The Anti-inflammatory Effects of Lignan Glycosides from Cistanche tubu- losa stems on LPS/IFN-gamma-induced RAW264.7 Macrophage Cells via PI3K/ AKT Pathway

Guo, Jinhan1,2; Tang, Shuming3; Miao, Yuyang2,4,5; Ge, Lanlan2,4,5; Xu, Junfa6; Zeng, Xiaobin2,4,5,7

2021

10.2174/1389201021999201124151426

CURRENT PHARMACEUTICAL BIOTECHNOLOGY   影响因子和分区

1389-2010

1873-4316

Background: Cistanche tubulosa is a tonic in traditional Chinese medicines and has a broad spectrum of biological activity, including anti-inflammatory. However, the anti-inflammatory major constituents of C. tubulosa and their underlying mechanisms are still unknown. Objective: The aim of the current study was to explore the separation and structural characterization of lignan glycosides from C. tubulosa (Schenk) Wight., their anti-inflammatory activity and the underlying mechanism. Materials and Methods: Fractionation and isolation of the 85% EtOH extract of C. tubulosa (Schenk) Wight. were carried out and the primary ingredients lignan glycosides (1-6) were structurally characterized. CCK8 methods were used to evaluate the cytotoxic effect of lignan glycosides (1-6). Effects of lignan glycosides (1-6) on NO production in LPS/IFN-gamma-induced RAW264.7 macrophages cells were measured using Griess reagent by reaction with nitrite. The mRNA expression levels of iNOS, COX-2, IL-1 beta, IL-6, TNF-a, and TGF-beta treated RAW264.7 cells with various concentrations (0, 25 and 50 mu g/ml) of lignan glycosides (1, 4) in the presence of LPS (10 ng/ml) and IFN-gamma (20 ng/ml) for 24 h were analyzed by quantitative RT-PCR. Also, the protein expressions of iNOS, COX-2, PI3K, AKT, p-AKT and beta-actin were determined using Western blot analysis. A molecular docking study was performed to investigate the interactions between the lignan glycosides and the PI3K using Autodock vina 1.1.2 package. Results: Six lignan glycosides (1-6) were isolated from stems of C. tubulosa. Among them, (+)pinoresinol-4-O-beta-D-glucopyranosyl-(1 -> 6)-beta-D-glucopyranoside (5) and eleutheroside E (6) were firstly isolated from C. tubulosa. Of these lignans, 1 and 4 exhibited pronounced inhibitions on NO production with the values of 33.63 +/- 4.78 and 39.28 +/- 5.52 % at 50 mu g/ml, respectively. Additionally, LPS/IFN-gamma-induced expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-1 beta (IL-1 beta), IL-6, and tumor necrosis factor-a (TNF-a) was significantly suppressed by pre-treatment of 1 and 4 in a dose-dependent manner. While 1 and 4 increased the mRNA levels of anti-inflammatory cytokines (TGF-beta). Furthermore, 1 and 4 significantly inhibited the protein levels of PI3K and p-AKT in a dose-dependent manner. Conclusion: Taken together, these results suggest that 1 and 4 play an important role in the attenuation of LPS/IFN-gamma-induced inflammatory responses in RAW264.7 cells and that the mechanisms involve down-regulation of the PI3K/AKT pathway.

Cistanche tubulosa lignan glycosides anti-inflammatory activity structure-activity relationship PI3K AKT

SCI

英语

通讯

National Natural Science Foundation of China[81503221,81903760,81903914] ; Shenzhen Fundamental Research and Discipline Layout Project["JCYJ20170413093108233","JCYJ20180228164437735","JCYJ20190806151816859"] ; China Postdoctoral Science Foundation["2018M633290","2019M653300","JCYJ20170307100726777"]

Biochemistry & Molecular Biology ; Pharmacology & Pharmacy

Biochemistry & Molecular Biology ; Pharmacology & Pharmacy

WOS:000663628700011

BENTHAM SCIENCE PUBL LTD

PHARMACOLOGY & TOXICOLOGY

Web of Science

1.Jinan Univ, Shenzhen Peoples Hosp, Clin Med Coll 2, Ultrasonog Dept,Longhua Branch, Shenzhen 518020, Guangdong, Peoples R China
2.Southern Univ Sci & Technol, Affiliated Hosp 1, Shenzhen 518020, Guangdong, Peoples R China
3.Jinan Univ, Shenzhen Peoples Hosp, Clin Med Coll 2, Dept Clin Lab, Shenzhen 518020, Guangdong, Peoples R China
4.Jinan Univ, Shenzhen Peoples Hosp, Clin Med Coll 2, Ctr Lab,Longhua Branch, Shenzhen 518020, Guangdong, Peoples R China
5.Jinan Univ, Shenzhen Peoples Hosp, Clin Med Coll 2, Dept Infect Dis, Shenzhen 518020, Guangdong, Peoples R China
6.Guangdong Med Coll, Inst Lab Med, Guangdong Prov Key Lab Med Mol Diagnost, Dongguan 523808, Guangdong, Peoples R China
7.Shenzhen Univ, Med Sch, Guangdong Prov Key Lab Reg Immun & Dis, Shenzhen 518037, Guangdong, Peoples R China

Guo, Jinhan,Tang, Shuming,Miao, Yuyang,et al. The Anti-inflammatory Effects of Lignan Glycosides from Cistanche tubu- losa stems on LPS/IFN-gamma-induced RAW264.7 Macrophage Cells via PI3K/ AKT Pathway[J]. CURRENT PHARMACEUTICAL BIOTECHNOLOGY,2021,22(10).

Guo, Jinhan,Tang, Shuming,Miao, Yuyang,Ge, Lanlan,Xu, Junfa,&Zeng, Xiaobin.(2021).The Anti-inflammatory Effects of Lignan Glycosides from Cistanche tubu- losa stems on LPS/IFN-gamma-induced RAW264.7 Macrophage Cells via PI3K/ AKT Pathway.CURRENT PHARMACEUTICAL BIOTECHNOLOGY,22(10).

Guo, Jinhan,et al."The Anti-inflammatory Effects of Lignan Glycosides from Cistanche tubu- losa stems on LPS/IFN-gamma-induced RAW264.7 Macrophage Cells via PI3K/ AKT Pathway".CURRENT PHARMACEUTICAL BIOTECHNOLOGY 22.10(2021).