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The present work studies the immunomodulatory potential of nervonic acid (NA) on the growth and proliferation of murine RAW264.7 macrophages. The immunomodulatory effects of NA were assessed through the evaluation of nitric oxide (NO) levels, cytokine secretion, phagocytic activity, reactive oxygen species (ROS) generation, and nuclear factor-κB (NF-κB) signaling pathway of RAW264.7 macrophages. The methodology used included a cell counting kit-8 (CCK-8) technique, enzyme-linked immunosorbent assay (ELISA), neutral red phagocytosis test, flow cytometry, western blotting, and immunofluorescence, respectively. Obtained outcomes pointed to NA as a potential promoter of the production of NO, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and ROS in RAW264.7 cells. Meanwhile, the expression of iNOS, TNF-α, IL-1β, and IL-6 mRNA was upregulated. Besides, the expression of TLR4 protein and the phosphorylation level of NF-κB were increased in RAW264.7 cell lines in a concentration-dependent manner when compared against the control. As a conclusion, NA can activate the NF-κB signaling pathway through the TLR4 signaling pathway, which has immunomodulatory potential. [ABSTRACT FROM AUTHOR]

The root of Croton crassifolius has been used as a traditional Chinese medicine (TCM), called Radix Croton Crassifolius, and commonly known as "Ji Gu Xiang" in Chinese. Its medicinal value has been recorded in several medical books or handbooks, such as "Sheng Cao Yao Xing Bei Yao", "Ben Cao Qiu Yuan" and "Zhong Hua Ben Cao". It has been traditional employed for treating sore throat, stomach-ache, rheumatism and cancer. At present, there are limited studies on the evaluation of low-polarity extracts of roots in C. crassifolius. Consequently, the aim of this study was to evaluate the antitumor effect of the low-polarity extract of C. crassifolius root. Extracts were obtained by supercritical fluid extraction. The extracts were tested for antitumor effects in vitro on several cancer cell lines. A CCK-8 kit was used for further analysis of cell viability. A flow cytometer and propidium iodide staining were used to evaluate the cell cycle and apoptosis. Hoechst staining, JC-1 staining and the fluorescence probe DCFH-DA were used to evaluate apoptotic cells. Molecular mechanisms of action were analyzed by quantitative RT‒PCR and Western blotting. Immunohistochemistry was used for the evaluation of xenograft tumors in male BALB/c mice. Finally, molecular docking was employed to predict the bond between the desired bioactive compound and molecular targets. Eleven diterpenoids were isolated from low-polarity C. crassifolius root extracts. Among the compounds, chettaphanin II showed the strongest activity (IC 50 = 8.58 μM) against A549 cells. Evaluation of cell viability and the cell cycle showed that Chettaphanin II reduced A549 cell proliferation and induced G2/M-phase arrest. Chttaphanin II significantly induced apoptosis in A549 cells, which was related to the level of apoptosis-related proteins. The growth of tumor tissue was significantly inhibited by chettaphanin II in experiments performed on naked mice. The antitumor mechanism of chettaphanin II is that it can obstruct the mTOR/PI3K/Akt signaling pathway in A549 cells. Molecular docking established that chettaphanin II could bind to the active sites of Bcl-2 and Bax. Taken together, the natural diterpenoid chettaphanin II was identified as the major antitumor active component, and its potential for developing anticancer therapies was demonstrated for the first time by antiproliferation evaluation in vitro and in vivo. [Display omitted] • Eleven diterpenoids were obtained from the SFE extract of C. crassifolius roots. • Chettaphanin II showed the strongest antitumor activity against A549 cells. • Chettaphanin II inhibited A549 cell proliferation and arrested cells in G2/M-phase. • Chettaphanin II induced A549 cell apoptosis by inhibiting PI3K/AKT/mTOR pathway. • Chettaphanin II significantly inhibited the growth of tumor tissue in nude mice. [ABSTRACT FROM AUTHOR]

Astragali Radix (AR) is the dry root of the leguminous plants Astragalus membranaceus (Fisch) Beg. var. mongholicus (Beg) Hsiao, and Astragalus membranaceus (Fisch) Bge., being used as a medicinal and edible resource. AR is used in traditional Chinese medicine prescriptions to treat hyperuricemia, but this particular effect is rarely reported, and the associated mechanism of action is still need to be elucidated. To research the uric acid (UA)-lowering activity and mechanism of AR and the representative compounds through the constructed hyperuricemia mouse and cellular models. In our study, the chemical profile of AR was analysed by UHPLC-QE-MS, as well as the mechanism of action of AR and the representative compounds on hyperuricemia was studied through the constructed hyperuricemia mouse and cellular models. The main compounds in AR were terpenoids, flavonoids and alkaloids. Mice group treated with the highest AR dosage showed significantly lower (p < 0.0001) serum uric acid (208 ± 9 μmol/L) than the control group (317 ± 11 μmol/L). Furthermore, UA increased in a dose-dependence manner in urine and faeces. Serum creatinine and blood urea nitrogen standards, as well as xanthine oxidase in mice liver, decreased (p < 0.05) in all cases, indicating that AR could relieve acute hyperuricemia. UA reabsorption protein (URAT1 and GLUT9) was down-regulated in AR administration groups, while the secretory protein (ABCG2) was up-regulated, indicating that AR could promote the excretion of UA by regulating UA transporters via PI3K/Akt signalling pathway. This study validated the activity, and revealed the mechanism of AR in reducing UA, which provided experimental and clinical basis for the treatment of hyperuricemia with it. [Display omitted] • Astragali Radix (AR) main compounds were terpenoids, flavonoids and alkaloids. • Mice treated with the highest AR dose showed significantly lower serum uric acid (UA). • Reduced creatinine and urea nitrogen indicated that AR could relieve acute hyperuricemia. • The UA reabsorption protein (URAT1 & GLUT9) was down-regulated in AR groups. • AR could promote the excretion of UA by regulating PI3K/Akt signalling pathway. [ABSTRACT FROM AUTHOR]

Scheme 1. Synthesis of nuciferine derivative 1a-1r,2a-2l. Reagents and conditions: (a) 48% HBr solution, glacial acetic acid, 70 °C; (b) Et 3 N, N 2 , CH 2 Cl 2 , 25℃, 1h; (c) NaH, N 2 , DMF, 0 °C, 2h. [Display omitted] • The inhibition effect of compound 1j (104.46 ± 4.07%) was roughly the same as benzbromarone (101.44 ± 6.21%) in UA-induced HK-2 cells model; • Compound 1j inhibited the expression of URAT1 and GLUT9 through repressing PI3K/Akt pathway in HK-2 cells; • Compound 1j can promote the excretion of uric acid and thereby increase the fraction of uric acid excretion in vivo ; • Compound 1j can ameliorate renal inflammation and reduce the level of renal inflammatory factors; • Compound 1j shows nephroprotective effect through regulate TLR4/IκBα/NF-κB signaling pathway; Hyperuricemia is a common metabolic disease with a series of complications. Nuciferine, a typical aporphine alkaloid natural compound extracted from the leaves of Nelumbo nucifera Gaertn., was confirmed to have an antihyperuricemia effect. In the present study, 30 novel nuciferine derivatives were designed and synthesized. The effects of all derivatives on the regulation of URAT1 were studied in a uric acid-induced HK-2 cell model with benzbromarone as a positive control. The results indicated that Compound 1j showed the optimal URAT1 inhibitory activity through repressing PI3K/Akt pathway in HK-2 cells and the inhibitory effect was similar to that of benzbromarone. In addition, in vivo experiments demonstrated that Compound 1j could reduce uric acid levels and ameliorate kidney damage in hyperuricemic mice. On the one hand, Compound 1j could inhibit the expression of URAT1 and GLUT9 to increase the uric acid excretion index. On the other hand, Compound 1j could regulate the TLR4/IκBα/NF-κB signaling pathway to reduce the levels of inflammatory cytokines, thereby alleviating kidney damage. Meanwhile, a molecular docking assay revealed the potential molecular binding power (-9.79 kcal/mol) between Compound 1j and URAT1, which was more tightly bound than the lead compound nuciferine (-7.44 kcal/mol). Based on these results, Compound 1j may be a future drug for the development of new potential antihyperuricemia and nephroprotective drug candidates. [ABSTRACT FROM AUTHOR]