1.Exploring Therapeutic Effect of Yuejuwan on Depressed Mice Based on Lipidomics
Zhentao ZHANG ; Dan SU ; Huizhen LI ; Yonggui SONG ; Huanhua XU ; Meixizi LAI ; Zhifu AI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):85-94
ObjectiveTo investigate the effect of Yuejuwan on lipid metabolism in serum, prefrontal cortex and hippocampus of depressed mice based on lipidomics, and to explore the potential pathways for improving lipid metabolism to prevent depression. MethodsSeven-week-old C57BL/6 mice were randomly divided into blank group, model group, Yuejuwan group(3.6 g·kg-1) and fluoxetine group(10 mg·kg-1), and chronic unpredictable mild stress(CUMS) was used to establish the depression model. After 3 weeks of modeling, each administration group was gavaged with the corresponding drug solution according to the dose, and mice in the blank and model groups were given an equal volume of deionised water by gavage, one time/d for 2 weeks. After administration, the antidepressant effect of Yuejuwan was evaluated by neurobehavioral indices such as sucrose preference test, open field test, tail suspension test and forced swimming test. An automatic biochemical analyzer was used to measure contents of total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in mouse serum. Lipidomic analysis of mouse serum, prefrontal cortex and hippocampus was performed based on ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap mass spectrometry(UPLC-LTQ-Orbitrap-MS), and the expression of mammalian target of rapamycin(mTOR), ribosomal protein S6 kinase(S6K), phosphorylation(p)-mTOR, p-S6K in gastric tissues of mice was detected by Western blot. ResultsCompared with the blank group, mice in the model group exhibited significantly reduced sucrose preference rate and center movement time in the open field test(P<0.01), the immobility times in the tail suspension test and forced swimming test were significantly increased(P<0.01), and serum levels of TC, TG, LDL-C, HDL-C, AST and ALT were significantly elevated(P<0.05, P<0.01). Compared with the model group, the Yuejuwan group showed a significant increase in the sucrose preference rate and center movement time in the open field test(P<0.01), the immobility times in the tail suspension test and forced swimming test were significantly reduced(P<0.01), and the serum levels of TC, TG, LDL-C, AST and ALT were significantly decreased(P<0.05, P<0.01). Lipidomic analysis revealed that Yuejuwan had a significant effect on lipid metabolism in serum, prefrontal cortex and hippocampus of depressed mice, and The differential lipid metabolites were mainly enriched in the metabolic pathways of glycerophospholipid metabolism, sphingolipid signaling, and glycosylphosphatidylinositol-anchored protein biosynthesis, among which the glycerophospholipid metabolic pathway was the most significant. Western blot results showed that compared with the blank group, the relative expression levels of p-mTOR/mTOR and p-S6K/S6K in the gastric tissues of mice in the model group were significantly increased(P<0.01). In comparison with the model group, the relative expression levels of p-mTOR/mTOR and p-S6K/S6K in the gastric tissues of mice in the Yuejuwan group were significantly decreased(P<0.01). ConclusionThe intervention of Yuejuwan on lipid metabolism is one of the potential pathways for its antidepressant effect, which may be related to the regulation of mTOR/S6K signaling pathway upstream of lipid metabolism in the gastric tissues.
2.Screening of Antidepressant Active Components from Curcumae Rhizoma and Its Mechanism in Regulating Nrf2/GPX4/GSH Pathway
Yonggui SONG ; Delin DUAN ; Meixizi LAI ; Yali LIU ; Zhifu AI ; Genhua ZHU ; Huanhua XU ; Qin ZHENG ; Ming YANG ; Dan SU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):211-221
ObjectiveTo screen and evaluate the antidepressant compounds of Curcumae Rhizoma, and explore its mechanism of regulating the nuclear factor erythroid 2-related factor 2(Nrf2)/glutathione(GSH) peroxidase 4(GPX4)/GSH pathway from an antioxidant perspective. MethodsThe antioxidant activities in vitro of 11 characteristic components from Curcumae Rhizoma, including curcumol, curgerenone, curdione, curzerene, curcumenol, curcumenone, dehydrocurdione, isocurcumenol, furanodienone, furanodiene and zederone, were detected using 1,1-diphenyl-2-picrylhydrazyl(DPPH) and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt(ABTS) radical scavenging assays. The depression in Drosophila melanogaster was induced by chronic unpredictable mild stress(CUMS), and W1118 wild-type male D. melanogaster were randomly divided into blank group, model group, curcumol group, curgerenone group, curdione group, curzerene group, curcumenol group,curcumenone group, dehydrocurdione group, isocurcumenol group, furanodienone group, furanodiene group, zederone group and fluoxetine group(10 μmol·L-1). The treatment groups received a dose of 0.1 g·L-1 of 11 characteristic components from Curcumae Rhizoma, while the blank and model groups were administered equivalent volumes of solvent. The sucrose preference test, climbing test and forced swimming test were used to evaluate the behavioral indicators of depression in D. melanogaster. Liquid chromatography-mass spectrometry(LC-MS) was used to detect the levels of 5-hydroxytryptamine(5-HT) and dopamine(DA) in the brain of D. melanogaster, and the entropy weight method was used to comprehensively evaluate neurobehavioral and neurotransmitter indicators, resulting in the identification of the antidepressant active components of Curcumae Rhizoma. In addition, a mouse depression model was established by CUMS, and C57BL/6J mice were randomly divided into blank group, model group, low and high dose groups of curzerene(0.5, 1 mg·kg-1), and fluoxetine group(10 mg·kg-1) to confirm the antidepressant effect of the optimal active ingredient by behavioral analysis. Flow cytometry was used to detect the content of reactive oxygen species(ROS) in the hippocampus of mice from each group. Enzyme-linked immunosorbent assay was used to detect the contents of adenosine triphosphate(ATP), superoxide dismutase(SOD), catalase(CAT) and GSH. Transmission electron microscope(TEM) was used to observe the effect of curzerene on the ultrastructure of mitochondria in hippocampal tissue. Western blot was performed to determine the level of Nrf2 protein, and Nrf2 inhibitor(ML385) was used to verify the relationship between the antidepressant effect of curzerene and regulation of Nrf2. Real time fluorescence quantitative polymerase chain reaction(Real-time PCR) was employed to detect the effect of curzerene on the mRNA expression level of GPX. ResultsIn vitro antioxidant experiments showed that curzerene and curgerenone exhibited the most significant ability to scavenge free radicals, and comprehensive evaluation results of entropy weight method indicated that curzerene stood out as the most promising active component. Compared with the blank group, the model group exhibited a significant decrease in sucrose preference coefficient and the number of times entering the open field center(P<0.01), as well as a significant increase in immobility time in the forced swimming and tail suspension tests(P<0.01), and the ROS content in hippocampus significantly elevated(P<0.01), while the ATP content significantly reduced(P<0.01). In the hippocampal neurons of the model group, mitochondrial cristae were disordered, with vacuolation of the inner membrane and severe damage. Nrf2 protein expression level in the model group was significantly decreased(P<0.05), and the antioxidant enzymes SOD, CAT and GSH contents were also significantly reduced(P<0.05, P<0.01), and the gene expression levels of GPX1, GPX4 and GPX7 were significantly decreased(P<0.01). Compared with the model group, the high-dose group of curzerene showed a significant increase in the sucrose preference coefficient and the number of times entering the open field center(P<0.05), as well as a significant decrease in immobility time in the forced swimming and tail suspension tests(P<0.05, P<0.01). The ROS content in the hippocampus of the high-dose group of curzerene was significantly reduced(P<0.01), while the ATP content was significantly increased(P<0.05). The neuronal mitochondrial damage in the hippocampus of the high-dose group of curzerene was alleviated, and the expression level of Nrf2 protein was significantly increased(P<0.05). The Nrf2 inhibitor ML385 reversed the improvement of curzerene on depressive behaviors in CUMS mice. The GSH content in the hippocampal neurons of the high-dose group of curzerene was significantly increased(P<0.01), while there were no significant differences in SOD and CAT contents. The expression level of GPX4 gene in the hippocampal neurons of the high-dose group of curzerene was significantly increased(P<0.05), while there were no significant differences in other GPX genes. ConclusionCurzerene is the best component with antidepressant activity in Curcumae Rhizoma. It may improve mitochondrial dysfunction to exert its antidepressant effect by regulating Nrf2 and its downstream GPX4/GSH pathway rather than CAT or SOD pathways.
3.Effect of Different Fermentation Conditions on Fungal Community and Chemical Composition of Aurantii Fructus
Zhihong YAN ; Xiumei LIU ; Qiuyan GUAN ; Yonggui SONG ; Zhifu AI ; Genhua ZHU ; Yuhui PING ; Ming YANG ; Qin ZHENG ; Huanhua XU ; Dan SU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):254-262
ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus, in order to obtain the optimal fermentation conditions and flora structure, and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2(ITS2) high-throughput sequencing, combined with fungal community diversity analysis and fungal community structure analysis, were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7, 14, 21 d(numbered Y1-Y3 for the former, and numbered F1-F3 for the latter), respectively. At the same time, the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS), combined with principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA) and compound retention time, parent ions, characteristic fragment ions and other information, the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method, while in the non-pressure-shelf natural fermentation method, there was a significant difference with the fermentation process, and at the genus level, the dominant genus of samples Y1, Y2, Y3 and F2 was Aspergillus, while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable, and the microbial community structure was more stable, which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis, including 70 flavonoids, 38 coumarins, 10 alkaloids, 34 organic acids and 3 other compounds. After fermentation, two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition, multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods, mainly including flavonoids, organic acids and coumarins. Comprehensively, the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable, the species richness was high and the overall content of differential compounds was high, which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation, the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds, and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus, as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.
4.Effect of Different Fermentation Conditions on Fungal Community and Chemical Composition of Aurantii Fructus
Zhihong YAN ; Xiumei LIU ; Qiuyan GUAN ; Yonggui SONG ; Zhifu AI ; Genhua ZHU ; Yuhui PING ; Ming YANG ; Qin ZHENG ; Huanhua XU ; Dan SU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(11):254-262
ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus, in order to obtain the optimal fermentation conditions and flora structure, and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2(ITS2) high-throughput sequencing, combined with fungal community diversity analysis and fungal community structure analysis, were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7, 14, 21 d(numbered Y1-Y3 for the former, and numbered F1-F3 for the latter), respectively. At the same time, the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS), combined with principal component analysis(PCA), partial least squares-discriminant analysis(PLS-DA) and compound retention time, parent ions, characteristic fragment ions and other information, the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method, while in the non-pressure-shelf natural fermentation method, there was a significant difference with the fermentation process, and at the genus level, the dominant genus of samples Y1, Y2, Y3 and F2 was Aspergillus, while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable, and the microbial community structure was more stable, which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis, including 70 flavonoids, 38 coumarins, 10 alkaloids, 34 organic acids and 3 other compounds. After fermentation, two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition, multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods, mainly including flavonoids, organic acids and coumarins. Comprehensively, the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable, the species richness was high and the overall content of differential compounds was high, which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation, the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds, and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus, as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.
5.Screening and content determination of differential quality markers in Zingiber officinale mixed and triturated with Schisandra chinensis before and after processing
Pei ZHONG ; Jianglin XUE ; Quan ZHAO ; Chanming LIU ; Xiaojing YAN ; Dan SU ; Yonggui SONG ; Tulin LU ; Wei HUANG
China Pharmacy 2024;35(23):2870-2876
OBJECTIVE To screen and quantitatively analyze differential quality markers (Q-Marker) in Zingiber officinale mixed and triturated with Schisandra chinensis (ZMTS) before and after processing. METHODS HPLC fingerprints of before processing [Z. officinale complicated with S. chinensis (ZWS)] and after processing (ZMTS) (10 batches each) were established. The differences of Q-Markers before and after processing were screened by the chemical pattern recognition method and Q-Marker “five principles”, and the contents were determined. RESULTS A total of 14 common peaks were identified in the fingerprints of ZWS, 22 common peaks were identified in the fingerprints of ZMTS, and 8 components were identified. Differential Q-Marker were screened by chemical pattern recognition and Q-Marker “five principles”, i. e. 6-gingerol, schisandrol A schisandrol B, 8-gingerol, 10-gingerol, schisandrin A, schisandrin B, schizandrin C. The average contents of the 8 differential Q-Markers in ZMTS were 229.46, 244.48, 39.96, 44.12, 61.17, 47.82, 100.11 and 9.70 μg/g, respectively. The average contents of the 4 differential Q-Markers (6-gingerol, schisandrol A, schisandrol B, 8-gingerol) in ZWS were 112.58, 19.01, 26.74 and 5.98 μg/g, respectively. CONCLUSIONS In this study, the differential Q-Markers before and after ZMTS processing are screened. The contents of the Q-Markers in ZMTS after processing are higher than those before processing.
6.Effects of Mosla chinensis seed oil on sleep,olfactory ability,and antioxidant indexes in D.melanogaster
Mengting XU ; Yuchen ZHU ; Dan SU ; Yonggui SONG ; Wenkai ZHANG ; Lei XU ; Qiuting MA ; Zhiyong LIU ; Shaoyong GUO
Acta Laboratorium Animalis Scientia Sinica 2024;32(9):1182-1190
Objective To investigate the effects of MCSO on physiological behavior and antioxidant index in D.melanogaster.Methods One-day-old wild type D.melanogaster was divided into control group,0.25%,0.5%,1%,2%and 4%dose groups,as well as male and female groups.The control group was exposed to the base medium,and each dose group was exposed to the MCSO medium added with 0.25%,0.5%,1%,2%and 4%concentrations,respectively.The optimal dosage concentration and time of administration were investigated by climbing experiment.Then the flies were divided into control group,model group and MCSO group.The model group was established by depriving the flies of sleep through repeated nocturnal light stimulation.Period of drug treatment,appetite test,negative geotaxis ability test,stress test,olfactory memory test,and sleep-wake rhythm detection were used to explore the effects of MCSO on their physiological behavior.The activities of super oxidase dismutase(SOD),catalase(CAT),and malondialdehyde(MDA)were detected by enzyme-linked immunosorbent assay.Results MCSO enhanced the locomotory ability of 30-day-old D.melanogaster(P<0.01),increased the activity of SOD and CAT(P<0.01),and decreased the concentration of MDA(P<0.01).Improve olfactory memory of senile fruit flies.After sleep deprivation,the night sleep time of female Drosophila model group was reduced(P<0.05),and that of male Drosophila model group was reduced(P<0.01).After feeding MCSO,the night sleep time of female drosophila model group was extended(P<0.05),and that of male drosophila model group was extended(P<0.01).Conclusions MCSO had a certain antioxidant effect,prolonging the sleep time and improving the olfactory memory of sleep-deprived Drosophila.
7.Effects of raw and wine-processed Schisandra chinensis on neuro-endocrine-immune network in insomnia mice and its mechanism
Ruiying WANG ; Dan SU ; Huizhen LI ; Yali LIU ; Genhua ZHU ; Ming YANG ; Zhifu AI ; Tao LUO ; Bing XUE ; Yonggui SONG
China Pharmacy 2023;34(5):525-530
OBJECTIVE To investigate the effect of raw and wine-processed Schisandra chinensis on neuro-immune-endocrine network in insomnia mice and its mechanism. METHODS Fifty mice were randomly divided into blank group, model group, diazepam group, raw S. chinensis group and wine-processed S. chinensis group, with 10 mice in each group. Except for blank group, the mice in the other groups were intraperitoneally injected with thyroxine solution to establish mice model of insomnia; at the end of each day’s modeling, the corresponding doses of diazepam,raw and wine-processed S. chinensis were given by gavage. The blank group and model group were given constant volume of normal saline. The general state of the mice was observed and recorded, and the total activity distance and upright times of the mice were detected; the EEG and EMG signals of mice were recorded, and the time ratio of sleep wake time (wake), non-rapid eye movement (NREM) and rapid eye movement (REM) was analyzed; the contents of neurotransmitters [γ-aminobutyric acid (GABA), 5-hydroxytryptamine (5-HT), dopamine (DA), norepinephrine (NE), cortisol (CORT)] in brain suprachiasmatic nucleus (SCN) were detected; and the expressions of tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) were detected; the mRNA expressions of clock gene Bmal1, circadian clock gene Clock and cycle gene Per2 were all detected. RESULTS Compared with the blank group, the mental state of the model group mice was relatively depressed, the amount of food and water increased, the body mass decreased, the hair was rough and shiny, and the circadian rhythm was irregular; the total activity distance and upright times decreased significantly; the time ratio of wake increased significantly, while the time ratios of REM and NREM decreased significantly; the content of 5- HT in brain SCN decreased significantly, while the content of NE, DA and CORT increased significantly; the fluorescence intensity of IL-1β and TNF-α was significantly increased; the relative expression level of Bmal1 and Clock mRNA was significantly increased, while the relative expression level of Per2 mRNA was significantly decreased (P<0.05 or P<0.01). Compared with the model group, the general state of mice in diazepam group, raw S. chinensis group and wine-processed S. chinensis group was improved obviously, and most of the above index levels were significantly reversed (P<0.05 or P<0.01). CONCLUSIONS Raw and wine-processed S. chinensis have a certain therapeutic effect on insomnia mice, the mechanism of which may be related to the regulation of neuro-endocrine-immune system related biological indicators in insomnia mice.
8.Effects of carnosine on ferroptosis and inflammatory responses in STZ⁃induced diabetic mice
Song Zhang ; Xueqi Liu ; Ling Jiang ; Yonggui Wu
Acta Universitatis Medicinalis Anhui 2023;58(8):1322-1328
Objective :
To investigate the effects of carnosine (CAR) on streptozotocin (STZ) induced renal feroptosis and inflammation in diabetic mice .
Methods :
Type 1 diabetes mice model were induced by STZ , and normal C57 mice were used as normal control group . The C57 mice were divided into 5 groups (6 - 8 mice in each group) : normal control group (NC) , normal control + carnosine group (NC + CAR) , STZ model group (STZ) ,STZ model + carnosine group (STZ + CAR) , STZ model + ferroptosis inhibitor group (STZ + Fer⁃1) . After feeding the mice for 16 weeks , serum samples were collected to detect blood creatinine ( CRE) and urea nitrogen ( BUN) levels . The urine of mice was collected to detect the 24 - hour urinary albumin level . HE staining and PAS staining were performed to observe the degree of renal pathological injury . Real⁃time PCR was used to detect the expression of interleukin (IL) Ⅳ1β , IL⁃6 , monocyte chemotactic protein 1(MCP⁃1) and tumor necrosis factor⁃α (TNF⁃α ) in mouse kidney tissue . The expression of reactive oxygen species(ROS) in mouse kidney was detected by immunofluorescence . Morphology of renal mitochondria was observed by transmission electron microscopy . The protein expression levels of glutathione peroxidase 4(GPX 4) and long⁃chain lipoacyl⁃CoA synthetase 4( ACSL4) , which are ferroptosis indexes , were detected by Western blot . The contents of malondialdehyde ( MDA) , glutathione ( GSH) and Fe2 + in mouse kidney tissue were determined .
Results :
Compared with NC group , CRE and BUN levels in STZ group increased (P < 0. 001) ; and ompared with STZ group , these indexes decreased in STZ + CAR group (P < 0. 001 , P < 0. 01) . Renal histopathological examination showed that compared with NC group , renal tubule dilatation , inflammatory cell infiltration and glycogen deposition significantly increased in STZ group ; and compared with STZ group , tubule dilatation , inflammatory cell infiltration and glycogen deposition decreased in STZ + CAR group . Electron microscope results showed that the renal mitochondria in STZ group were swollen , membrane density increased , mitochondrial ridges decreased or absent , renal tubule dilation was improved significantly in the STZ + CAR group and STZ + Fer⁃1 group , and inflammatory cell infiltration was reduced . Real⁃time PCR test results showed that compared with NC group , mRNA expression levels of inflammatory factor (IL⁃1β , IL⁃6 , MCP⁃1 and TNF⁃α ) increased in STZ group (P < 0. 001 or P < 0. 01) ; and mRNA expressions of IL⁃1β , IL⁃6 , MCP⁃1 and TNF⁃α were decreased in STZ + CAR group compared with STZ group (P < 0. 01 or P < 0. 05) . Immunofluorescence results showed that compared with NC group , ROS level in kidney tissue of mice in STZ group increased (P < 0. 001) ; and compared with STZ group , the expression of ROS in kidney tissue of STZ + CAR group decreased while ROS expression in STZ + CAR group decreased ( P < 0. 01) . Compared with NC group , GPX4 expression and GSH content in kidney of STZ group decreased ( P < 0. 001) , and ACSL4 protein expression and MDA and Fe2 + contents increased (P < 0. 01 or P < 0. 001) , GPX4 expression and GSH content increased (P < 0. 01) , ACSL4 protein expression and MDA and Fe2 + content decreased in STZ + CAR group (P < 0. 01 or P < 0. 001) .
Conclusion
CAR inhibits ferroptosis and inflammation in the kidney in diabetic mice induced by STZ ,
and improved renal pathological injury in diabetic mice .
9.Full spectrum analysis of chemical constituents of Sargassum fusiforme and its in vitro anti-neuroinflammatory activity
Hong PENG ; Pinzhe HUANG ; Yonggui SONG ; Huanhua XU ; Mingyue ZHOU ; Genhua ZHU ; Ming YANG ; Zhifu AI ; Dan SU
China Pharmacy 2022;33(7):800-807
OBJECTIVE To study the composition of chemical constituents of Sargassum fusiforme and its in vitro anti- neuroinflammatory activity ,and to provide reference for its development and utilization and the study of pharmacodynamic substances. METHODS UHPLC-QTOF-MS/MS analysis method and GC-MS/MS method were used to analyze the chemical constituents of S. fusiforme . The lipopolysaccharide (1 μg/mL)was adopted to establish the inflammatory model of neuromicroglia BV2. Using paroxetine (5 μg/mL)as positive control ,CCK-8 assay was used to detect the effects of the extracts of S. fusiforme (20,40,60,80,100 μg/mL)on the activity and morphology of neuromicroglia BV 2. The effects of the extracts of S. fusiforme (40,60,80 μg/mL)on the contents of tumor necrosis factor α(TNF-α)and interleukin- 6(IL-6)in cell supernatant were detected by ELISA. RESULTS A total of 103 non-volatile constituents were identified by UHPLC-QTOF-MS/MS ,and 60 volatile constituents were obtained by GC-MS/MS. The extracts of S. fusiforme (40,60,80 μ g/mL) could significantly reduce the abnormally increased activation of neuromicroglia BV 2 and the contents of TNF-α and IL-6 due to lipopolysaccharide (P<0.05 or P<0.01). CONCLUSIONS The study establish the full spectrum of chemical constituents of S. fusiforme ,and it is confirmed that fusiforme has certain in vitro anti-neuroinflammatory activity.
10.Compatibility Advantage of Scutellariae Radix-Coptidis Rhizoma for Anti-neuroinflammation and Its Potential Targets for Regulating TLR4/MyD88/NF-κB Signaling Pathway
Hongjie ZHANG ; Dan SU ; Genhua ZHU ; Yonggui SONG ; Bugao ZHOU ; Shanshan LI ; Changhua ZHANG ; Zhifu AI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(22):58-67
ObjectiveTo explore the compatibility advantage of Scutellariae Radix-Coptidis Rhizoma in the prevention and treatment of neuroinflammation, and to elucidate the action characteristics and mechanism of the compatibility advantage based on Toll like receptor (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor kappaB (NF-κB) pathway. MethodRepresentative mouse microglia cells (BV2) in vitro were selected and divided into 8 groups: control group, model group, Scutellariae Radix-Coptidis Rhizoma group, Piracetam group, Scutellariae Radix group and Coptidis Rhizoma group. The BV2 cell inflammatory model was established by lipopolysaccharide (LPS), and the cell activity was detected by cell counting kit-8 (CCK-8). Cell morphology was observed under bright field. The production and release of pro-inflammatory factors in BV2 cells were determined by enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assay, and the mRNA expressions of TLR4, MyD88 and NF-κB were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). The nuclear translocation of NF-κB p65 was detected by immunofluorescence, and TLR4 signal transduction inhibitor (CLI-095) and NF-κB inhibitor (PDTC) were used to confirm the anti-neuroinflammation targets of Scutellariae Radix-Coptidis Rhizoma. ResultCompared with the conditions in the control group, most cells in LPS-induced model group were activated, and the contents of IL-6, TNF-α and IL-1β in culture medium and cells and the mRNA expressions of TLR4, MyD88, NF-κB p50 and NF-κB p65 were increased (P<0.01), with obvious nuclear entry of NF-κB p65. Compared with the conditions in the model group, BV2 cell morphology was mostly recovered after pretreatment in Scutellariae Radix-Coptidis Rhizoma and Piracetam groups, and the levels of IL-6, TNF-α and IL-1β and the mRNA expressions of TLR4, MyD88, NF-κB p50 and NF-κB p65 were decreased (P<0.05, P<0.01), with NF-κB p65 mostly observed in cytoplasm. Compared with the conditions in the model group, cell morphology was slightly recovered in Scutellariae Radix group and Coptidis Rhizoma group, and the levels of pro-inflammatory factors and mRNA expressions of TLR4, MyD88, NF-κB p50 and NF-κB p65 were reduced. In terms of inhibitory effect on pro-inflammatory factors, Scutellariae Radix group and Coptidis Rhizoma group were lower than Scutellariae Radix-Coptidis Rhizoma group (P<0.05). Compared with the model group, the "Scutellariae Radix-Coptidis Rhizoma+CLI-095" group and "Scutellariae Radix-Coptidis Rhizoma+PDTC" group had lowered mRNA expressions of TLR4, MyD88, NF-κB p50 and NF-κB p65 (P<0.05, P<0.01), and the transfer of NF-κB p65 into nucleus was obviously inhibited. ConclusionThe anti-neuroinflammation effect of Scutellariae Radix-Coptidis Rhizoma was significantly better than Scutellariae Radix or Coptidis Rhizom alone, and the anti-neuroinflammation advantage was closely related to the inhibition of activation of TLR4/MyD88/NF-κB signaling pathway in microglial cells. It was confirmed that TLR4, MyD88 and NF-κB were potential targets for Scutellariae Radix-Coptidis Rhizoma to exert the compatibility advantage.


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