Memory Improvement by Yokukansankachimpihange and Atractylenolide III in the Olfactory Bulbectomized Mice

Alzheimer’s Disease (AD) shows cognitive dysfunction as core symptoms and Behavioral and Psychological Symptoms of Dementia (BPSD). Since acetylcholine nerve system derived from septum is collapsed in the AD patients, we have used Olfactory Bulbectomized (OBX) mice whose cholinergic system is largely impaired in the septum. Recently, Yokukansankachimpihange (YKH), a traditional Japanese Kampo medicine has used for BPSD in addition to improve cognitive dysfunction in AD patients. However the essential components for cognition and BPSD improvement and their mechanism are largely unknown. In present study, we found that Atractylenolide III (Aen-III), one of the components of YKH, improved cognitive deficits and depression in the OBX mice. OBX mice were orally administered with Aen-III (1.0 and 3.0 mg/kg) and YKH extracts daily for 18 days. Like YKH extracts, the Aen-III treatments ameliorated cognitive deficits and depression-like behavior observed in OBX mice. Importantly, Aen-III administration significantly restored the decreases in Ca2+/calmodulin-dependent protein kinase II (CaMKII) autophosphorylation and phosphorylation of Ca2+/calmodulin-dependent protein kinase IV (CaMKIV) and cyclic AMP response element binding protein (CREB). The restoration of CaMKII and CaMKIV signaling is closely related to the increased BDNF levels. Furthermore, ATP reduction in OBX mice was rescued by Aen-III (3.0 mg/kg) and YKH (1000 mg/kg) treatment. In summary, Aen-III as a component of YKH ameliorates cognitive dysfunctions and depression via restoring CaMKII and CaMKIV signaling.

Atractylenolide Ⅰ protects against lipopolysaccharide-induced disseminated intravascular coagulation by anti-inflammatory and anticoagulation effect

Objective:To investigate whether atractylenolide Ⅰ(ATL-Ⅰ) has protective effect on lipopolysaccharide(LPS)-induced disseminated intravascular coagulation(DIC) in vivo and in vitro,and explore whether NF-κB signaling pathway is involved in ATL-Ⅰ treatment.Methods:New Zealand white rabbits were injected with LPS through marginal ear vein over a period of 6h at a rate of 600 μg/kg(10 mL/h).Similarly,in the treatment groups,1.0,2.0,or 5.0 mg/kg ATL-Ⅰ were given.Both survival rate and organ function were tested,including the level of alanine aminotransferase(ALT),blood urine nitrogen(BUN),and TNF-α were examined by ELISA.Also haemostatic and fibrinolytic parameters in serum were measured.RAW 264.7 macrophage cells were administered with control,LPS,LPS + ATL-Ⅰ and ATL-Ⅰ alone,and TNF-α,phosphorylation(P)-IκBα,phosphorylation(P)-NF-κB(P65) and NF-κB(P65) were determined by Western blot.Results:The administration of LPS resulted in 73.3%mortality rate,and the increase of serum TNF-α,BUN and ALT levels.When ATL-Ⅰ treatment significantly increased the survival rate of LPS-induced DIC model,also improved the function of blood coagulation.And protein analysis indicated that ATL-Ⅰ remarkably protected liver and renal as decreasing TNF-α expression.In vitro,ATL-Ⅰ obviously decreased LPS-induced TNF-αproduction and the expression of P-NF-κB(P65),with the decrease of P-IκBα.Conclusions:ATL-Ⅰ has protective effect on LPS-induced DIC,which can elevate the survival rate,reduce organ damage,improve the function of blood coagulation and suppress TNF-α expression by inhibiting the activation of NF-κB signaling pathway.

Identification of an anticancer compound against HT-29 cells from Phellinus linteus grown on germinated brown rice

Objective:To isolate and identify the anticancer compound against proliferation of human colon cancer cells from ethyl acetate(EtOAc)extract ol Phellinus linteus grown on germinated brown rice(PB).Methods:EtOAc extract of PB was partitioned with n-hexane,EtOAc,and water-saturated n-butanol.Anticancer compound of n-hexane layer was isolated and identified by HPLC and NMR,respectively.Cytotoxicity against HT-29 cells was tested by SRB assay.Results:The n-hexane layer obtained after solvent fractionation of PB EtOAc extracts showed a potent anticancer activity against the HT-29 cell line.Atractylenolide I,a eudesmane-type sesquiterpene lactone,a major anticancer substance of PB,was isolated from the n-hexane layer by silica gel column chromatography and preparative-HPLC.This structure was elucidated by one-and two-dimensional NMR spectroscopic data.Atractylenolide I has not been reported in mushrooms or rice as of yet.The isolated compound dose-dependently inhibited the growth of HT-29 human colon cancer cells.Conclusions:Atractylenolide I might contribute to the anticancer effect of PB.

Involvement of mitochondrial apoptotic pathway and MAPKs/NF-κB inflammatory pathway in the neuroprotective effect of atractylenolide Ⅲ in corticosterone-induced PC12 cells

Atractylenolide Ⅲ(ATL-Ⅲ), a sesquiterpene compound isolated from Rhizoma Atractylodis Macrocephalae, has revealed a number of pharmacological properties including anti-inflammatory, anti-cancer activity, and neuroprotective effect. This study aimed to evaluate the cytoprotective efficiency and potential mechanisms of ATL-Ⅲ on corticosterone injured rat phaeochromocytoma(PC12) cells. Our results demonstrate that ATL-Ⅲ increases cell viability and reduces the release of lactate dehydrogenase(LDH). The results suggest that ATL-Ⅲ protects PC12 cells from corticosterone-induced injury by inhibiting the intracellular Ca^(2+) overloading, inhibiting the mitochondrial apoptotic pathway and modulating the MAPK/NF-κB inflammatory pathways. These findings provide a novel insight into the molecular mechanism by which ATL-Ⅲ protected the PC12 cells against corticosterone-induced injury for the first time. Our results provide the evidence that ATL-Ⅲ may serve as a therapeutic agent in the treatment of depression.

Inhibition of ASCT2 induces hepatic stellate cell senescence with modified proinflammatory secretome through an IL-1α/NF-κB feedback pathway to inhibit liver fibrosis

Senescence of activated hepatic stellate cells(aHSCs) is a stable growth arrest that is implicated in liver fibrosis regression.Senescent cells often accompanied by a multi-faceted senescence-associated secretory phenotype(SASP).But little is known about how alanine-serine-cysteine transporter type-2(ASCT2),a high affinity glutamine transporter,affects HSC senescence and SASP during liver fibrosis.Here,we identified ASCT2 is mainly elevated in aHSCs and positively correlated with liver fibrosis in human and mouse fibrotic livers.We first discovered ASCT2 inhibition induced HSCs to senescence in vitro and in vivo.The proinflammatory SASP were restricted by ASCT2 inhibition at senescence initiation to prevent paracrine migration.Mechanically,ASCT2 was a direct target of glutaminolysisdependent proinflammatory SASP,interfering IL-1α/NF-κB feedback loop via interacting with precursor IL-1α at Lys82.From a translational perspective,atractylenolide Ⅲ is identified as ASCT2 inhibitor through directly bound to Asn230 of ASCT2.The presence of -OH group in atractylenolide Ⅲ is suggested to be favorable for the inhibition of ASCT2.Importantly,atractylenolide Ⅲ could be utilized to treat liver fibrosis mice.Taken together,ASCT2 controlled HSC senescence while modifying the proinflammatory SASP.Targeting ASCT2 by atractylenolide Ⅲ could be a therapeutic candidate for liver fibrosis.

Establishment of the model of white blood cell membrane chromatography and screening of antagonizing TLR_4 receptor component from Atractylodes macrocephala Koidz

A model of white blood cell membrane chromatography (WB-CMC) was established to screen active component from Atractylodes macrocephala Koidz. The component can antagonize Toll-like receptor 4 (TLR4) and inhibit inflammatory reaction. In the model of WB-CMC, cell membrane stationary phase (CMSP) was prepared by immobilizing the rabbit white blood cell membrane (WBCM) onto the surface of silica carrier and taxinol was used as a model molecule. The active component which can act on WBCM and its receptor (such as TLR4) as an effective target in A. macrocephala was determined by using a replacement experiment. The anti-inflammatory effects of the active component were tested by using pharmacological methods in vivo. The results indicated that the re-tention characteristics of atractylenolide I as active component was similar to that of taxinol in the model of WB-CMC. And so, atractylenolide I acted on the WBCM and TLR4 and its anti-inflammatory activity was related with antagonizing TLR4. Therefore, the interaction between the active component and WBCM and its receptor can be simulated by the model of WB-CMC in vitro. This model can be used to screen active components and to study effective characteristics for acting on definite targets.

Evaluating the Impact of Trichoderma brevicompactum 31636 on Root Rot of Atractylodes macrocephala and the Fungal Community in the Rhizosphere Soil

Background:Trichoderma brevicompactum can control of plant diseases,but there are few studies about the control of root rot by T.brevicompactum 31636 and its biocontrol mechanism.Objective:To explore the impact of T.brevicompactum 31636 on root rot and the structure of fungal communities of rhizosphere soil.Methods:The effect of T.brevicompactum 31636 on controlling root rot was assessed,the growth of plants was observed by pot experiments,and the fungal structure was detected by high-throughput sequencing analysis.Results:Compared with the CK(healthy plants)group,the plant height,fresh weight,and dry weight increased by 55.3%,48.55%and 57.8%,respectively.While the concentration of atractylenolideⅠ,Ⅱ,Ⅲ,and atractylone increased by 19.35%,16.96%,89.42%and 89.32%,respectively.The results of pot experiments indicated that T.brevicompactum 31636 could significantly improve the survival rate of Atractylodes macrocephala.The results of principal coordinates analysis(PCoA)showed that the sample of CK and Fo+Th(adding Fusdrium oxysporum before T.brevicompactum 31636)group on one side,and samples of Th and Fo group on the other.Furthermore,among the top 20 dominant fungi detected,the fungal genus with 1%increase of relative abundance included Trichoderma(1%),Sordariomycetes(1.5%),Mortierella(1.3%),Penicillium(1%),meanwhile the fungal genus with 1%decrease of relative abundance included Fusarium(30%)and Pleosporales(1%).Conclusion:In summary,T.brevicompactum 31636 can control the root rot of A.macrocephala by regulating of the soil fungal community and inhibiting pathogen.