Clausena Harmandiana root extract attenuated cognitive impairments via reducing amyloid accumulation and neuroinflammation in Aß1-42-induced rats.

BACKGROUND: Alzheimer's disease (AD) pathogenesis is associated with amyloid-ß (Aß)-induced neuroinflammation. In AD, the activation of microglia caused by Aß accumulation is followed by the synthesis and release of pro-inflammatory cytokines, including interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNFα), and ultimately leads to cognitive impairments. Clausena harmandiana (CH) is a medicinal plant in the Rutaceae family and has been used in folk medicine to relieve illnesses such as stomachache and headache, and as a health tonic. Interestingly, CH root extract (CHRE) has several anti-inflammatory and other pharmacological activities, but there are no studies in AD-like animal models. OBJECTIVES: This study aims to evaluate the effects of CHRE on cognitive impairments, increased Aß1-42 protein levels, and neuroinflammation in Aß1-42-induced rats. METHODS: Forty-eight adult male Sprague-Dawley rats (250-300 g) were randomly divided into 6 groups (n = 8) of the sham control, V + Aß, CB + Aß CHRE125 + Aß, CHRE250 + Aß, and CHRE500 + Aß. Sodium carboxymethylcellulose, Celebrex (10 mg/kg BW) and CHRE (125, 250, and 500 mg/kg BW) were given orally or without any treatment for 35 days. On day 21, aggregated Aß1-42 at a concentration of 1 µg/µl were injected into both lateral ventricles (1 µl/side) of all treated rats, while sterilized normal saline were injected to untreated rats. Ten days later, the novel object recognition test was performed to assess their recognition memory. At the end of the test period, an overdose of thiopental sodium (120 mg/kg BW) and transcardial perfusion with 0.9% normal saline solution were used to euthanize all rats. Then Aß1-42 protein levels and the expression of inflammatory markers (CD11b-positive microglia, IL-1ß, and TNFα) were investigated in the cerebral cortex and hippocampus. RESULTS: Pretreatment with CHRE at all doses could attenuate short- and long-term impairments in recognition memory. Additionally, CHRE also inhibited the increase of Aß1-42 protein levels and the expression of inflammatory markers in both brain regions as well as receiving Celebrex. CONCLUSIONS: This suggests that preventive treatment of CHRE might be a potential therapy against cognitive impairments via reducing Aß1-42 protein levels and neuroinflammation caused by Aß1-42.

Characterization and prebiotic properties of pectin polysaccharide from Clausena lansium (Lour.) Skeels fruit.

Pectins have proven to be advantageous for human health as they regulate beneficial microbial communities and enhance immunity. The fruit of Clausena lansium (Lour.) Skeels (Wampee), also referred to as "treasure in fruit", is rich in pectin polysaccharides. In this study, a homogalacturonan-type pectin (CCP2) with a molecular weight of 8.9 × 104 Da and degree of esterification of 42.86% was isolated from Wampee fruit. The gut microbiota regulation and phagocytosis-enhancing properties of CCP2 were examined in vivo and in vitro, respectively. Oral administration of CCP2 dramatically decreased the abundance of Bacteroidetes and increased the abundance of Firmicutes in intestinal bacteria in mice. The content of short-chain fatty acids in the feces also significantly improved. Moreover, CCP2 exhibited excellent phagocytosis-enhancing activities on RAW 264.7 macrophages. These results suggested that CCP2 could be a potential gut microbiota regulator and phagocytosis-enhancer, which could be used in food products to promote health through beneficial manipulation of gut microbiota.

Ethyl acetate extract of Clausena excavata induces growth inhibition of non-small-lung cancer, NCI-H460, cell line via apoptosis.

CONTEXT: Clausena excavata Burm. f is a plant used in folklore medicine for the treatment of various ailments in South East Asia. The plant parts contain chemical components that are cytotoxic to many cancer cells. OBJECTIVE: The study investigated the cytotoxic effects of ethyl acetate, methanol and chloroform C. excavata leaf extracts on the non-small-lung cancer, NCI-H460, cell line. METHODS: Based on the 3-(4,5-dimethylthiazol-2-yl)-2,5,-diphenyltetrazolium bromide (MTT) assay, among extracts, ethyl acetate C. excavata leaf extract (EACE) was the most potent anti-NCI-H460 cells, with IC50 value of 47.1 ± 6.1 µg/ml. The effects of EACE on NCI-H460 cells were also determined by clonogenic, 4', 6-diamidino-2-phenylindole (DAPI), and annexin-V-fluorescein isothiocyanate/propidium iodide-PI flow cytometric assays. Reactive oxygen species (ROS) production and apoptotic gene expressions was determined via flow cytometry and real-time quantitative PCR, respectively. RESULTS: EACE-treated NCI-H460 cells after 48 h underwent apoptosis as evident by loss of cell viability, cell shrinkage, and chromatin condensation. The results also showed EACE mediated increase in ROS production by the NCI-H460 cells. After 48 h treatment, EACE increased the pro-apoptotic BAX and decreased the anti-apoptotic Bcl-2, Survivin and c-Myc gene expressions. CONCLUSIONS: EACE is a potential anti-lung cancer by increasing cancer cell ROS production and apoptosis.

Anticancer and genotoxicity effect of (Clausena lansium (Lour.) Skeels) Peel ZnONPs on neuroblastoma (SH-SY5Y) cells through the modulation of autophagy mechanism.

Nanotechnology is an emerged field to develop the plant mediated metal based nanodrugs by green method. In this current study, the zinc oxide metal based nanoparticles were developed using (Clausena lansium (Lour.) Skeels) Peel aqueous extracts and zinc nitrate. The C.L extract zinc nanoparticleswere indicated by the sharp peak seen at 350 nm utilizing the Ultraviolet-Visible spectroscopy (UV-Vis). The high peaks indicate the presence of phytochemicals and its functional groups in ZnONPs were studied by the Fourier Transform Infrared Spectroscopy (FT-IR). The X-Ray Diffraction analysis (XRD) explores the pattern and structure of ZnONPs as spherical and base-centered monoclinic crystalline shapes. The C.L extract with Zn nanoparticles were spherical in nature and the size of the synthesized particles were about 28.42 nm respectively. The autophagy (Beclin-1, LC3-I, LC3-II and ATG4B) and apoptotic (Bax, Bcl-2 and Caspase-3) proteins were regulated by the treatment with ZnONPs in SH-SY5Y neuroblastoma cells. The DNA loss or damage was occurred in the ZnONPs treatment and it was performed using Comet assay. The ZnONPs treatment generates the ROS in the cells and decreased its stability and viability. Addition of NAC prevents ROS in the cultured SH-SY5Y cells and prevents the cells from the apoptosis. We concluded that the ZnONPs potentially kills the neuroblastoma cells by producing the intracellular ROS.

Increased Carbazole Alkaloid Accumulation in Clausena harmandiana Callus Culture by Treatments of Biocontrol Agent, Trichoderma harzianum and Bacillus subtilis.

Clausena harmandiana (Pierre) Guillaumin is the Thai medicinal plant that possessed several pharmacological activities. The main active constituents of this plant are the carbazole alkaloids, isolated from wild plants. However, the in vitro culture for production of carbazole alkaloids from this plant has never been reported. Therefore, we aimed to develop callus culture of C. harmandiana elicited with two biotic elicitors, Trichoderma harzianum and Bacillus subtilis, as a sustainable source of carbazole alkaloids. The callus treated with living B. subtilis (BL) at 0.1 and 1% (v/v) for 3 days accumulated 5-fold increased level of clausine K. The highest level reached 309.37 ± 34.84 µg/g DW. This treatment also showed a significant increase in both total phenolic content and antioxidant capacity, which support the optimum usage of this elicitor. Moreover, only callus treated with 1% (v/v) Trichoderma culture filtrate (CF) showed a significant increase in the total phenolic contents and antioxidant capacity. The correlation analysis also revealed the significant correlation between antioxidant capacity and total phenolic level, total flavonoids, and clausine K but not 7-methoxymukonal. The results from our study suggested the use of C. harmandiana callus with the Bacillus elicitors for high-level production of clausine K.

Highly efficient and concise synthesis of both antipodes of SB204900, clausenamide, neoclausenamide, homoclausenamide and zeta-clausenamide. Implication of biosynthetic pathways of Clausena alkaloids.

The synthesis of both antipodes of N-methyl-N-[(Z)-styryl]-3-phenyloxirane-2-carboxamide (SB204900), clausenamide, neoclausenamide, homoclausenamide and zeta-clausenamide have been accomplished using (2S,3R)- and (2R,3S)-3-phenyloxirane-2-carboxamides as the starting materials, and SB204900 was found to be a common precursor to other N-heterocyclic clausena alkaloids. Mediated by Brønsted acids under different conditions, for example, SB204900 underwent efficient and diverse alkene-epoxide cyclization, enamide-epoxide cyclization and arene-epoxide cyclization reactions to produce the five-membered N-heterocyclic neoclausenamide, its 6-epimer, the six-membered N-heterocyclic homoclausenamide and the eight-membered N-heterocyclic zeta-clausenamide, respectively, in good to excellent yields. Regiospecific oxidation of neoclausenamide and its 6-epimer afforded neoclausenamidone. Enolization of neoclausenamidone in the presence of LiOH and the subsequent protonation under kinetic conditions at -78 degrees C led to the epimerization of neoclausenamidone into clausenamidone. Reduction of clausenamidone using NaBH(4) furnished clausenamide in high yield.

[Potential allelopathic effects of Piper nigrum, Mangifera indica and Clausena lansium].

With Piper nigrum, Mangifera indica and Clausena lansium as the donators, this paper studied their potential allelopathic effects on the germination and growth of Zea mays, Glycine max, Cucurbita moschata, Arachis hypogaea, Raphanus sativus, Echinochloa crusgalli, Digitaria sanguinalis and Stylosanthes guianensis. The results showed that the aqueous extracts of these donators could inhibit the germination and growth of Z. mays, G. max, C. moschata, E. crus-galli and D. sanguinalis at high concentration, but stimulate them at low concentration. In rhizosphere soil of P. nigrum and M. indica, the germination and growth of Z. mays L was stimulated, while A. hypogaea was inhibited. The aqueous extracts of the donators were extracted by ethyl acetate and n-butanol, respectively, and the inhibitory activity of both aqueous and n-butanol fractions from P. nigrum and M. indica on Z. mays, R. sativus and S. guianensis was stronger than that of ethyl acetate fraction, indicating that P. nigrum and M. indica contained the allelochemicals with high polarity.

[Changes in water state and soluble protein contents of wampee axes during inducing desiccation tolerance by sucrose preculture].

Progressively increasing sucrose concentration of culture medium could increase desiccation tolerance of wampee [Clausena lansium (Lour.) Skeels] axes. The changes in water state and soluble proteins of axes during acquirement of desiccation tolerance were measured by differential scanning calorimeter (DSC) and SDS-PAGE. The results showed that the cooling and heating thermograms of sucrose-precultured axes were similar to those of the control; but there was a stepwise change in the heating thermograms of sucrose-precultured axes, implying that vitrification might occur in axes. Unfreezable water amounts of wampee axes were measured, the results showed that amounts of unfreezable water of sucrose-precultured axes and control were 25.4% and 25.9% (by the linear regression equation method), or 24.3% and 23.7% (by the heat of ice fusion method) respectively, which were not significantly different. The soluble protein content of sucrose-precultured axes was 68% higher than the control, and SDS-PAGE showed that a 20-kD protein markedly increased in content.