Guangdong Province Research and Development Center for Chinese Medicine in Disease Susceptibility, College of Pharmacy, Jinan University, Guangzhou 510632, China; International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Chinese Ministry of Education (MOE), College of Pharmacy, Jinan University, Guangzhou 510632, China; Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, College of Pharmacy, Jinan University, Guangzhou 510632, China; Guangdong Perfect Institute for Health Science & Technology, Ltd., Zhongshan 528400, China.
Guangdong Province Research and Development Center for Chinese Medicine in Disease Susceptibility, College of Pharmacy, Jinan University, Guangzhou 510632, China; International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Chinese Ministry of Education (MOE), College of Pharmacy, Jinan University, Guangzhou 510632, China; Guangdong Province Key Laboratory of Pharmacodynamic Constituents of TCM and New Drugs Research, College of Pharmacy, Jinan University, Guangzhou 510632, China.
Phytomedicine. 2021 Jan;80:153398. doi: 10.1016/j.phymed.2020.153398. Epub 2020 Oct 24.
Celastrol, a pentacyclic triterpenoid quinonemethide isolated from several spp. of Celastraceae family, exhibits anti-inflammatory activities in a variety of diseases including arthritis.
This study aims to investigate whether the inhibition of NLRP3 inflammasome is engaged in the anti-inflammatory activities of celastrol and delineate the underlying mechanism.
The influence of celastrol on NLRP3 inflammasome activation was firstly studied in lipopolysaccharide (LPS)-primed mouse bone marrow-derived macrophages (BMDMs) and phorbol 12-myristate 13-acetate (PMA)-primed THP-1 cells treated with nigericin. Reconstituted inflammasome was also established by co-transfecting NLRP3, ASC, pro-caspase-1 and pro-IL-1β in HEK293T cells. The changes of inflammasome components including NLRP3, ASC, pro-caspase-1/caspase-1 and pro-IL-1β/IL-1β were examined by enzyme-linked immunosorbent assay (ELISA), western blotting and immunofluorescence. Furthermore, Propionibacterium acnes (P. acnes)/LPS-induced liver injury and monosodium urate (MSU)-induced gouty arthritis in mice were employed in vivo to validate the inhibitory effect of celastrol on NLRP3 inflammasome.
Celastrol significantly suppressed the cleavage of pro-caspase-1 and pro-IL-1β, while not affecting the protein expressions of NLRP3, ASC, pro-caspase-1 and pro-IL-1β in THP-1 cells, BMDMs and HEK293T cells. Celastrol suppressed NLRP3 inflammasome activation and alleviated P. acnes/LPS-induced liver damage and MSU-induced gouty arthritis. Mechanism study revealed that celastrol could interdict K63 deubiquitination of NLRP3, which may concern interaction of celastrol and BRCA1/BRCA2-containing complex subunit 3 (BRCC3), and thereby prohibited the formation of NLRP3, ASC and pro-caspase-1 complex to block the generation of mature IL-1β.
Celastrol suppresses NLRP3 inflammasome activation in P. acnes/LPS-induced liver damage and MSU-induced gouty arthritis via inhibiting K63 deubiquitination of NLRP3, which presents a novel insight into inhibition of celastrol on NLRP3 inflammasome and provides more evidences for its application in the therapy of inflammation-related diseases.
从卫矛科几种植物中分离得到的五环三萜醌甲醚化合物雷公藤红素具有抗炎活性，可用于治疗关节炎等多种疾病。
本研究旨在探讨雷公藤红素是否通过抑制 NLRP3 炎性小体参与其抗炎作用，并阐明其潜在机制。
首先在脂多糖（LPS）诱导的小鼠骨髓来源巨噬细胞（BMDMs）和佛波醇 12-肉豆蔻酸 13-乙酸酯（PMA）诱导的 THP-1 细胞中用 Nigericin 处理，研究雷公藤红素对 NLRP3 炎性小体激活的影响。重组炎性小体通过共转染 NLRP3、ASC、前胱天蛋白酶-1 和前白细胞介素-1β（pro-IL-1β）在 HEK293T 细胞中建立。通过酶联免疫吸附试验（ELISA）、western blot 和免疫荧光检测炎性小体成分 NLRP3、ASC、前胱天蛋白酶-1/胱天蛋白酶-1 和前白细胞介素-1β/白细胞介素-1β的变化。此外，在体内利用痤疮丙酸杆菌（P. acnes）/LPS 诱导的肝损伤和单钠尿酸盐（MSU）诱导的痛风性关节炎模型验证雷公藤红素对 NLRP3 炎性小体的抑制作用。
雷公藤红素显著抑制前胱天蛋白酶-1 和前白细胞介素-1β的裂解，而不影响 THP-1 细胞、BMDMs 和 HEK293T 细胞中 NLRP3、ASC、前胱天蛋白酶-1 和前白细胞介素-1β的蛋白表达。雷公藤红素抑制 NLRP3 炎性小体激活，并减轻 P. acnes/LPS 诱导的肝损伤和 MSU 诱导的痛风性关节炎。机制研究表明，雷公藤红素可以抑制 NLRP3 的 K63 去泛素化，这可能与雷公藤红素和 BRCA1/BRCA2 含复合物亚基 3（BRCC3）的相互作用有关，从而阻止 NLRP3、ASC 和前胱天蛋白酶-1 复合物的形成，阻断成熟白细胞介素-1β的产生。
雷公藤红素通过抑制 NLRP3 的 K63 去泛素化抑制 P. acnes/LPS 诱导的肝损伤和 MSU 诱导的痛风性关节炎中的 NLRP3 炎性小体激活，为雷公藤红素抑制 NLRP3 炎性小体提供了新的见解，并为其在炎症相关疾病的治疗提供了更多证据。
