Neuroprotective effects of Fructus Chebulae extracts on experimental models of cerebral ischemia

Gaire, B.P.,Kim, H. Pub. Office, Journal of Traditional Chinese Medici 2014 Journal of traditional Chinese medicine Vol.34 No.1

Objective: To investigate the neuroprotective effects of Fructus Chebulae extract using both in vivo and in vitro models of cerebral ischemia. Methods: As an in vitro model, oxygen glucose deprivation followed by reoxygenation (OGD-R) and hydrogen peroxide (H<SUB>2</SUB>O<SUB>2</SUB>) induced cellular damage in rat pheochromocytoma (PC12) cells was used to investigate the neuroprotective effects of extract of Fructus Chebulae. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to calculate cell survival. For in vivo, occlusion of left middle cerebral artery on rats was carried out as a focal cerebral ischemic model. Results: Fructus Chebulae extract increases the PC12 cell survival against OGD-R and H<SUB>2</SUB>O<SUB>2</SUB> by 68% and 91.4% respectively. Fructus Chebulae also decreases the cerebral infarct volume by 39% and extent of hemisphere swelling from 17% in control group to 10% in Fructus Chebulae treated group. Conclusion: Fructus Chebulae, as a traditional medicine, can rescue the neuronal cell death against ischemia related damage. The possible mechanism for the neuroprotection might be the inhibition of oxidative damages occurring after acute phase of cerebral ischemia.

Identification of Sphingosine 1-Phosphate Receptor Subtype 1 (S1P1) as a Pathogenic Factor in Transient Focal Cerebral Ischemia

Gaire, Bhakta Prasad,Lee, Chi-Ho,Sapkota, Arjun,Lee, Sang Yeul,Chun, Jerold,Cho, Hee Jun,Nam, Tae-gyu,Choi, Ji Woong Springer-Verlag 2018 Molecular neurobiology Vol.55 No.3

<P>Medically relevant roles of receptor-mediated sphingosine 1-phosphate (S1P) signaling have become a successful or promising target for multiple sclerosis or cerebral ischemia. Animal-based proof-of-concept validation for the latter is particularly through the neuroprotective efficacy of FTY720, a non-selective S1P receptor modulator, presumably via activation of S1P(1). In spite of a clear link between S1P signaling and cerebral ischemia, it remains unknown whether the role of S1P(1) is pathogenic or neuroprotective. Here, we investigated the involvement of S1P(1) along with its role in cerebral ischemia using a transient middle cerebral artery occlusion ('tMCAO') model. Brain damage following tMCAO, as assessed by brain infarction, neurological deficit score, and neural cell death, was reduced by oral administration of AUY954, a selective S1P(1) modulator as a functional antagonist, in a therapeutic paradigm, indicating that S1P(1) is a pathogenic mediator rather than a neuroprotective mediator. This pathogenic role of S1P(1) in cerebral ischemia was reaffirmed because tMCAO-induced brain damage was reduced by genetic knockdown with an intracerebroventricular microinjection of S1P(1) shRNA lentivirus into the brain. Genetic knockdown of S1P(1) or AUY954 exposure reduced microglial activation, as assessed by reduction in the number of activated microglia and reversed morphology from amoeboid to ramified, and microglial proliferation in ischemic brain. Its role in microglial activation was recapitulated in lipopolysaccharide-stimulated primary mouse microglia, in which the mRNA expression level of TNF-alpha and IL-1 beta, well-known markers for microglial activation, was reduced in microglia transfected with S1P(1) siRNA. These data suggest that the pathogenic role of S1P(1) is associated with microglial activation in ischemic brain. Additionally, the pathogenic role of S1P(1) in cerebral ischemia appears to be associated with the blood-brain barrier disruption and brain-derived neurotrophic factor (BDNF) downregulation. Overall, findings from the current study clearly identify S1P(1) signaling as a pathogenic factor in transient focal cerebral ischemia, further implicating S1P(1) antagonists including functional antagonists as plausible therapeutic agents for human stroke.</P>

Status, Distribution, Conservation and Use Value of Medicinal and Aromatic Plants (MAPs) in Sagarmatha National Park, Nepal

Gaire, Damodar Institute of Forest Science 2014 Journal of Forest Science Vol.30 No.3

The study attempts to assess the status, distribution, conservation and use value of Medicinal and Aromatic Plant (MAPs) in the Sagarmatha National Park. Altogether 62 species of MAPs belonging to 47 genera and 33 families have been recorded in the study area. 10 species, belonging to 9 families are categorized as the potential species. Out of the these species, most potential in local but threatened species are Allium hypsistum Stearn, Cordyceps sinensis Sacc, Dactylorhiza hatagirea Soo, Nardostachys grandiflora DC, Aconitum orochryseum, Ephedra gerardiana Wall. Ex. Stapf, Swertia multicaulis D. Don, Picrorhiza scrophulariflora Penne, Rheum australe. D. Don, Malva verticillataL and Swertia pedicallata Benerji. By analysis of data using Simpson's diversity index (SI) and Shannon weaver function (H'), there was high diversity (more heterogeneous) MAPs species composition in the Manjo Gate to Large Dobhan. (0.98349). Less diversity (less heterogeneous) MAPs species composition was in Tyanboche to Pangoche (0.90419). Similarly, the Shannon weaver function shows that in plots laying out in Mongla to Phorche are evenly distributed than others However, higher MAPs species (i.e., 31) was found in the way of Manjo Gate to Larja Dobhan than others.

Variation of Medicinal Plants Species Richness along Vertical Gradient in Makawanpur District, Nepal

Damodar Gaire,Lichun Jiang,Vijay Kumar Yadav,Jit Narayan Shah,Sunita Dhungana,Anju Upadhyaya,Shiv Kumar Manjan,Binod Kumar Heyojoo 강원대학교 산림과학연구소 2021 Journal of Forest Science Vol.37 No.2

The research attempted to analyze the medicinal plant species richness in the vertical gradient from lower to the highest elevation of Makawanpur, focusing on the relationship between species richness and elevation which is very important for conservation and management of species diversity. Inventory was carried out in the study area by taking sampling intensity of 0.5% in the effective area. Altogether, 42 sample plots were laid in the field with the help of GIS software maintaining 50 m altitude difference. High species diversity was found in the herbs species whereas shrubs have comparatively low species diversity. The maximum species richness is found in herbs and poles whereas shrubs and trees have relatively low species richness. Research showed that species richness of medicinal plants increased with altitudinal gradient. While analyzing the species richness from 350 to 2,550 m (msl), the highest species richness was received with the elevation ranges from 1,800 m to 2,300 m. There was a positive relationship between species richness and altitudinal gradient in the study area. In addition, we have recorded the high value medicinal plants after 1,800 m altitude and rarely within 1,000 m. Medicinal plants correlated both positive and negative relationships with the increased altitude. The altitudinal response has positively seen except density (n/ha) of Shrubs. Domestication and cultivation of high value medicinal plants should be promoted in community forest including private lands. Training, workshops and awareness programs should be conducted to make people aware about medicinal plants resource utilization, conservation and commercialization of available medicinal plants.

Status, Distribution, Conservation and Use Value of Medicinal and Aromatic Plants (MAPs) in Sagarmatha National Park, Nepal

Damodar Gaire 강원대학교 산림과학연구소 2014 Journal of Forest Science Vol.30 No.3

The study attempts to assess the status, distribution, conservation and use value of Medicinal and Aromatic Plant (MAPs) in the Sagarmatha National Park. Altogether 62 species of MAPs belonging to 47 genera and 33 families have been recorded in the study area. 10 species, belonging to 9 families are categorized as the potential species. Out of the these species, most potential in local but threatened species are Allium hypsistum Stearn, Cordyceps sinensis Sacc, Dactylorhiza hatagirea Soo, Nardostachys grandiflora DC, Aconitum orochryseum, Ephedra gerardiana Wall. Ex. Stapf, Swertia multicaulis D. Don, Picrorhiza scrophulariflora Penne, Rheum australe. D. Don, Malva verticillataL and Swertia pedicallata Benerji. By analysis of data using Simpson’s diversity index (SI) and Shannon weaver function (H’), there was high diversity (more heterogeneous) MAPs species composition in the Manjo Gate to Large Dobhan. (0.98349). Less diversity (less heterogeneous) MAPs species composition was in Tyanboche to Pangoche (0.90419). Similarly, the Shannon weaver function shows that in plots laying out in Mongla to Phorche are evenly distributed than others However, higher MAPs species (i.e., 31) was found in the way of Manjo Gate to Larja Dobhan than others.

S1P1 Regulates M1/M2 Polarization toward Brain Injury after Transient Focal Cerebral Ischemia

( Bhakta Prasad Gaire ),( Young Joo Bae ),( Ji Woong Choi ) 한국응용약물학회 2019 Biomolecules & Therapeutics(구 응용약물학회지) Vol.27 No.6

M1/M2 polarization of immune cells including microglia has been well characterized. It mediates detrimental or beneficial roles in neuroinflammatory disorders including cerebral ischemia. We have previously found that sphingosine 1-phospate receptor subtype 1 (S1P₁) in post-ischemic brain following transient middle cerebral artery occlusion (tMCAO) can trigger microglial activation, leading to brain damage. Although the link between S1P₁ and microglial activation as a pathogenesis in cerebral ischemia had been clearly demonstrated, whether the pathogenic role of S1P₁ is associated with its regulation of M1/M2 polarization remains unclear. Thus, this study aimed to determine whether S1P₁ was associated with regulation of M1/M2 polarization in post-ischemic brain. Suppressing S1P₁ activity with its functional antagonist, AUY954 (5 mg/kg, p.o.), attenuated mRNA upregulation of M1 polarization markers in post-ischemic brain at 1 day and 3 days after tMCAO challenge. Similarly, suppressing S1P₁ activity with AUY954 administration inhibited M1-polarizatioin-relevant NF-κB activation in post-ischemic brain. Particularly, NF-κB activation was observed in activated microglia of post-ischemic brain and markedly attenuated by AUY954, indicating that M1 polarization through S1P₁ in post-ischemic brain mainly occurred in activated microglia. Suppressing S1P₁ activity with AUY954 also increased mRNA expression levels of M2 polarization markers in post-ischemic brain, further indicating that S1P₁ could also influence M2 polarization in post-ischemic brain. Finally, suppressing S1P₁ activity decreased phosphorylation of M1-relevant ERK1/2, p38, and JNK MAPKs, but increased phosphorylation of M2-relevant Akt, all of which were downstream pathways following S1P₁ activation. Overall, these results revealed S1P₁-regulated M1/M2 polarization toward brain damage as a pathogenesis of cerebral ischemia.

Anti-neuroinflammatory and neuroprotective effects of the Lindera neesiana fruit in vitro

Subedi, L.,Gaire, B.P.,Do, M.H.,Lee, T.H.,Kim, S.Y. G. Fischer 2016 Phytomedicine Vol.23 No.8

<P>Background: Lindera neesiana Kurz (Lauraceae), popularly known as Siltimur in Nepal, is an aromatic and spicy plant with edible fruits. It is a traditional herbal medicine widely used for the treatment of diarrhea, tooth pain, headache, and gastric disorders and is also used as a stimulant. Purpose: The aim of the present study was to examine in vitro cytoprotective, anti-neuroinflammatory and neuroprotective potential of an aqueous extract of L. neesiana (LNE) fruit using different central nervous system (CNS) cell lines. Methods: In order to study the neuroprotective potential of LNE, we used three different types of CNS cell lines: murine microglia (BV2), rat glioma (C6), and mouse neuroblastoma (N2a). Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reagent, and prostaglandin E-2 (PGE(2)), tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-6, and nerve growth factor (NGF) release in the culture media was determined using enzyme linked immunosorbent assay (ELISA) kits. Western blot analysis was performed to determine the protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX2), mitogen activated protein kinase (MAPK) family proteins, Bax, B cell lymphoma (BCL)-2, and cleaved caspase 3. Neurite outgrowth was determined using the IncuCyte imaging system. Results: LNE treatment not only reduced nitric oxide (NO) production in a dose-dependent manner, but also significantly reduced proinflammatory cytokines, iNOS and COX-2 production by lipopolysaccharide (LPS) stimulated BV-2 cells. LNE increased the expression of phosphorylated (p)-extracellular signal-regulated kinase (ERK), whereas p-p38 and p- janus kinase (JNK) expression was significantly decreased in activated microglia. Furthermore, LNE increased cell viability of N2a cells, which was accompanied by decreased caspase-3 expression and the ratio of Bax/Bcl2 protein expression as well as increased NGF and neurite outgrowth, suggesting its neuroprotective potential against LPS-induced effects. Additionally, LNE substantially increased nuclear factor erythroid 2-related factor 2 (Nrf2) secretion in N2a cells and inhibited lipid dehydrogenase (LDH) release in H2O2-stimulated BV2 cells demonstrating the strong anti-inflammatory and antioxidant effects of LNE in CNS cell lines. Conclusion: Here we found that water the soluble extract of LNE has promising anti-neuroinflammation and anti-apoptotic properties and identify LNE as a potential natural candidate for neuroprotection. (C) 2016 Published by Elsevier GmbH.</P>

Identifying lysophosphatidic acid receptor subtype 1 (LPA₁) as a novel factor to modulate microglial activation and their TNF-α production by activating ERK1/2

Kwon, Jin Hyun,Gaire, Bhakta Prasad,Park, Se Jin,Shin, Dong-Yoon,Choi, Ji Woong Elsevier 2018 Biochimica et biophysica acta, Molecular and cell Vol.1863 No.10

<P><B>Abstract</B></P> <P>Microglia regulate immune responses in the brain, and their activation is key to the pathogenesis of diverse neurological diseases. Receptor-mediated lysophosphatidic acid (LPA) signaling has been known to regulate microglial biology, but it is still unclear which receptor subtypes guide the biology, particularly, microglial activation. Here, we investigated the pathogenic aspects of LPA receptor subtype 1 (LPA<SUB>1</SUB>) in microglial activation using a systemic lipopolysaccharide (LPS) administration-induced septic mouse model in vivo and LPS-stimulated rat primary microglia in vitro. LPA<SUB>1</SUB> knockdown in the brain with its specific shRNA lentivirus attenuated the sepsis-induced microglia activation, morphological transformation, and proliferation. LPA<SUB>1</SUB> knockdown also resulted in the downregulation of TNF-α, at both mRNA and protein levels in septic brains, but not IL-1β or IL-6. In rat primary microglia, genetic or pharmacological blockade of LPA<SUB>1</SUB> attenuated gene upregulation and secretion of TNF-α in LPS-stimulated cells. In particular, the latter was associated with the suppressed TNF-α converting enzyme (TACE) activity. We reaffirmed these biological aspects using a BV2 microglial cell line in which LPA<SUB>1</SUB> expression was negligible. LPA<SUB>1</SUB> overexpression in BV2 cells led to significant increments in TNF-α production upon stimulation with LPS, whereas inhibiting LPA<SUB>1</SUB> reversed the production. We further identified ERK1/2, but not p38 MAPK or Akt, as the underlying effector pathway after LPA<SUB>1</SUB> activation in both septic brains and stimulated microglia. The current findings of the novel role of LPA<SUB>1</SUB> in microglial activation along with its mechanistic aspects could be applied to understanding the pathogenesis of diverse neurological diseases that involve microglial activation.</P> <P><B>Highlights</B></P> <P> <UL> <LI> LPA<SUB>1</SUB> signaling triggers microglial activation in LPS-induced septic mouse brain and LPS-stimulated rat primary microglia. </LI> <LI> LPA<SUB>1</SUB> signaling upregulates TNF-α production through the activation of TNF-α converting enzyme. </LI> <LI> ERK1/2 is identified as the underlying effector pathway responsible for the TNF-α production by LPA<SUB>1</SUB> in activated microglia. </LI> </UL> </P>

갈근 에탄올추출물의 국소뇌허혈 모델에 대한 뇌신경보호 효과

송정빈,최진규,이동헌,Bhakta Prasad Gaire,이창원,부영민,최호영,김호철 대한본초학회 2012 大韓本草學會誌 Vol.27 No.6

Objectives: The purpose of this study was to evaluate the neuroprotective effect of Pueraria lobata extract on focal cerebral ischemia in mice. Methods: Focal cerebral ischemia was induced by occlusion of the right middle cerebral artery using the intraluminal filament model. ICR male mice underwent 90 minutes of middle cerebral artery occlusion (MCAo) followed by 24 hours of reperfusion. Mice were administered Pueraria lobata extract orally at the dose of 300mg/kg just prior to reperfusion. Rotarod test and balance beam test were practiced to assess sensory-motor function 23 hours after MCAo. In rotarod test, the latency to fall on the accelerating rotarod was recorded for 5 min. In balance beam test, the score was graded according to number of slips and latency to cross. The infarct volume was measured 24 hours after MCAo using 2% 2,3,5-triphenyltetrazolium chloride (TTC) staining. Results: Pueraria lobata extract treated group showed significant reduction in infarct volume by 27.3% compared to control group (p<0.05). In rotatod test, it also showed significant extension of latency time compared to control group (67.82±15.08 vs. 5.62±1.06, p<0.001). In contrast to performance in rotarod test, that in balance beam test did not improve with Pueraria lobata extract treatment. Conclusions: We conclude that Pueraria lobata extract has a significant neuroprotective effect and reduces damage of sensory-motor function in MCAo model. These findings suggest that Pueraria lobata could be a potent neuroprotective agent.

한약복합추출물 HT008의 제조방법에 따른 항염증 효능 및 성분함량 비교연구

이동헌 ( Dong Hun Lee ),송정빈 ( Jung Bin Song ),송미경 ( Mi Kyung Song ),( Bhakta Prasad Gaire ),박주연 ( Ju Yeon Park ),김애홍 ( Ai Hong Jin ),최호영 ( Ho Young Choi ),김호철 ( Ho Cheol Kim ) 대한본초학회 2013 大韓本草學會誌 Vol.28 No.4

Objectives : The biological activities and compound contents of herbal medicine vary depending on manufacturing processes. In this study, we compared anti-inflammatory effects and compound contents of three kinds of multi-herbal extract HT008 produced by different manufacturing processes in order to determine chemical and biological equivalence. Methods : HT008 was produced by three different manufacturing methods: 1. Freeze dried extract of Eleutherococcus senticosus, Scutellaria baicalensis and Angelica sinensis (HT008 FD), 2. Spray dried extract of E. senticosus and S. baicalensis combined with reflux extract of A. sinensis (HT008 SD), 3. Spray dried extract of E. senticosus and S. baicalensis combined with supercritial fluid extract of A. sinensis (HT008 SF). Anti-inflammatory effects were evaluated using acetic acid induced pain model and λ-carageenan induced paw edema model. Compound contents were evaluated by HPLC quantitative analysis of standard compounds of HT008, eleutheroside E, baicalin, z-ligustilide. Results : HT008 FD, HT008 SD and HT008 SF significantly decreased acetic acid induced pain index and λ -carrageenan induced paw edema volume compared with that of control group. There was no significant difference in efficacy among the HT008 FD, HT008 SD and HT008 SF. Standard compound contents of HT008 FD, HT008 SD and HT008 SF were quantified within the range of Korean pharmacopoeia or other research. Conclusions : Three different manufacturing methods of multi-herbal extracts have been developed without noticeable difference in the efficacy or compound contents. The results might be used to establish manufacturing process and industrialization of herbal extracts.