责任编辑：食品科学
国家食用菌中心依托精准营养创新引智基地、河南省国际联合实验室平台，联合埃及国家农业中心Adel研究员对N. sativa进行了持续的研究，该成果以“The immunomodulatory effects of active ingredients from Nigella sativa in RAW264.7 cells through NF-κB/MAPK signaling pathways.”为题发表于中科院TOP 1区《Frontier in Nutrition》杂志（IF：6.576）。毛茛科黑种草属植物Nigella sativa原产于西南亚，主要分布在欧洲南部、亚洲西南部和北非等地区。N. sativa干燥种子具有多种药理活性，包括抗癌、菌、降压、保肝、抗炎、解热等，医疗上常用作膳食营养补充剂、祛风剂、利尿剂和驱虫剂。
Nigella sativa
Nigella sativa种子
N. sativa种子最有价值的特性之一。 近20年来，
N. sativa种子及其主要活性成分百里醌（TQ）对免疫系统的有益作用的研究取得了显著进展。例如，TQ提高T细胞中CD8和CD4的比值，改善NK细胞的功能发挥免疫调节特性。但是，对
N. sativa种子中单体化合物的免疫作用研究较少。对从
N. sativa种子70%乙醇提取物分离鉴定得到的30多个化合物活性筛选时发现，两个单糖衍生物，Ethyl-α-D-galactopyranoside （EG）和Methyl-α-D-glucoside（MG）以及两个三萜类化合物，3-
O-[
β–
D-xylopyranose-(1→3)-
α–
L-rhamnose-(1→2)-
α–
L-arabinose]-28-
O-[
α–
L-rhamnose-(1→4)-
β–
D-glucopyranose-
L-(1→6)-
β–
D-glucopyranose]-hederagenin（HXRARG）和3-
O-[
β–
D-xylopyranose-(1→3)-
α–
L-rhamnose-(1→2)-
α–
L-arabinose]-hederagenin （HXRA）能够促进RAW264.7巨噬细胞释放NO。因此，本研究以EG、MG、HXRARG和HXRA为对象，探讨
N. sativa种子的免疫增强作用和相关机制。
Fig. 1 Structures of compounds EG, MG, HXRARG and HXRA from Nigella sativa
Results and Discussio
的检测
Fig. 2 Effects of EG (A), MG (B), HXRARG (C) and HXRA (D) on nitric oxide (NO) production in RAW264.7 cells. Data are expressed as the means ± SD. Compared with the control, **P<0.01, ***P<0.001.
(A-E): microscope picture,
(F-I): quantitative analysis picture. Data are expressed as the means ± SD. Compared with the control,
**
P<0.01,
***P<0.001.
，
COX-2
蛋白表达和基因转录
(A-D) and mRNA
(E-F) expression in RAW264.7 cells.
(A-D): the left side was the protein band diagram, and the right side was the protein quantification diagram.
(E)
: mRNA expression of iNOS.
(F): mRNA expression ofCOX-2. Data are expressed as the means ± SD. Compared with the control,
**
P<0.01,
***P<0.001.
α
，
IL-6
的生成及其
mRNA
表达
α、IL-6 mRNA转录而刺激RAW264.7分泌更多的TNF-
α和IL-6。
Fig. 5 Effects of EG, MG, HXRARG and HXRA on the expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and mRNA in RAW264.7 cells. (A): secretion of IL-6 (B): secretion of TNF-α (C): TNF-α mRNA transcription (D): IL-6 mRNA transcription. Data are expressed as the means ± SD. Data are expressed as the means ±SD. Compared with the control, ***P<0.001.
ROS
的影响
(A), MG
(B), HXRARG
(C) and HXRA
(D) on ROS production ability of RAW 264.7 cells.
NF-
κ
B
信号通路的影响
κB是在免疫过程中普遍存在的一类重要的核转录因子，在许多基因的启动子和增强子中都存在功能性的NF-
κB结合位点，主要影响机体免疫有关的基因表达，是介导巨噬细胞成熟的一个关键通路。
κB P65和I
κB-
α的蛋白表达无明显影响，但I
κB-
α和P65的磷酸化蛋白表达增加，p-P65/P65，p-I
κB-
α/I
κB-
α的比值显著增大。以上结果说明，EG、MG、Hxrarg和Hxra可以激活RAW264.7细胞中的NF-
κB信号通路，从而诱导RAW264.7细胞的活化。
(A), MG
(B), HXRARG
(C) and HXRA
(D) on the expression of related proteins in NF-κB signaling pathway in RAW264.7 cells. The left side was the protein band diagram, and the right side was the protein quantification diagram. Data are expressed as the means ± SD. Compared with the control,
***P<0.001.
MAPK
信号通路的影响
MAPK家族包含三个主要的激酶，即Erk1/2，JNK和P38 MAPK。 这些激酶的共同激活促进细胞增殖、迁移、侵袭、血管生成、转移和凋亡，这对巨噬细胞和T细胞的发育和活化非常重要。
EG、MG、HXRARG或HXRA刺激RAW264.7细胞后，p-P38/P38、p-Erk/Erk或p-JNK/JNK比值均显著高于空白对照组。 提示EG、MG、HXRARG或HXRA可激活MAPK信号通路，诱导免疫应答。
(A), MG
(B), HXRARG
(C) and HXRA
(D) on the expression of related proteins in MAPK signaling pathway in RAW264.7 cells. The left side was the protein band diagram, and the right side was the protein quantification diagram. Data are expressed as the means ± SD. Compared with the control,
***P<0.001.
N. sativa种子中分离得到的四个化合物Ethyl-α-D-galactopyranoside （EG）、Methyl-α-D-glucoside（MG）、3-
O-[
β–
D-xylopyranose-(1→3)-
α–
L-rhamnose-(1→2)-
α–
L-arabinose]-28-
O-[
α–
L-rhamnose-(1→4)-
β–
D-glucopyranose-
L-(1→6)-
β–
D-glucopyranose]-hederagenin（HXRARG）和3-
O-[
β–
D-xylopyranose-(1→3)-
α–
L-rhamnose-(1→2)-
α–
L-arabinose]-hederagenin （HXRA）通过激活RAW264.7细胞中NF-
κB和MAPK信号通路，显著增加NO、COX-2、TNF-α、IL-6和ROS的生成，激活巨噬细胞，发挥免疫增强作用（Fig. 9）。为进一步研究
N. sativa作为具有免疫调节作用的营养补充剂提供理论基础。
该系列研究先后发表4 篇论文：
1. Jing Dong, Qiongxin Liang, Yun Niu, Shengjun Jiang, Li Zhou, Jinmei Wang, Changyang Ma, Wenyi Kang; Effects of Nigella sativa seed polysaccharides on type 2 diabetic mice and gut microbiota. International Journal of Biological Macromolecules 2020;159:725-738 https://doi.org/10.1016/j.ijbiomac.2020.05.042 二区 IF 6.953
2. Yun Niu, Li Zhou, Lijun Meng, Sitan Chen, Changyang Ma, Zhenhua Liu, Wenyi Kang; Recent Progress on Chemical Constituents and Pharmacological Effects of the Genus Nigella. Evidence-based complementary and alternative medicine, 2020: 6756835 https://doi.org/10.1155/2020/6756835 四区 IF 2.63
3. Niu Y, Wang B, Zhou L, Ma C, Waterhouse GIN, Liu Z, Ahmed AF, Sun-Waterhouse D and Kang W (2021) Nigella sativa: A Dietary Supplement as an Immune-Modulator on the Basis of Bioactive Components. Front. Nutr. 8:722813. https://doi.org/10.3389/fnut.2021.722813 Top 1区 IF 6.576
4. Qiongxin Liang, Jing Dong, Senye Wang, Wenjing Shao , Adel F. Ahmed, Yan Zhang, Wenyi Kang（2021）Immunomodulatory effects of Nigella sativa seed polysaccharides by gut microbial and proteomic technologies. International Journal of Biological Macromolecules 184: 483–496 https://doi.org/10.1016/j.ijbiomac.2021.06.118 二区 IF 6.953
