1.Establishing the SREBP Pathway in the Oocytes Xenopus Laevis and Stuyding the Effects of the Cucurmin
Like WO ; Chunlei FANG ; Xingde WO
Journal of Zhejiang Chinese Medical University 2006;0(03):-
[Aim] The expressed system of sterol regulation by SCAP/SREBP Pathway was established in the oocyte of the Xenopus laevis and was applied to investigate the effect of cucurmin on the SREBP pathway.[Methods] The plasmid of PLXRN-4SRE-fpa was restructed and injected into the nucleus of Xenopus Oocytes in phase V or VI. The oocyte was incubated with different concentrations of the curcumin. After three days the fluorescence intensity on the membrane of the Oocytes was measured by Fluoroanalyzer.[Results] The fluoresecence intensity on the membrane of the oocyte increased with increasing of the concentration of the curcumin. The trail group which have all regulation elements of SCAP/SREBP pathway can express more fluorscence protein than the control group. [Conclusion] The experiment provides that the curcumin can upgrade the expression of low density lipoprotein receptor by SCAP/SREBP regulation pathway. The model can be applied for quick screening of the scap ligand drugs and inverstigating the mechanism of the drugs.
2.Changes in cerebral vaso- active substances in patients undergoing intracranial aneurysm clipping under propofol-fentanyl anesthesia
Fang WO ; Shuzhen ZHANG ; Enzhen WANG
Chinese Journal of Anesthesiology 1997;0(11):-
90 bpm and nicardipine 10?g?kg-1 when MAP increased by 20% of the baseline value. Radial artery was cannulated for continuous BP monitoring and blood sampling. MAP, ECG, HR, SpO2 , PET CO2 and body temperature were continuously monitored during anesthesia. Blood samples were taken before induction of anesthesia (T1), when dura was cut (T2), when aneurysm was clipped (T3 ) and 30 min after clipping (T4 ) for determination of plasma concentrations of AT- Ⅱ , ET and CGRP by radioimmuno-assay.Results Plasma concentration of AT- Ⅱ and HR did not change significantly through out the study. Plasma CGRP concentration and MAP decreased significantly during operation at T2-4 as compared to the baseline values (T1 ) . Plasma ET concentration increased significantly at clipping (T3 ) as compared to that at T2. Conclusion The action of vaso-constrictors (AT- Ⅱ , ET) predominates over that of vaso-dilator during operation. It is important to prevent acute cerebral vasospasm during intracranial aneurysm clipping.
3.Effect of Shenmai Injection on angiogenesis in tumor tissue
Chengxian GAO ; Zhishan DING ; Lihui YIN ; Sanhua FANG ; Wei YUAN ; Xingde WO ;
Chinese Traditional Patent Medicine 1992;0(09):-
Objective: To investigate the effect of Shenmai Injection (Radix Ginseng Rubra, Radix Ophiopogonis) on angiogenesis and the expression of proliferating cell nuclear antigen (PCNA) in tumor tissue, and the mechanism of it in the treatment of tumor. Methods: The mouse tumor model was used to investigate the effect of Shenmai Injection on tumor growth on the whole. The expression of von Willebrand factor (vWF) and PCNA in tumor tissue was studied by means of immunohistochemistry. Results: Shenmai inhibited the tumor growth and reduced microvessel density and the expression of PCNA in tumor tissue. Conclusion:Antiangiogenesis is one of the mechanisms of Shenmai Injection in treatment of tumor.
4.Real-time assessments of liver quality in cold ischemia injury based on the technique of laser scanning confocal microscope
Xu FANG ; Zhongyi JIANG ; Lei LI ; Qi WO ; Lin ZHONG
Chinese Journal of Organ Transplantation 2019;40(5):303-307
Objective To assess the quality of donor liver allografts by employing laser scanning confocal microscope (LSCM ) and clinical liver function tests .Methods Sprague-Dawley rats were used for establishing cold ischemia models of liver allografts .According to different timepoints of cold ischemia ,four groups of CIT1h ,CIT6h ,CIT12h and CIT24h were designated .At the end of cold ischemia time (CIT) of each group , perfusion and preservation fluids were collected and fluoresceins perfused . After LSCM examinations ,tissue samples were harvested for HE examination .Finally a comparison was made between LSCM results and hematoxylin & eosin (HE) examinations .Also some relevant clinical parameters were detected in preserving and flushing fluids .Results Both LSCM examination and pathological examination indicated that the quality of liver allografts decreased significantly with the elapsing of time . Only the difference of LDH was statistically significant (P<0 .001) .Conclusions LSCM may be used for evaluating the ex vivo qualities of liver allografts .Simple handling and time efficiency re great advantages of LSCM .As compared with alanine transaminase (ALT ) and aspartate transaminase (AST ) ,LDH is a better indicator reflecting the quality of liver allografts .
5.Effects of propofol sedation on BDNF-TrkB/p75 signal and cognitive function in rat hippocampus
Wen-Juan YU ; Min ZHU ; Yan WO ; Yi-Min YU ; Yan LI ; Hong-Wei FANG ; Hao ZHU
Journal of Shanghai Jiaotong University(Medical Science) 2018;38(6):594-597
Objective·To detect the effects of propofol sedation on cognitive function in rats and its mechanism. Methods?·?Forty-eight SD rats were randomly divided into three groups, i.e. control group, 100?mg/kg group and 300?mg/kg group. Rats were administrated intraperitoneally with propofol (10?mg/mL, 100?mg/kg or 300?mg/kg). The mRNA levels of brain derived neurotropic factor (BDNF)-TrkB/p75 signal molecules in rat hippocampus were evaluated by realtime PCR 45 min after propofol treatment. Learning and memory ability was examined by inhibitory avoidance (IA) test after propofol treatment. Results?·?The mRNA levels of BDNF in the hippocampal tissue were (1.20±0.13) fold (P=0.002) and (88±12) % (P=0.044) of that in control group, respectively, in 100?mg/kg group and 300?mg/kg group after injection of propofol. The mRNA levels of TrkB were (1.01±0.11) fold ( P=0.982) and (86±11) % (P=0.018) of that in control group, respectively, in 100?mg/kg group and 300?mg/kg group. The mRNA levels of p75 were (1.02±0.10) fold (P=0.778) and (1.59±0.18) fold (P=0.000) of that in control group, respectively, in 100?mg/kg group and 300?mg/kg group. There was no significant difference of the 24 h IA memory retention latency between 100?mg/kg group and control group. The 24 h IA memory retention latency in 300?mg/kg group was significantly decreased compared with control group (P=0.028) and 100?mg/kg group (P=0.020). Conclusion?·?Propofol dose-dependently regulates the expression of BDNF-TrkB/p75 signal molecules, and high dose propofol may reduce cognitive function via BDNF-TrkB/p75 signal.
6.Exploring the mechanism of acute liver injury induced by Cortex dictamni based on network pharmacology and in vivo experiments
Wenkai HU ; Xiaomin XU ; Xin JIANG ; Jiameixue WO ; Suxia JIA ; Shumin LIU ; Fang LU
China Pharmacist 2024;27(7):1089-1098
Objective To explore the mechanism of acute liver injury induced by Cortex dictamni through network pharmacology and in vivo experiment in animal.Methods The chemical constituents and targets of Cortex dictamni were retrieved from TCMSP,TCMIP and SwissTargetPrediction databases,and the related targets of liver injury diseases were identified through GeneCards and CTD databases.The protein interaction network of the intersection targets was analyzed by STRING database and the core targets were selected.The GO function and KEGG pathway enrichment analysis were completed by DAVID database,and the multi-level association network diagram of"drug-component-target"was constructed by Cytoscape software.In the animal study,Cortex dictamni was administered to mice at a dosage of 92.7 g/(kg·d)via intragastric administration,and the biological samples were collected after 7 days.The pathological changes of liver were observed by hematoxylin-eosin(HE),Masson and Oil Red O staining.The expression levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),and lactate dehydrogenase(LDH)in serum,as well as malondialdehyde(MDA),superoxide dismutase(SOD),tumor necrosis factor-α(TNF-α),and interleukin(IL)-1β in liver tissues,were quantified using enzyme-linked immunosorbent assay(ELISA).The expressions of protein kinase B1(AKT1),IL-6,TNF-α,tumor protein p53(TP53),cystatin 3(CASP3),and IL-1β mRNA in liver tissues were determined using real-time quantitative reverse transcription PCR(qRT-PCR).Molecular docking was employed to verify the binding affinity of potentially toxic components to their respective targets.Results A total of 14 chemical constituents,244 predicted targets and 202 intersection targets with liver injury were obtained.The GO biological process analysis mainly involved positive regulation of gene expression,negative regulation of apoptosis process.KEGG pathway enrichment analysis mainly included cancer pathway and PI3K-Akt,TNF,IL-17 signaling pathways.The pathological sections revealed severe hemorrhage,a considerable amount of hepatocyte necrosis,nuclear fragmentation or dissolution in the liver tissues of mouse with HE staining after the administration of Cortex dictamni.Masson staining showed evident fibrosis in the liver tissues,while Oil Red O staining indicated a substantial production of lipid droplets.Compared with the control group,the ELISA results demonstrated a significant increase in serum AST,ALT,ALP,LDH levels,as well as hepatic MDA,TNF-α,and IL-1β levels(P<0.05),and a decrease in hepatic SOD levels(P<0.05)in the treated group.The qRT-PCR results indicated a significant elevation in the expression levels of relevant mRNAs in the liver tissues of the treated mice(P<0.05).Molecular docking showed that the potentially toxic components of obacunone,dictamnine and fraxinellon had good binding affinity to AKT1,IL-6,TNF-α,TP53,CASP3 and IL-1β.Conclusion Obacunone,dictamnine,fraxinellon,and limonin might be the potential toxic components of acute liver injury induced by Cortex dictamni in mice.Cortex dictamni could act on the liver by changing the expressions of AKT1,IL-6,TNF-α,TP53,CASP3,IL-1β and other proteins,affecting energy metabolism,cell differentiation,inflammation,oxidative stress and immunity,leading to liver injury.
7.Research progress on chemical constituents,pharmacological effects and toxicity of Dictamni cortex
Wenkai HU ; Xiaomin XU ; Xin JIANG ; Jiameixue WO ; Suxia JIA ; Shumin LIU ; Fang LU
China Pharmacist 2024;28(10):252-267
Dictamni cortex is the root bark of Rutaceae plants.It is the main medicinal part and the key drug of 'Zhuhuang Fengbi'.It has the effects of clearing heat and detoxifying,dispelling wind and drying dampness,and relieving itching.Dictamni cortex mainly contains 228 chemical components such as alkaloids,sesquiterpenes,limonoids,fatty acids,volatile oils,flavonoids,steroids,etw.Its pharmacological activities in vivo and in vitro include antibacterial activity,anti-inflammatory activity,hepatoprotective activity,cardiovascular protection activity,insecticidal activity,anticancer activity,anti-allergic activity,and improvement of gastrointestinal activity.It has been reported that Dictamni cortex also has potential hepatotoxicity,among which dictamnine,fraxinellone and limonin compounds are potential hepatotoxic components.In this paper,the chemical constituents,pharmacological effects and toxicity of Dictamni cortex are reviewed by consulting domestic and foreign literature,to provide theoretical support for the clinical rational application and related product development of Dictamni cortex.