Neuroprotective Effect Of Extract Research Articles

A Mini-Review on the Neuroprotective Effects of Extracts and Metabolites of Momordica charantia

A Mini-Review on the Neuroprotective Effects of Extracts and Metabolites of Momordica charantia

Lipid peroxidation reduction and hippocampal and cortical neurons protection against ischemic damage in animal model using Stellaria media

This study was aimed to determine the neuroprotective influence of Stellaria media in terms of restoring normal state of the rat’s hippocampus and cortex after oxidative insult caused by in vitro ischemia and reperfusion. Cell viability and membrane integrity were assessed using MTT and lactate dehydrogenase (LDH) assay, respectively. Ischemic insult was introduced in the rat brain’s hippocampal and cortical slivers by exposing oxygen and glucose deficiency (OGD) for 2 h, followed by 1 h of re-perfusion. Cellular oxidative stress levels were quantified by incorporating 2ʹ,7ʹ-dichlorofluorescein diacetate fluorescent probes. Additionally, the lipid peroxidation was assessed using TBARS assay. Findings revealed significant neuroprotection against OGD-induced mitochondrial impairment at 40 µg/mL of S. media in rat’s hippocampal and cortical slices. The LDH levels were decreased significantly (P < 0.001) during pre-incubation and reoxygenation periods using varied concentrations of S. media extract. Cellular oxidative stress levels results showed significant (P < 0.001) reduction in dichlorofluorescein fluorescence in slices homogenate of hippocampus and cortex using S. media extract. The lipid peroxidation assay results showed decreased (P < 0.01) levels of malondialdehyde in liver tissues of treated rats treated (200 mg/kg body weight) when compared to the ischemic animal. In summary, findings clearly indicated the neuroprotective effects of extract against in vitro ischemia in brain hippocampal and cortex slivers. S. media could undoubtedly be utilized as a healing agent in preventing neuronal cells’ loss during is chemic-reperfusion process.

제주도 탈염 용암 해수를 이용하여 수경재배한 새싹인삼 추출물의 In Vitro 신경독성에 대한 신경보호 효과

본 연구에서는 유용한 미네랄 성분을 다량 함유하고 있어 산업적 가치가 높은 제주 용암 해수에서 탈염 과정을 거쳐 만들어진 탈염 용암 해수를 이용하여 수경재배한 새싹인삼추출물 건조시료를 Q-Orbitrap으로 성분을 규명하고, HPLC로 기준 및 시험방법을 설정하여 표준화했으며 ICP-AES를 이용하여 무기물 함량을 분석하였다. 시험결과에서 주요 ginsenoside Rb1, Rg1 및 Na, Mg, Ca, Sr, V와 같은 미네랄 함량이 증가함을 확인했으며 이에 대한 효능을 입증하기 위해서 제주도 탈염 용암 해수를 이용해 수경재배한 새싹인삼 추출물에 대한 신경세포사멸과 신경세포 염증 반응 억제, 신경전달물질 및 시냅스 가소성 회복을 통한 기억력 개선에 대해 in vitro 효능검증을 수행하였다. 제주도 탈염 용암 해수 수경재배 새싹인삼 추출물에 대한 동결 건조를 진행 후 ginsenoside 함량 분석을 진행한 결과에서 ginsenoside Rb1(11.5 mg/g), Rg1(6.85 mg/g), Re(19.2 mg/g)의 합이 37.55 mg/g으로 측정되었고, 이는 식약처 건강기능식품 인삼 기능성분의 함량기준인 Rb1, Rg1의 합이 0.8~34 mg/g인 것과 비교했을 때, 탈염 용암 해수로 수경재배한 제주새싹인삼 동결 건조물은 개별 인정형 원료로서의 충분한 요건을 갖춘 것으로 사료된다. 또한 제주도 탈염 용암 해수 수경재배 새싹인삼 추출물 건조시료에 대한 세포독성 및 효능평가 결과에서 제주도 탈염 용암 해수 수경재배 새싹인삼추출물 건조시료는 RA-differentiated SH-SY5Y neuronlike cell에 48시간 동안 처리 시 1,250 μg/mL에서도 세포독성을 보이지 않았으며, 더욱이 Aβ1-42 oligomer(10 μM)를 72시간 동안 처리하여 세포독성을 유도한 세포실험 모델에서 비처리 대조군과 비교하여 제주도 탈염 용암 해수 수경재배 새싹인삼 추출물 건조시료를 처리한 실험군에서 세포사멸 관련 바이오마커들인 cleaved/total caspase-3 ratio, cleaved/total PARP, Bax/Bcl-2 ratio를 감소시켜 Aβ1-42 oligomer에 의한 신경세포사멸을 억제하는 효과를 보였다. 그리고 RA-differentiated SH-SY5Y neuron-like cell에 H₂O₂(300 μM)를 처리하여 신경세포 염증 반응을 유도한 세포실험 모델에서 비처리 대조군과 비교하여 제주도 탈염용암 해수 수경재배 새싹인삼 추출물 건조시료를 처리한 실험군에서 염증 반응과 관련된 NF-κB p65 단백질의 nuclear translocation 억제, COX-2 단백질 발현 감소, MAPK (ERK1/2, SAPK/JNK, p38 MAPK) 인산화를 감소시켜 H₂O₂에 의한 신경세포 염증 반응을 억제하는 효과를 보였다. 한편 RA-differentiated SH-SY5Y neuron-like cell에 Aβ1-42 oligomer(1 μM)를 처리하여 신경전달물질 및 시냅스 가소성 감소를 유도한 세포실험 모델에서 비처리 대조군과 비교하여 제주도 탈염 용암 해수 수경재배 새싹인삼 추출물 건조시료를 처리한 실험군에서 시냅스 가소성과 관련된 CREB 전사인자의 인산화를 통한 활성화로 신경전달물질인 BDNF 발현이 증가하는 효과를 보였으며, AChE 활성을 저해함으로써 Aβ1-42 oligomer에 의한 신경전달물질 및 시냅스 가소성 감소를 억제하는 효과를 보였다. 그러므로 제주도 탈염 용암 해수를 이용한 새싹인삼은 질환 특이적인 성분의 증가를 높여줌으로써 산업적 이용가치가 높을 것으로 판단되며, 이를 활용한 다른 약용작물 및 특용작물의 생산을 통한 건강기능식품 원료개발에도 충분히 활용 가능할 것으로 사료된다.

Neuroprotective Effects of Extracts from the Radix Curcuma aromatica on H2O2-induced Damage in PC12 Cells.

The model of oxidative stress damage was established by treatment of 400 µM H2O2 on PC12 to induce cell damage. After the treatment of ECWs for 24 h, the cell viability, LDH, SOD, CAT and GSH were measured to evaluate the neuroprotection of ECAs on that model. The potential action mechanism was studied by measurement of level of ROS, cell apoptosis rate, mitochondrial membrane potential (MMP), morphologic change, the intracellular Ca2+ content (F340/F380) and the expressions of Bcl-2, Bax and Caspase-3. Additionally, the constituents from tested extracts were analyzed by HPLC-DAD-Q-TOF-MS method. Compared with a positive control, Vitamin E, 10 µg/ml of 95% EtOH extract (HCECA) and 75% EtOH extract (MCECA) can markedly increase the rate of cell survival and enhance the antioxidant enzyme activities of SOD, CAT, increase the levels of GSH, decrease LDH release and the level of ROS, attenuate the intracellular Ca2+ overloading, reduce the cell apoptotic rate and stabilize MMP, down-regulate Bcl-2 expression, up-regulate Bax and caspase-3 expression, and improve the change of cell morphology. The chemical analysis showed that diarylheptanoids and sesquiterpenoids are the major chemicals in tested extracts and the former were richer in HCECA and MCECA than others. These findings indicated that the effects of HCECA and MCECA on inhibiting the cells damage induced by H2O2 in PC12 are better than other extracts from the radix of C. aromatica, and the active constituents with neuroprotective effects consisting in those two active extracts are diarylheptanoids.

Comparison of neuroprotective effect of Forsythia suspensa leaf extract and forsythiaside, one of its metabolites

As one of authentic Chinese medicinal materials in Shanxi Province, leaves of Forsythia suspensa (Thunb.) Vahl (F. suspensa, Lianqiao in Chinese) have been used as alternative for the relief or treatment of inflammation. The study aimed to compare the neuroprotective effects of extract from F. suspensa leaves and forsythiaside against inflammation and apoptosis induced by lipopolysaccharide (LPS) on hippocampal slices. The cultured hippocampal slices were treated with extract (5 and 50 μg/mL) and forsythiaside (5 and 50 μg/mL), respectively, followed by LPS treatment for 24 h. The present study showed that 60% ethanol extract of F. suspensa leaves exhibited better neuroprotective effect than equivalent forsythiaside.

잔대 추출물과 이들 분획물들의 항산화 및 뇌신경세포 보호 효과

잔대의 70% 에탄올 추출물은 강한 항산화 효과 및 산화적 스트레스 조건에서 뇌신경세포 사멸 억제 효과가 있었다. 6개의 분획물 중에 잔대 추출물의 클로로포름 분획물(ATCH)은 높은 DPPH 라디칼 소거작용 및 세포 내 활성산소종(ROS) 소거작용을 가지고 있었고 H₂O₂에 대항하여 뇌신경세포(SK-N-SH) 보호 효과가 가장 뛰어났다. 활성이 가장 우수한 클로로포름 분획물을 대상으로 주요 물질을 순수분리한 후 ¹H-NMR, SUP13/SUPC-NMR 그리고 EI mass spectra를 이용하여 화학구조를 분석한 결과 phytosterols(β-sitosterol과 daucosterol)이라는 것을 알 수 있었다. β-Sitosterol과 daucosterol 역시 산화적 스트레스에 대항하여 SK-NSH세포를 보호하는 작용이 있었다. AT-CH 그리고 이것으로부터 분리된 화합물은 p38 경로를 저해하였다. 이들 결과는 AT-CH는 p38 경로를 저해하고 세포 내 ROS 소거작용에 의해 산화적 스트레스로부터 뇌신경세포 보호 효과를 가질 것이라 제안하였다.

Comparison of neuroprotective effects of extract and fractions from Agarum clathratum against experimentally induced transient cerebral ischemic damage

Contexts: Agarum clathratum (Laminariaceae), a typical brown algae, has been identified by National Plant Quarantine Service in Korea. The extract of A. clathratum has antioxidant activities.Objective: We investigated the neuroprotective effects of crude-extract, ethyl acetate (EA)-, n-butanol (BU)-, dichloromethane (DCM)- and n-hexane (Hx)-fractions from A. clathratum on ischemic damage in the gerbil hippocampal CA1 region (CA1) after 5 min of transient cerebral ischemia.Materials and methods: Agarum clathratum was collected in Kangwon province (South Korea) and treated with 95% ethanol. The ethanol extract was suspended in distilled water and subjected to a series of partitions with EA, BU, DCM and Hx. Each of extract and fraction was orally administered with 50 mg/kg once a day for one week before ischemia--reperfusion (I-R).Result: In the crude-extract-, EA- and BU-fraction-treated ischemia groups, we found strong neuroprotection in the CA1 – about 80–89% of CA1 pyramidal neurons survived. However, in the DCM- and Hx-fraction-treated ischemia groups, we did not find any significant neuroprotection. In addition, we observed changes in astrocytes and microglia in the ischemic CA1. In the crude-extract, EA- and BU-fraction-treated ischemia groups, the distribution pattern and activity of the glial cells were similar to that found in the sham group.Discussion: Repeated supplements of crude-extract, EA- and BU-fractions of A. clathratum could protect neurons from I-R injury in the hippocampal CA1 induced by transient cerebral ischemia via decrease of glial activation.

Protective effect of extracts of calyx from Diospyros kaki on H 2 O 2 -induced cytotoxicity in PC12 cells

Oxidative stress is considered a major cause of cellular injuries, including various clinical abnormalities. Here, the neuroprotective effect of extracts from the calyx of Diospyros kaki(DCE), peel of D. kaki (DPE), and flesh of D. kaki (DFE) of Diospyros kaki on oxidative stress-induced apoptosis in PC12 cells was investigated by an 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) reduction assay. The results showed that H2O2significantly decreased cell viability (50.9%), and DCE exerted the highest neuroprotective effect on H2O2-induced cytotoxicity. We identified that treatment with DCE of H2O2-stressed PC12 cells caused dose-dependent suppression of H2O2-induced leakage of lactate dehydrogenase (LDH) as released amount (11.1 to 30.4%); we further verified these findings by observing morphological features. H2O2 also induced severe apoptosis of the PC12 cells, as determined by Hoechst 33342 staining and flow cytometric analysis. DCE (100, 500 µg/ml) exerted a significantly high protective effect on PC12 cells against H2O2-induced cell injury, and the percentage of cells in the sub-G1 phase decreased to 3.7 and 4.6%. These results suggest that DCE can protect cells against H2O2-induced apoptosis, and might be a potential therapeutic agent for treating or preventing neurodegenerative diseases that are related to oxidative stress. Key words: Apoptosis, Diospyros kaki, neurodegenerative disease, neuroprotective effect, oxidative stress.

Neuroprotective Effect of Total and Sequential Extract of Scrophularia striata Boiss. in Rat Cerebellar Granule Neurons Following Glutamate- Induced Neurotoxicity: An In-vitro Study.

Neuroprotective effect of the extract from aerial parts of Scrophularia striata Boiss (Scrophulariaceae) was investigated against glutamate-induced neurotoxicity on cultured rat pups Cerebellar Granule Neurons (CGNs). CGNs from 8 days old Sprague-Dawley rat were prepared and cultured. The experiments were performed after 8 days in culture. The plant was collected from the northeastern part (Ruin region) of Iran and air-dried at room temperature. The total extract was prepared with maceration of prepared powder in ethanol 80% for three times. SEQUENTIAL EXTRACTS WERE OBTAINED USING DRIED AND POWDERED AERIAL PARTS WITH INCREASINGLY POLAR SOLVENTS: petroleum ether, chloroform, ethyl acetate and methanol 80% solution. Cultured cells were exposed to 125 μM of glutamate for 12 h following a 24 h of incubation with test fractions at concentration of 10 mcg/mL. Morphological assay was performed using invert light microscope after fixation and staining with haematoxylin. Neuronal viability was measured using MTT assay. Statistical analysis was done using SPSS software. One way analysis of variance (ANOVA) was performed by Tukey post-hoc test. Values were considered statistically significant when p-value ≤ 0.05. Results of this study showed a significant neuroprotective activity of high polarity methanolic fraction of aerial parts of Scrophularia striata against glutamate-induced neurotoxicity in a dosedependent manner. Treatment with 10 mcg/mL of the fractions showed the best result.

치콘 추출물의 항산화 활성

본 연구는 치콘 추출물의 항산화 효능을 확인하고자 물과 에탄올로 추출하였다. 치콘에 포함된 총 폴리페놀 함량을 측정한 결과, 에탄올 추출물(37.3±5.2 ㎎/GAE/g extract)과 열수 추출물(36.3±1.0 ㎎/GAE/g extract)이 유사한 총 폴리페놀 함량을 포함하고 있었으며, 총 플라보노이드 함량은 물 추출물이 47.0±3.8 ㎎ CE/g extract 그리고 에탄올 추출물이 53.9±5.1 ㎎ CE/g extract를 나타내었다. ABTS를 이용한 라디칼 소거활성, FRAP를 이용한 환원력을 통한 항산화 활성을 측정한 결과에서도 치콘 추출물이 항산화 효과를 나타내었다. 한편, 세포 독성을 살펴보기 위하여 신경세포를 이용하여 MTT assay를 수행한 결과, 세포의 생존율은 1.0 ㎎/㎖의 농도 이상에서는 생존에 영향을 미치지 않았고 신경세포 보호효능 실험에서는 2.5 mM의 H<SUB>2</SUB>O<SUB>2</SUB>로 유발시킨 산화적 손상에 대해 농도 의존적인 신경세포 보호 효과가 있었으며, 항산화 효소 활성을 SOD와 CAT로 분석한 결과 SOD는 0.5 mg/mL 농도에서 95% 이상의 활성과 CAT는 손상그룹에 비해 2배 이상의 활성을 각각 확인하였다. 또한 치콘 추출물이 세포사와 관련이 있는 단백질인 Bax와 Bcl-2의 발현을 조절하는 것을 확인하였다. 이와 같은 결과는 치콘 추출물이 산화적 손상의 억제를 통해 세포를 보호하는 효과가 있는 것으로 사료된다.

땅콩나물 추출물의 신경세포 보호 효과

본 연구는 땅콩나물 추출물이 glutamate가 유도하는 세포독성으로부터 신경세포를 보호하는 효과를 확인하였다. 땅콩나물을 부위별로 전체, 머리, 줄기 부분으로 나누어 methanol로 처리하여 얻어진 각각의 추출물 WME, HME 그리고 SME를 이용하여 glutamate에 의하여 유도된 세포독성에 대한 신경세포보호효과를 관찰하였다. 기지의 신경세포 N18-RE-105 세포주를 이용하여 MTT reduction assay, LDH release assay, 형태학적인 변화 및 apoptosis를 관찰한 결과로부터 HME에서 효율적인 신경세포보호효과를 보였다. 다음으로 HME를 이용하여 hexane, diethyl ether, ethyl acetate, water 층으로 분획하여 신경세포 보호 효과를 확인한 결과, diethyl ether 층에서 가장 높은 활성을 확인할 수 있었다. 그리고 HME의 apoptosis 억제 효과를 확인하기 위하여 flow cytometric analysis를 실시한 결과에서 glutamate 만을 처리하였을 경우 sub-G1기 세포가 58.5%의 확인되었으나 HME를 100 mg/ml 동시 처리하였을 때에는 sub-G1 세포가 9.1%로 감소하여 높은 apoptosis 억제 효과를 확인할 수 있었다. 이상의 결과로부터 땅콩나물 머리 부분 methanol 추출물에는 glutamate에 의한 세포독성으로부터 신경세포를 보호하는 효과가 있다는 것을 알 수 있었다. The neuroprotective effects of extracts from various parts of peanut sprouts on glutamate-induced neurotoxicity in N18-RE-105 cells were investigated. This study was performed to evaluate the neuroprotective activity of methanolic extracts from the whole (WME), heads (HME), and stems (SME) of peanut sprouts. The neuroprotective effects of these extracts were measured by MTT reduction assay, LDH release assay, phase-contrast microscopy, and flow cytometric analysis on the N18-RE-105 cells. Among these extracts, the HME showed the greatest neuroprotective effects, and was further fractionated with hexane, diethyl ether, ethyl acetate, and water, according to degree of polarity. Out of the fractionated extracts, the diethyl ether layer showed the highest activity on glutamate-induced cytotoxicity in N18-RE-105 cells. The sub-G1 DNA contents of the glutamate-induced severely apoptotic N18-RE-105s were measured by flow cytometric analysis to confirm the HME's anti-apoptotic activity. Interestingly, after incubation with 100 mg/ml of the HME, the proportion of sub-G1 cells of the glutamate-stressed N18-RE-105s had been greatly reduced, from 58.5% to 9.1%. These results imply that HME may have strong potential as a chemotherapeutic agent against neuronal diseases.