Cognitive Enhancing Activity of Lysimachia christinae Extract on Scopolamineinduced Memory Impairment in Mice

Alzheimer’s disease (AD) is characterized by the gradual loss of memory and learning abilities, representing a form of dementia. Lysimachia christinae (LC), a traditional medicinal plant native to China, Japan, and Korea, is esteemed for its reputed cholagogic, hepatoprotective, and antihyperlipidemic properties.Previous research from our team has demonstrated the neuroprotective effects of LC extract in HT22 cells exposed to glutamate insult. This study seeks to further explore LC’s potential in enhancing cognitive function, utilizing a mouse model with scopolamine-induced memory impairment. The Morris water maze test assessed spatial memory enhancement, while the passive avoidance test evaluated its effect on learning memory. LC was extracted using methanol and an ultrasonic extraction device. Subsequently, the LC extract was administered to ICR mice subjected to scopolamine insult at concentrations of 100, 200, and 300 mg/kg, respectively. Notably, the LC extract significantly improved memory impairment induced by scopolamine and inhibited acetylcholinesterase activity. These results suggest that the LC extract ameliorated memory impairment by enhancing the acetylcholine esterase signaling pathway. Based on these findings, we propose that the LC extract shows promise as a potential candidate for the development of new nutraceuticals aimed at improving memoryimpairment.

Simultaneous Quantification of Eight Compounds of Lonicera japonica by HPLC-DAD

Lonicera japonica are recognized in traditional oriental medicine for their notable antiviral, antiinflammatory, and antibacterial attributes. According to the findings from the previous study, it demonstrates antioxidative and neuroprotective activity. Analytical methods for the simultaneous quantification of eight compounds isolated from L. japonica, loganin (1), secoxyloganin (2), caffeic acid (3) rutin (4), hyperoside (5), lonicerin (6), quercetin (7) and luteolin (8) were established through the utilization of HPLC-DAD. This HPLC analysis was conducted using a Luna C18 column (5 μm, 4.6 mm × 150 mm) at 25 o C. The mobile phase, comprising 0.1% trifluoroacetic acid and acetonitrile, was run at a flow rate of 1 mL/min. Validation of the method included assessments for linearity, precision and accuracy. The calibration curve displayed exceptional performance, boasting an r² value surpassing 0.9989. Limits of detection (LOD) ranged from 1.11 to 3.18 mg/ mL, while limits of quantification (LOQ) spanned from 3.33 to 9.54 mg/mL. In the precision test, both intra- and inter-day assessments revealed minimal relative standard deviations (RSD) values, consistently below 2.91% and 3.48%, respectively. Accuracy test outcomes fell within the 98.22% to 103.47% range, with RSD values consistently under 2.68%. These findings affirm the HPLC-DAD method’s high reliability and accuracy in the quantitative analysis of eight compounds in L. japonica extract, rendering it well-suited for quality control purposes.

Anti-influenza Compounds Isolated from Descurainia sophia Seeds

Descurainia sophia seeds methanol extract showed significant anti-influenza activity and we tried to isolate anti-influenza compounds from the D. sophia extract. D. sophia seeds were extracted with 80% methanol and fractionated with n-hexane, ethyl acetate, CHCl 3 and n-butanol. The anti-influenza activity of each fraction was assessed using sulforhodamine B (SRB) method in A549 cells, human-derived lung cancer cells. The ethyl acetate and CHCl 3 fractions showed the most potent anti-influenza activity. Seven compounds were isolated from CHCl 3 fraction and identified 1-decanol (1), 2-(3,4-dihydroxy-2-methylenebutoxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (2), daucosterol (3), isorhamnetin (4), quercetin (5), sinapic acid (6), and helveticoside (7) by spectroscopic data such as UV, IR, 1 H-NMR, 1 C-NMR and mass spectroscopy. Anti-influenza activities of isolated compounds were evaluated using SRB method in A549 cells. Compounds 3, 4 and 7 had significant antiinfluenza activity in a dose-dependent manner.

Therapeutic Efficacy of Methanol Extract of Bidens tripartita in HT22 Cells by Neuroprotective Effect

Oxidative stress brings about apoptosis through various mechanisms. In particular, oxidative stress in neuronal cells can causes a variety of brain diseases. This study was conducted to investigate the effect of Bidens tripartita on oxidative stress in neuronal cells. B. tripartita has traditionally been used in Russia as a medicine for diseases such as rhinitis, angina and colitis. Over-production of glutamate induces oxidative stress. When the oxidative stress occurs in the cells, reactive oxygen species (ROS) and Ca 2+ increase. In addition, the abrupt decline of mitochondrial membrane potential and the decrease of glutathione related enzymes such as glutathione reductase (GR) and glutathione peroxidase (GPx) are also observed. The samples used in the experiment showed cytoprotective effect in the MTT assay. It also lowered the ROS and Ca 2+ level, and increased degree of mitochondrial membrane potential, GR and GPx. As a result, B. tripartita had a positive effect against oxidative stress. Thus, it is expected to have potential for treatment and prevention of degenerative brain diseases such as Alzheimer’s disease.

Neuroprotective Compounds Isolated from Lysimachia christinae

We previously reported that dried Lysimachia christinae whole plant extract exerted significant neuroprotective activity. In this study, we tried to isolate neuroprotective compounds of L. christinae. We evaluated the neuroprotective activity of the four fractions (hexane, chloroform, ethyl acetate, and n-butanol fractions) of methanol extract. Among them, ethyl acetate and n-butanol fractions showed most potent neuroprotective activity against glutamate excitotoxicity. Nine compounds were isolated from ethyl acetate and n-butanol fractions of L. christinae extract and identified as cynaroside (1), (3,4,5,6-tetrahydroxytetrahydro-2Hpyran-2-yl)methyl-3-hydroxy-2-octyldopentaconta-23,33-dienoate (2), androst-16-ene-3,6-diol (3), 2-hydroxy-24-propoxytetracos-4-enoic acid (4), 2-hydroxy-24-methoxytetracos-4-enoic acid (5), 12-(stearoyloxy)octadec-9-enoic acid (6), β-sitosterol (7), (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-yl palmitate (8) and (1S,2S,3R,4R)-4-(((2S,3R,4R,5R,6S)-2-(((2R,3R,4S,5R,6R)-2-(3,4-dimethoxyphenethoxy)-3,5-dihydroxy-6-(hydroxymethyl) tetrahydro-2H-pyran-4-yl)oxy)-4,5-dihydroxy-6-methyltetrahydro-2H-pyran-3-yl)oxy)cyclohexane-1,2,3-triol (9) by spectroscopic data such as UV, IR, NMR, Mass spectroscopy. Their neuroprotective activity was evaluated by MTT assay. Cynaroside (1) and androst-16-ene-3,6-diol (3) had significant neuroprotective activity against glutamate-injured HT22 cells. The neuroprotective efficacy of cynaroside (1) and androst-16-ene-3,6-diol (3) was related to their anti-oxidative activity.

Simultaneous Determination of Five Compounds in Descurainia sophia by HPLC-DAD

Descurainia sophia was commonly used for treating of cough and asthma in Asian country such as Korea, China and Japan. According to our previous study, it also had anti-influenza activity and anti-influenza compounds were isolated and identified. In this study, simultaneous determination analytical method of isolated five compounds such as daucosterol (1), helveticoside (2), isorhamnetin (3), quercetin (4), and sinapic acid (5) was established by using HPLC-DAD. The HPLC analysis was carried out using a Dionex C 18 column (5 µm particle size, 120 Å pore size, 4.6 mm × 150 mm dimensions) at a constant temperature of 25 o C. The mobile phase consisted of a blend of 0.1% trifluoroacetic acid (TFA) and acetonitrile and delivered at a flow rate of 1 mL/min. Method validation was conducted to assess its reliability and accuracy. The calibration curve exhibited excellent linearity, with an R² value exceeding 0.9994, demonstrating the method’s capability to accurately quantify the compounds across a range of concentrations. Limit of Detection (LOD) ranged from 0.93 to 3.19 µg/ mL, and limit of quantification (LOQ) spanned from 2.79 to 9.57 µg/mL, respectively. The relative standard deviations (RSD) values of precision test, intra- and inter- day, were less than 1.28 % and 1.14%. The accuracy test results ranged from 100.81% to 105.31% and RSD values were less than 1.26 %. These results showed that the HPLC-DAD method was very reliable and accurate for the quantity analysis of eight compounds in D. sophia extract for quality control.

Simultaneous Determination of Eight Compounds in Lysimachia christinae by HPLC-DAD

Lysimachia christinae Hance was commonly used in Oriental medicine for treating the hepatitis virus, cholecystitis and cholagogic efficiency. According to the previous study, it possesses high antioxidant and anti-inflammatory activity. Simultaneous determination analytical method of isolated eight compounds, cynaroside (1), 2-(3,4-dimethoxyphenyl) ethyl O-α-L-arabinopyranosyl-(1→2)-O-[6-deoxy-α-L-mannopyranosyl-(1→3)] β-Dglucopyranoside (2), stearylester ricinoleic acid (3), (E)-4-(3,4-dimethoxyphenyl) but-3-en-1-yl palmitate (4), 2-hydroxy-24-methoxy-4-tetracosenoic acid (5), 2-hydroxy-24-propoxy-4-tetracosenoic acid (6), β-sitosterol (7), and androst-16-ene-3,6-diol (8) were established by using HPLC-DAD. This HPLC analysis was detected on a Dionex C18 column (5 ㎛, 120 Å, 4.6 mm × 150 mm) at 25 o C. The mobile phase consisted of 0.1% trifluoroacetic acid and acetonitrile at a flow rate of 1 mL/min. Validation of the method was assessed by linearity, precision and accuracy test. Calibration curve was good at r 2 > 0.9998. Limits of detection (LOD) ranged from 0.19 to 8.18 g/ml and Limits of quantification (LOQ) ranged from 0.19 to 24.80 g/ml. The relative standard deviations (RSD) values of precision test, intra- and inter- day, were less than 0.99% and 1.0%. The accuracy test results ranged from 98.81% to 106.49% and RSD values were less than 0.95%. These results showed that the HPLC-DAD method was very reliable and accurate for the quantity analysis of eight compounds in L. christinae extract for quality control.

Cognitive-Enhancing Effect of Dianthus superbus var. Longicalycinus on Scopolamine-Induced Memory Impairment in Mice

Dianthus superbus (D. superbus) is a traditional crude drug used for the treatment of urethritis, carbuncles and carcinomas. The objective of this study was to confirm the cognitive enhancing effect of D. superbus in memory impairment induced mice and to elucidate the possible potential mechanism. Effect of D. superbus on scopolamine induced memory impairment on mice was evaluated using the Morris water maze and passive avoidance tests. We also investigated acetylcholinesterase (AChE) activity and brain-derived neurotropic factor (BDNF) expression in scopolamine-induced mice. HPLC-DAD analysis was performed to identify active compounds in D. superbus. The results revealed that D. superbus attenuated the learning and memory impairment induced by scopolamine. D. superbus also inhibited AChE levels in the hippocampi of the scopolamine-injected mice. Moreover, D. superbus increased BDNF expression in the hippocampus. Eight compounds were identified using HPLC-DAD analysis. The content of 4-hydroxyphenyl acetic acid was higher than contents of other compounds. These results indicated that D. superbus improved memory functioning accompanied by inhibition of AChE and upregulation of BDNF, suggesting that D. superbus may be a useful therapeutic agent for the prevention or treatment of Alzheimer's disease.

Simultaneous Determination of 11 Marker Compounds in Gumiganghwal-tang by HPLC-DAD and LC-MS

Gumiganghwal-tang has been used for the treatment of common cold for a long-time. We developed an accurate and sensitive high performance liquid chromatography-diode array detection (HPLC-DAD) and electrospray ionization mass spectrometry method for the simultaneous determination of ferulic acid, baicalin, bergapten, methyl eugenol, glycyrrhizin, oxypeucedanin, wogonin, nodakenin, atractylenolide III, imperatorin, and atractylenolide I in Gumiganghwal-tang samples. The analytes were separated on a Shiseido C18 column (5 µm, 4.6 mm I.D. × 250 mm) with gradient elution with acetonitrile and 0.1% trifluoroacetic acid. Eleven compounds were quantitatively determined by HPLC-DAD and identified by LC-MS data. We also validated this method. The calibration curves of all the compounds showed good linear regression. The limits of detection and the limits of quantification ranged from 0.04 to 0.63 and from 0.12 to 1.92 µg/mL, respectively. The relative standard deviation values of intra- and inter-days of this method represented less than 2.9%. The recoveries were found to be in the range of 90.06 – 107.66%. The developed method has been successfully applied to the analysis of Gumiganghwal-tang samples. The established HPLC method could be used to quality control of Gumiganghwal-tang.

Neuroprotective Effect of Steamed and Fermented Codonopsis lanceolata

Codonopsis lanceolata has been used as an herbal medicine for several lung infl ammatory diseases, such as asthma, tonsillitis, and pharyngitis. Previously, we showed the neuroprotective effect of steamed and fermented C. lanceolata (SFC) in vitro and in vivo. In the current study, the treatment of HT22 cells with SFC decreased glutamate-induced cell death, suggesting that SFC protected HT22 cells from glutamate-induced cytotoxicity. Based on these, we sought to elucidate the mechanisms of the neuroprotective effect of SFC by measuring the oxidative stress parameters and the expression of Bax and caspase-3 in HT22 cells. SFC reduced contents of ROS, Ca2+ and NO. Moreover, SFC restored contents of glutathione and glutathione reductase as well as inhibited Bax and caspase-3 activity in HT22 cells. These results indicate that steamed and fermented C. lanceolata (SFC) extract protected HT22 cells by anti-oxidative effect and inhibition of the expression of Bax and caspase-3.