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雪松醇通过促进miR-542-5p表达减轻白细胞介素-1β处理的软骨细胞的凋亡和炎症反应。

Cedrol alleviates the apoptosis and inflammatory response of IL-1β-treated chondrocytes by promoting miR-542-5p expression.

作者信息

Dong Wangchao, Wang Shanshan, Qian Weiqing, Li Suming, Wang Peimin

机构信息

Department of Orthopedics, Nanjing Hospital of Chinese Medicine, Nanjing, China.

Department of Pharmacy, Children's Hospital of Nanjing Medical University, Nanjing, China.

出版信息

In Vitro Cell Dev Biol Anim. 2021 Dec;57(10):962-972. doi: 10.1007/s11626-021-00620-3. Epub 2022 Dec 10.

Abstract

Cedrol has been shown to exert anti-tumor, anti-inflammatory, and anti-oxidative effects, but its role in osteoarthritis (OA) is unclear. This study aimed to explore the effect of cedrol in OA. Chondrocytes were isolated from newborn rats and cultured in Dulbecco's modified Eagle's medium (DMEM). Then, Alcian blue staining was used to identify the chondrocytes. IL-1β and cedrol were used to treat chondrocytes. Cell viability and apoptosis were measured by MTT and flow cytometry assays, respectively. The expressions of miR-542-5p, miR-26b-5p, miR-572, miR-138-5p, miR-328-3p, miR-1254, Bcl-2, Bax, iNOS, COX-2, and MMP-13 were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR) or western blot. NO and PGE2 levels were detected by ELISA. All the cells extracted from the newborn rats were dyed blue, indicating that the cells were chondrocytes. IL-1β could reduce the viability and promote apoptosis and inflammatory response of chondrocytes, while cedrol could reverse the effect of IL-1β. In addition, cedrol could significantly increase the expression of miR-542-5p in IL-1β-treated chondrocytes. Moreover, miR-542-5p inhibitor could partly reverse the effect of cedrol in the apoptosis and inflammation response of chondrocytes. Cedrol alleviated IL-1β-induced apoptosis and inflammatory response of chondrocytes by promoting miR-542-5p expression.

摘要

雪松醇已被证明具有抗肿瘤、抗炎和抗氧化作用,但其在骨关节炎(OA)中的作用尚不清楚。本研究旨在探讨雪松醇在骨关节炎中的作用。从新生大鼠中分离软骨细胞,并在杜尔贝科改良的 Eagle 培养基(DMEM)中培养。然后,使用阿尔辛蓝染色来鉴定软骨细胞。用白细胞介素 -1β(IL-1β)和雪松醇处理软骨细胞。分别通过 MTT 法和流式细胞术检测细胞活力和凋亡情况。通过定量逆转录聚合酶链反应(qRT-PCR)或蛋白质免疫印迹法检测 miR-542-5p、miR-26b-5p、miR-572、miR-138-5p、miR-328-3p、miR-1254、Bcl-2、Bax、诱导型一氧化氮合酶(iNOS)、环氧化酶 -2(COX-2)和基质金属蛋白酶 -13(MMP-13)的表达。通过酶联免疫吸附测定(ELISA)检测一氧化氮(NO)和前列腺素 E2(PGE2)水平。从新生大鼠中提取的所有细胞均被染成蓝色,表明这些细胞是软骨细胞。IL-1β 可降低软骨细胞的活力,促进其凋亡和炎症反应,而雪松醇可逆转 IL-1β 的作用。此外,雪松醇可显著增加经 IL-1β 处理的软骨细胞中 miR-542-5p 的表达。而且,miR-542-5p 抑制剂可部分逆转雪松醇对软骨细胞凋亡和炎症反应的影响。雪松醇通过促进 miR-542-5p 的表达减轻了 IL-1β 诱导的软骨细胞凋亡和炎症反应。

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