury in CSE and LPS-induced mice.
Resveratrol is involved in regulating ferroptosis , but its role in Endometriosis (EMS) is not clear. In this study, we aim to investigate the effect of ferroptosis and resveratrol intervention in the pathogenesis of EMS cyst. Cell proliferation , migration, and oxidative stress level were analyzed. The interaction of miR-21-3p and p53 was analyzed by dual luciferase assay. The interaction between p53 and SLC7A11 were analyzed by chromatin immunoprecipitation (CHIP). The miR-21-3p, GPX4, ACSL4 , FTH1, p53, SLC7A11, Ptgs2 and Chac1 expression were analyzed by RT-qPCR or Western blot . The Fe 3+ deposition and miR-21-3p, GPX4, FTH1 and SLC7A11 expressions were increased, and ACSL4, p53, Ptgs2 and Chac1 expression were decreased in EMS patients. Resveratrol inhibited migration, induced Ptgs2 and Chac1 expression in EESCs . Overexpression of miR-21-3p inhibited p53, Ptgs2 and Chac1 expression, and promoted SLC7A11 expression, which was reversed by resveratrol. miR-21-3p bound to p53, which interacted with SLC7A11. Resveratrol promoted Ptgs2 and Chac1 expression in the sh-p53 EESCs. Resveratrol reduced miR-21-3p and SLC7A11 expressions, and increased p53, Ptgs2 and Chac1 expressions, and Fe 3+ deposition in the lesion tissues of EMS mice, which were reversed by miR-21-3p mimics. Resveratrol activated p53/SLC7A11 pathway by down-regulating miR-21-3p to promote ferroptosis and prevent the development of EMS.
