West China School of Public Health and West China Fourth Hospital, Sichuan University, Renmin South Road, Chengdu, 610041, China.
Food Safety Monitoring and Risk Assessment Key Laboratory of Sichuan Province, Department of Public Health Laboratory Sciences, West China School of Public Health, Sichuan University, Renmin South Road, Chengdu, 610041, China.
Chem Biodivers. 2023 May;20(5):e202201157. doi: 10.1002/cbdv.202201157. Epub 2023 May 2.
Our previous study has exhibited that one kind of Zanthoxylum bungeanum seed oil (ZSO), extracted from Zanthoxylum bungeanum seed, had inhibitory effects on osteoclastogenesis. However, the anti-osteoclastogenesis activities of different kinds of ZSO are scarcely reported. Since inflammation is related to bone loss and osteoporosis, in this study, three kinds of ZSO, Zanthoxylum schinifolium Siebold et Zucc seed oil (ZSSO), Zanthoxylum armatum DC. seed oil (ZDSO) and Zanthoxylum bungeanum maximum seed oil (ZBSO), were obtained with Soxhlet extraction and their fatty acid constituents were detected by GC-FID. RAW264.7 macrophages induced by lipopolysaccharide (LPS) were used to evaluate the inhibitory effects of three kinds of ZSO on inflammation via detecting the expression levels of inflammatory factors by RT-qPCR. Moreover, RANKL-induced osteoclastogenesis was applied to demonstrate the anti-osteoclastogenesis activities of them through tartrate-resistant acid phosphatase (TRAP) staining and RT-qPCR. The GC-FID results exhibited that the highest constituent in ZSSO and ZDSO was oleic acid (OA) and palmitoleic acid (PLA), respectively. While linoleic acid (LA) and α-Linolenic acid (ALA) in ZBSO were dominant. At the concentration of 0.5 μL/mL, all three kinds of ZSO could decrease the expression levels of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1 beta (IL-1β) in LPS-induced macrophages. At the concentration of 0.25 μL/mL, only ZSSO could decrease the expression levels of iNOS and COX-2, which implied the inhibitory effects of ZSSO were stronger than other ZSOs. The number of RANKL-induced osteoclasts and the expressions of nuclear factor kappa-B (NF-κB), TNF-α and IL-6 in the cells were decreased after being treated with ZSOs at the concentration of 0.5 μL/mL, while the number of RANKL-induced osteoclasts after treated with ZBSO were less than those treated with other ZSOs, this indicated that the anti-osteoclastogenesis effect of ZBSO were stronger than other ZSOs. In conclusion, the fatty acid compositions of three major kinds of ZSO were compared and the content of unsaturated fatty acids especially ω-3 polyunsaturated fatty acids in ZBSO were the highest among them. All ZSOs tested had anti-inflammatory and anti-osteoclastogenesis activities. And their anti-osteoclastogenesis effects might be related to the suppression of the NF-κB pathway.
我们之前的研究表明，从花椒种子中提取的一种花椒籽油（ZSO）对破骨细胞生成具有抑制作用。然而，不同种类的 ZSO 的抗破骨细胞生成活性却鲜有报道。由于炎症与骨丢失和骨质疏松症有关，在本研究中，我们采用索氏提取法获得了三种花椒籽油，即花椒籽油（ZSSO）、花椒籽油（ZDSO）和花椒籽油（ZBSO），并通过 GC-FID 检测其脂肪酸成分。利用脂多糖（LPS）诱导的 RAW264.7 巨噬细胞，通过 RT-qPCR 检测炎症因子的表达水平，评估三种 ZSO 对炎症的抑制作用。此外，通过抗酒石酸酸性磷酸酶（TRAP）染色和 RT-qPCR ，研究 RANKL 诱导的破骨细胞生成，以证明它们的抗破骨细胞生成活性。GC-FID 结果表明，ZSSO 和 ZDSO 中含量最高的成分分别是油酸（OA）和棕榈油酸（PLA），而 ZBSO 中则以亚油酸（LA）和α-亚麻酸（ALA）为主。在 0.5 μL/mL 的浓度下，三种 ZSO 均可降低 LPS 诱导的巨噬细胞中诱导型一氧化氮合酶（iNOS）、环氧化酶-2（COX-2）、肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）和白细胞介素-1β（IL-1β）的表达水平。在 0.25 μL/mL 的浓度下，只有 ZSSO 可以降低 iNOS 和 COX-2 的表达水平，这表明 ZSSO 的抑制作用强于其他 ZSO。用 ZSO 处理浓度为 0.5 μL/mL 时，RANKL 诱导的破骨细胞数量和细胞中核因子 kappa-B（NF-κB）、TNF-α和 IL-6 的表达均降低，而用 ZBSO 处理时，RANKL 诱导的破骨细胞数量少于用其他 ZSO 处理时，这表明 ZBSO 的抗破骨细胞生成作用强于其他 ZSO。综上所述，对三种主要的 ZSO 进行了脂肪酸组成比较，其中 ZBSO 中的不饱和脂肪酸，尤其是 ω-3 多不饱和脂肪酸含量最高。三种 ZSO 均具有抗炎和抗破骨细胞生成活性。其抗破骨细胞生成作用可能与抑制 NF-κB 通路有关。
