1.Effects and mechanism of total alkaloids of Corydalis Rhizoma on the regulation of cuproptosis in rats with diabetic cardiomyopathy
Jun LI ; Yazhi QI ; Ya TANG ; Rui CAO ; Qiang XU ; Yusheng HAN
China Pharmacy 2025;36(7):801-806
OBJECTIVE To investigate the effects and mechanism of total alkaloids of Corydalis Rhizoma (TAC) on the regulation of cuproptosis in rats with diabetic cardiomyopathy (DCM) based on silence information regulator 1(Sirt1)/tumor protein 53(P53)signaling pathway. METHODS DCM rat model was induced by high-fat and high-sugar diet and intraperitoneal injection of streptozotocin. Thirty-two model rats were randomly divided into model group, TAC low-dose, medium-dose and high-dose groups (7, 10.5, 14 mg/kg), with 8 rats in each group. An additional 8 rats were assigned to normal control group. Related drugs or normal saline were administered intragastrically in each group, once a day, for 4 weeks. After the last medication, the fasting blood glucose (FBG) levels of the rats were measured. The levels of myocardial creatine kinase (CK), creatine kinase isoenzyme (CK-MB), and lactate dehydrogenase (LDH) in serum and myocardial tissue of rats were all detected. The pathological morphology, fibrosis degree, and Cu2+ deposition of myocardial tissue in rats were observed. The levels of Cu2+ and glutathione (GSH) in myocardial tissue, the expressions of Sirt1/P53 signaling pathway-related proteins [Sirt1, P53, solute carrier family 7 membrane 11 (SLC7A11)], and iron-sulfur cluster-related proteins [ferredoxin 1 (FDX1), lipoic acid synthetase (LIAS), aconitase 2 (ACO2), NADH-ubiquinone oxidoreductase core subunit S8 (NDUFS8), dihydrolipoamide acetyltransferase (DLAT), dihydrolipoamide succinyltransferase (DLST)], and heat shock protein 70 (HSP70) were all determined. RESULTS Compared with normal control group, the model group exhibited significantly elevated levels of FBG, CK, CK-MB and LDH in both serum and myocardial tissue, as well as increased 2+ levels of Cu in myocardial tissue and the expression of P53 and HSP70 proteins (P<0.05); the level of GSH and the expression levels of Sirt1, SLC7A11, FDX1, LIAS, ACO2, NDUFS8, DLAT, and DLST proteins in myocardial tissue were all significantly decreased (P<0.05); the myocardial tissue exhibited severe pathological damage, with numerous inflammatory cell infiltrations and significant fibrosis, as well as increased deposition of Cu2+. Compared with model group, most of the above quantitative indicators in rats were significantly reversed in TAC groups (P<0.05); the pathological damage to the myocardial tissue was alleviated, with reduced fibrosis and Cu2+ deposition. CONCLUSIONS TAC can ameliorate DCM in rats, and its mechanism of action may be related to activating the activity of the Sirt1/P53 signaling pathway, promoting the chelation of GSH with Cu2+, and inhibiting cuproptosis of cardiomyocyte.
2.Effect of electroacupuncture on learning and memory abilities in vascular dementia rats via the NCOA4/FTH1 signaling pathway-mediated ferritinophagy.
Wei SUN ; Yinghua CHEN ; Tong WU ; Hongxu ZHAO ; Haoyu WANG ; Ruiqi QIN ; Xiaoqing SU ; Junfeng LI ; Yuanyu SONG ; Yue MIAO ; Xinran LI ; Yusheng HAN
Chinese Acupuncture & Moxibustion 2025;45(9):1271-1280
OBJECTIVE:
To observe the effect of electroacupuncture at "Sishencong" (EX-HN1) and "Fengchi" (GB20) on hippocampal neuronal ferritinophagy mediated by the nuclear receptor coactivator 4 (NCOA4)/ferritin heavy chain 1 (FTH1) signaling pathway in vascular dementia (VD) rats, and to explore the potential mechanisms of electroacupuncture for VD.
METHODS:
A total of 60 male rats of SPF grade were randomly divided into a blank group (12 rats), a sham surgery group (12 rats) and a modeling group (36 rats). In the modeling group, the modified 4-vessel occlusion method was used to establish the VD model. The 24 successfully modeled rats were randomly divided into a model group and an electroacupuncture group, with 12 rats in each group. In the electroacupuncture group, electroacupuncture was applied at left and right "Sishencong" (EX-HN1), and bilateral "Fengchi" (GB20), with continuous wave, in frequency of 2 Hz and current intensity of 1 mA, 30 min a time, once daily for 21 consecutive days. The learning and memory abilities were assessed using the Morris water maze test before modeling, after modeling and after intervention, as well as the novel object recognition test after intervention. After intervention, the neuronal morphology in the hippocampus was observed by Nissl staining; the iron deposition was observed by Prussian blue staining; the reactive oxygen species (ROS) level was detected by dihydroethidium (DHE) fluorescence staining; the levels of iron, malondialdehyde (MDA) and superoxide dismutase (SOD) in the hippocampal tissue were measured by the colorimetric assay, TBA method, and WST-1 method, respectively; the positive expression of NCOA4, FTH1 and glutathione peroxidase 4 (GPX4) was detected by immunohistochemistry; the protein expression of NCOA4, FTH1, GPX4, and the ratio of microtubule-associated protein 1 light chain 3B (LC3B) Ⅱ/Ⅰ in the hippocampus were detected by Western blot.
RESULTS:
Compared with the sham surgery group, in the model group, the escape latency was prolonged, and the number of platform crossings reduced (P<0.01), the recognition index (RI) was decreased (P<0.01); the hippocampal neurons displayed a blurred laminar structure, disorganized cellular arrangement, and the number of Nissl bodies was decreased (P<0.01); the percentage of iron deposition area in the hippocampus was increased (P<0.01); in the hippocampus, the levels of ROS, iron, MDA, and the protein expression of NCOA4, as well as the LC3B Ⅱ/Ⅰ ratio were increased (P<0.01), the SOD level, and the protein expression of FTH1 and GPX4 were decreased (P<0.01). Compared with the model group, in the electroacupuncture group, the escape latency was shortened and the number of platform crossings was increased (P<0.01), the RI was increased (P<0.01); the hippocampal neurons exhibited more regular morphology, better-organized cellular structure, and the number of Nissl bodies was increased (P<0.05); the percentage of iron deposition area in the hippocampus reduced (P<0.01); in the hippocampus, the levels of ROS, iron, MDA, and the protein expression of NCOA4, as well as the LC3B Ⅱ/Ⅰ ratio were decreased (P<0.01, P<0.05), the SOD level, and the protein expression of FTH1 and GPX4 were increased (P<0.01).
CONCLUSION
Electroacupuncture at "Sishencong" (EX-HN1) and "Fengchi" (GB20) can improve learning and memory abilities in VD rats, and its mechanism may be associated with the regulation of the hippocampal NCOA4/FTH1 signaling pathway, inhibition of ferritinophagy, and alleviation of oxidative stress damage.
Animals
;
Electroacupuncture
;
Dementia, Vascular/genetics*
;
Male
;
Rats
;
Signal Transduction
;
Humans
;
Memory
;
Rats, Sprague-Dawley
;
Nuclear Receptor Coactivators/genetics*
;
Ferritins/genetics*
;
Learning
;
Hippocampus/metabolism*
;
Acupuncture Points
3.Study on the Lipid-Regulating Mechanism of Zexie Decoction on Hyperlipidemia Model Mice Based on the miR21/PI3K-Akt/SREBP Pathway
Aixia JU ; Shuang SUN ; Ruinan WANG ; Yubo HAN ; Yusheng ZHOU ; Jing CHEN ; Li LIU ; Qiuhong LI
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(9):917-931
OBJECTIVE To explore the lipid-regulating mechanism of the classic prescription Zexie Decoction on hyperlipidemia model mice.METHODS ELISA method was used to detect the four blood lipid indexes,liver function indicators and cholesterol acyltransferase levels in serum.HE and Oil Red O staining were used to determine the pathology of liver tissue.Network pharmacology was used to predict the lipid-lowering related targets of Zexie Decoction,and the GO function and KEGG pathway enrichment analyses of the intersection targets were realized.PCR chip technology was used to detect the target genes for network pharmacology screening,and qPCR and Western blot were used to detect gene and protein expression levels.RESULTS Zexie Decoction significantly regula-ted the four blood lipid indexes in hyperlipidemia model mice,improved the increase in liver damage indicators caused by high lipids,and had a reverse regulatory effect on the key enzymes HMGR and CYP7A1 of lipid metabolism and the lipid transporters ABCA1 and Apo-A1 in liver tissue.HE and Oil Red O staining showed that Zexie Decoction improved the pathological morphology of liver tissue,reduced lipid deposition in liver tissue,and significantly decreased the positive area ratio(P<0.01).The PCR chip obtained 44 re-verse-regulated genes,GO functional enrichment analysis obtained 266 entries,and KEGG pathway enrichment analysis screened 99 signaling pathways.The results of qPCR and Western blot showed that Zexie Decoction significantly downregulated the expression of PIK3CG,AKT1,and IL-6 genes(P<0.05,P<0.01),upregulated the expression of ABCG1 gene(P<0.05),downregulated PI3Kinase p110β,p-AKT(Ser473)and SREBP-1c protein expression levels(P<0.01),and reversely regulated miR21-5p(P<0.01).CONCLUSION Zexie Decoction has a significant regulatory effect on lipid metabolism in hyperlipidemia model mice and can improve liver damage caused by hyperlipidemia.Its lipid-regulating effect may be related to regulating cholesterol metabolism and transport in the body,and is closely linked to the miR21/PI3K-Akt/SREBP pathway.The lipid-regulating effect of the whole formula of Zexie Decoction is better than that of a single herb.
4.Investigation and analysis of the current status of transjugular intrahepatic portosystemic shunt treatment for portal hypertension in China
Haozhuo GUO ; Meng NIU ; Haibo SHAO ; Xinwei HAN ; Jianbo ZHAO ; Junhui SUN ; Zhuting FANG ; Bin XIONG ; Xiaoli ZHU ; Weixin REN ; Min YUAN ; Shiping YU ; Weifu LYU ; Xueqiang ZHANG ; Chunqing ZHANG ; Lei LI ; Xuefeng LUO ; Yusheng SONG ; Yilong MA ; Tong DANG ; Hua XIANG ; Yun JIN ; Hui XUE ; Guiyun JIN ; Xiao LI ; Jiarui LI ; Shi ZHOU ; Changlu YU ; Song HE ; Lei YU ; Hongmei ZU ; Jun MA ; Yanming LEI ; Ke XU ; Xiaolong QI
Chinese Journal of Radiology 2024;58(4):437-443
Objective:To investigate the current situation of the use of transjugular intrahepatic portosystemic shunt (TIPS) for portal hypertension, which should aid the development of TIPS in China.Methods:The China Portal Hypertension Alliance (CHESS) initiated this study that comprehensively investigated the basic situation of TIPS for portal hypertension in China through network research. The survey included the following: the number of surgical cases, main indications, the development of Early-TIPS, TIPS for portal vein cavernous transformation, collateral circulation embolization, intraoperative portal pressure gradient measurement, commonly used stent types, conventional anticoagulation and time, postoperative follow-up, obstacles, and the application of domestic instruments.Results:According to the survey, a total of 13 527 TIPS operations were carried out in 545 hospitals participating in the survey in 2021, and 94.1% of the hospital had the habit of routine follow-up after TIPS. Most hospitals believed that the main indications of TIPS were the control of acute bleeding (42.6%) and the prevention of rebleeding (40.7%). 48.1% of the teams carried out early or priority TIPS, 53.0% of the teams carried out TIPS for the cavernous transformation of the portal vein, and 81.0% chose routine embolization of collateral circulation during operation. Most of them used coils and biological glue as embolic materials, and 78.5% of the team routinely performed intraoperative portal pressure gradient measurements. In selecting TIPS stents, 57.1% of the hospitals woulel choose Viator-specific stents, 57.2% woulel choose conventional anticoagulation after TIPS, and the duration of anticoagulation was between 3-6 months (55.4%). The limitation of TIPS surgery was mainly due to cost (72.3%) and insufficient understanding of doctors in related departments (77.4%). Most teams accepted the domestic instruments used in TIPS (92.7%).Conclusions:This survey shows that TIPS treatment is an essential part of treating portal hypertension in China. The total number of TIPS cases is far from that of patients with portal hypertension. In the future, it is still necessary to popularize TIPS technology and further standardize surgical indications, routine operations, and instrument application.
5.Visual Analysis of Acupuncture Treatment of Vitiligo Based on CiteSpace
Li TANG ; Yusheng HAN ; Xianwei HAN
Journal of Zhejiang Chinese Medical University 2024;48(7):886-894
[Objective]To explore the research status and trend of acupuncture treatment of vitiligo for nearly 20 years,and provide certain reference for future research in this field.[Methods]The literatures related to acupuncture treatment of vitiligo were retrieved in China National Knowledge Infrastructure(CNKI),VIP Database and Wanfang Database from January 1,2003 to October 20,2023.After screening,CiteSpace software was used to visually analyse the literature authors,institutions and keywords.[Results]A total of 351 documents were included,and the number of articles published showed an upward trend,especially an explosive growth in 2017.A total of 297 authors were involved,and the core authors with the largest number of publications were REN Yingying,CHENG Aihua and LIU Hongxia,with 5 publications each.A small-scale cooperative team was formed among the authors.Shandong University of Traditional Chinese Medicine published the most papers,with 19 articles.There were 19 keywords with frequency≥5,involving 11 cluster labels and 15 emergent words.[Conclusion]In the past 20 years,the popularity of acupuncture therapy for vitiligo has increased,mainly focusing on the clinical study of fire acupuncture,and there are few basic researches.Large-scale and high-quality multi-center randomized controlled trials and basic experiments are carried out,and strengthening the cooperation between various teams and institutions is conducive to the further development of this field.
6.Exploration of electroacupuncture at "Fengchi" (GB 20) and "Sishencong" (EX-HN 1) for attenuating learning and memory impairment in vascular dementia rats based on NMDAR/CREB/BDNF signaling pathway.
Yuanyu SONG ; Yinghua CHEN ; Wei SUN ; Changqing LI ; Junfeng LI ; Haoyu WANG ; Ruiqi QIN ; Xiaoqing SU ; Tong WU ; Hongxu ZHAO ; Yusheng HAN
Chinese Acupuncture & Moxibustion 2024;44(12):1409-1417
OBJECTIVE:
To explore the mechanism of electroacupuncture (EA) at "Fengchi" (GB 20) and "Sishencong" (EX-HN 1) on learning and memory impairment in vascular dementia (VD) rats by observing the influences on the N-methyl-D-aspartate receptor (NMDAR)/cyclic adenosine monophosphate response element-binding protein (CREB)/brain-derived neurotrophic factor (BDNF) signaling pathway and the excitotoxicity induced by hippocampal calcium overload.
METHODS:
Thirty-two male SD rats of SPF grade were selected and randomized into a normal group (6 rats), a sham-operation group (6 rats) and an operation group (20 rats). VD model was established with the modified Pulsinelli's four-vessel occlusion (4-VO) method. Twelve rats after successfully modeled were assigned randomly into a model group and an EA group, 6 rats in each one. In the EA group, EA was delivered at bilateral "Fengchi" (GB 20) and "Sishencong" (EX-HN 1), with the continuous wave, the frequency of 2 Hz and the electric current of 1 mA. Stimulation intensity was adjusted depending on the slightly trembling of rat head. EA was given once daily, 30 min each time; and EA intervention was delivered for 21 days continuously. Using Morris water maze test, the learning and memory function was assessed. The neuronal morphology in the hippocampal CA1 was observed with HE staining; the level of glutamate (GLU) in serum and hippocampal tissue, as well as the activity of calcium pump (Ca2+-ATP) in the hippocampus were detected using colorimetric method. The protein expression of NMDAR, calmodulin-dependent protein kinase Ⅱ (CaMKⅡ), phosphorylated calmodulin-dependent protein kinase Ⅱ (p-CaMKⅡ), phosphorylated cyclic phosphoradenosine effector element binding proteins (p-CREB), CREB, and BDNF in the hippocampal CA1 was detected using immunohistochemistry. The protein expression of NMDAR, CREB, p-CREB and BDNF in the hippocampal tissue was detected using Western blot method.
RESULTS:
Compared to the sham-operation group, in the model group, the escape latency was prolonged and the platform crossing times of rats were reduced (P<0.01), the hippocampal neuron structure was damaged to different degrees, the structure in hippocampal CA1 was loosened, the arrangement disorganized, with clear grid-like structure; the neuronal morphology was irregular, pyknosis and even dissolution occurred, glial cells increased, blood capillary was dilated and the inflammatory cells were infiltrated and scattered. The level of GLU in the serum and hippocampal tissue and the protein expression of hippocampal NMDAR were elevated (P<0.01), the activity of Ca2+-ATP and the protein expression of CaMKⅡ, p-CaMKⅡ, CREB, p-CREB and BDNF were reduced (P<0.01, P<0.05); and the ratio of p-CaMKⅡ/CaMKⅡ and that of p-CREB/CREB were dropped (P<0.05). In comparison with the model group, in the EA group, the escape latency was shortened and the platform crossing times of rats rose (P<0.01), the arrangement was improved in the hippocampal CA1, the neuronal morphology was intact, the nucleoli were clear relatively and the pyknosis or dissolution were attenuated, the numbers of glial cells reduced relatively, the dilation of blood capillary was alleviated. The level of GLU in the serum and hippocampal tissue and the protein expression of NMDAR were reduced in the hippocampal tissue (P<0.01), the activity of Ca2+-ATP and the protein expression of CaMKⅡ, p-CaMKⅡ, CREB, p-CREB and BDNF were elevated (P<0.05, P<0.01); and the ratio of p-CaMKⅡ/CaMKⅡ and that of p-CREB/CREB increased (P<0.05).
CONCLUSION
EA at "Fengchi" (GB 20) and "Sishencong" (EX-HN 1) can attenuate learning and memory impairment in VD rats, which may be obtained by reducing GLU level in hippocampal tissue, inhibiting hippocampal excitotoxicity, mediating protein expression related to the NMDAR/CREB/BDNF signaling pathway, and maintaining neuronal survival and growth.
Electroacupuncture
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Male
;
Animals
;
Rats, Sprague-Dawley
;
Learning
;
Memory
;
Signal Transduction
;
Cyclic AMP Response Element-Binding Protein/metabolism*
;
Memory Disorders/therapy*
;
Brain-Derived Neurotrophic Factor/metabolism*
;
Receptors, N-Methyl-D-Aspartate/metabolism*
;
Dementia, Vascular/therapy*
7.Regulatory effect and mechanism of total alkaloids of Rhizoma Corydalis on hippocampal silencing information regulator 1/tumor suppressor P53 protein signaling pathway in rats with chronic cerebral ischemia
Jun LI ; Yazhi QI ; Ya TANG ; Rui CAO ; Yanling ZHAI ; Yusheng HAN ; Qiang XU
Chinese Journal of Cerebrovascular Diseases 2023;20(12):837-845
Objective To investigate the effect of total alkaloids of Rhizoma Corydalis(TAC)on the expression of silencing information regulator 1(Sirt1)/tumor suppressor P53 protein signaling pathway-related proteins in the hippocampus of rats with chronic cerebral ischemia(CCH),and to explore its mechanism.Methods The rats were randomly divided into Sham operation group,model group,TAC high-dose group(14 mg/kg)and TAC low-dose group(7 mg/kg),with 6 rats in each group.A modified bilateral common carotid artery permanent occlusion method(BCCAO)was used to establish a rat model of CCH,and only bilateral common carotid arteries were separated in the Sham group.After the modeling was completed,each group was given the corresponding drug or isotonic saline by gavage,once a day,and the treatment lasted for 14 days.Hematoxycin-eosin staining was used to observe the pathological changes in the hippocampus of rats,in situ terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphate-biotin nick end labeling assay(TUNEL)was used to detect neuronal apoptosis,and Western blotting and immunohistochemistry were used to detect expression of Sirt1,P53,P53 positive apoptosis regulator(PUMA),B-cell lymphocytoma-2(Bcl-2)protein,Bcl-2-related X protein(BAX),respectively in the hippocampus of rats.Results(1)There were significant differences in the number of apoptotic cells and apoptosis rate among the four groups(F-values were 71.417 and 76.835,respectively,both P<0.01).There were statistically significant differences in the mean integral optical density values of Sirt1,P53,PUMA,BAX and Bcl-2 protein positive expression areas among the four groups(F-values were 1 178.390,42.465,867.413,110.656 and 131.801,all P<0.01).There were significant differences in the relative expression levels of Sirt1,P53,PUMA,BAX and Bcl-2 among the four groups(F-values were 9.497,11.863,58.552,186.855 and 12.466,all P<0.01).(2)Compared with the Sham operation group,the neuronal arrangement of brain tissue in the hippocampus of the model group was disordered,the nuclear consolidation increased,and the glial cells and inflammatory cells increased significantly,and the number and apoptosis rate of neurons in the hippocampus of the model group increased significantly(respectively[10.8±1.5]cells vs.[2.0±0.9]cells and[35.5±4.5]%vs.[6.2±2.6]%;both P<0.05),and the average integral optical density values of the positive expression areas of Sirt1 and Bcl-2 proteins decreased significantly(84.6±6.6 vs.244.6±4.9,138.5±6.7 vs.210.9±10.0;both P<0.05),the average integral optical density values of P53,PUMA and BAX proteins were significantly increased(156.8±11.6 vs.93.5±11.6,151.3±3.3 vs.38.0±4.0,87.0±5.0 vs.38.4±5.5;all P<0.05),the relative expression levels of Sirt1 and Bcl-2 proteins were significantly decreased(0.51±0.07 vs.0.74±0.07,0.36±0.03 vs.0.53±0.05;both P<0.05),and the relative expression levels of P53,PUMA and BAX proteins were significantly increased(0.37±0.06 vs.0.21±0.02,0.62±0.06 vs.0.23±0.02,1.08±0.06 vs.0.45±0.03;all P<0.05).(3)Compared with the model group,the hippocampal tissue structure of the high-dose and low-dose TAC groups was relatively compact and uniform,the neurons were neatly arranged,and the cell structure was relatively clear and complete,while the number of neuronal apoptotic cells and the apoptosis rate decreased significantly(respectively[3.8±0.7]cells vs.[6.2±1.2]cells,[12.4±2.8]%vs.[20.2±3.9]%;both P<0.05),and the average integrated optical density values of the positive expression areas of Sirt1 and Bcl-2 proteins(the high-dose and low-dose TAC groups:Sirt1 150.0±4.8,131.3±1.3,and Bcl-2 207.1±7.4,169.5±3.9,respectively)were significantly increased(both P<0.05),the average integral optical density values of P53,PUMA and BAX proteins were significantly decreased(the high-dose and low-dose TAC groups:P53 105.9±8.8,115.5±9.0,and PUMA56.8±5.1,74.4±3.9,and BAX40.5±5.6,48.4±5.0,respectively,all P<0.05),the relative expression levels of Bcl-2 protein(the high-dose and low-dose TAC groups:0.53±0.05,0.47±0.02,respectively)were significantly increased(P<0.05),the relative expression levels of P53(the high-dose and low-dose TAC groups:0.21±0.02,0.24±0.04,respectively),PUMA(the high-dose and low-dose TAC groups:0.36±0.02,0.28±0.04,respectively)and BAX proteins(the high-dose and low-dose TAC groups:0.52±0.02,0.54±0.03,respectively)were significantly decreased(all P<0.05),the relative expression level of Sirt1 protein in the TAC high-dose group was significantly decreased(0.71±0.05,P<0.05),and the relative expression level of Sirt1 protein in the TAC low-dose group was not statistically significant(0.52±0.08,P>0.05).Conclusion TAC can alleviate neuronal damage and reduce the apoptosis rate of neurons in the hippocampus of CCH rats,and the mechanism may be related to the activation of Sirt1/P53 pathway,inhibition of P53 protein activity,and thus the expression level of apoptosis-related proteins in the downstream of TAC.
8.Dynamic DNA 5-hydroxylmethylcytosine and RNA 5-methycytosine Reprogramming During Early Human Development.
Xiao HAN ; Jia GUO ; Mengke WANG ; Nan ZHANG ; Jie REN ; Ying YANG ; Xu CHI ; Yusheng CHEN ; Huan YAO ; Yong-Liang ZHAO ; Yun-Gui YANG ; Yingpu SUN ; Jiawei XU
Genomics, Proteomics & Bioinformatics 2023;21(4):805-822
After implantation, complex and highly specialized molecular events render functionally distinct organ formation, whereas how the epigenome shapes organ-specific development remains to be fully elucidated. Here, nano-hmC-Seal, RNA bisulfite sequencing (RNA-BisSeq), and RNA sequencing (RNA-Seq) were performed, and the first multilayer landscapes of DNA 5-hydroxymethylcytosine (5hmC) and RNA 5-methylcytosine (m5C) epigenomes were obtained in the heart, kidney, liver, and lung of the human foetuses at 13-28 weeks with 123 samples in total. We identified 70,091 and 503 organ- and stage-specific differentially hydroxymethylated regions (DhMRs) and m5C-modified mRNAs, respectively. The key transcription factors (TFs), T-box transcription factor 20 (TBX20), paired box 8 (PAX8), krueppel-like factor 1 (KLF1), transcription factor 21 (TCF21), and CCAAT enhancer binding protein beta (CEBPB), specifically contribute to the formation of distinct organs at different stages. Additionally, 5hmC-enriched Alu elements may participate in the regulation of expression of TF-targeted genes. Our integrated studies reveal a putative essential link between DNA modification and RNA methylation, and illustrate the epigenetic maps during human foetal organogenesis, which provide a foundation for for an in-depth understanding of the epigenetic mechanisms underlying early development and birth defects.
Humans
;
DNA Methylation
;
RNA
;
Epigenesis, Genetic
;
DNA/genetics*
;
Basic Helix-Loop-Helix Transcription Factors/genetics*
9.CXCR4/SDF-1 axis regulates the effect of human lung adenocarcinoma PC-9 cells on function of in vitro blood-brain barrier model formed by Bends cells
LI Hongru ; TU Xunwei ; CHEN Zhengwei ; CHEN Yusheng ; HAN Lili
Chinese Journal of Cancer Biotherapy 2020;27(5):528-533
[Abstract] Objective: To investigate the influences of human lung adenocarcinoma PC-9 cells on tight junction proteins of blood brain barrier (BBB) under CXCR4/SDF-1 axis by establishing a model of BBB in vitro. Methods: The immortalized mouse brain microvascular endothelial Bends cells were used to establish a model of BBB in vitro by monolayer culture; Subsequently, transendothelial electric resistance (TEER) and fluorescein sodium permeability experiment were used to detect the function of in vitro BBB model and observe the effect of PC-9 cells on the function of BBB model, respectively. Western blotting was used to detect the effect of PC-9 cells on function of BBB model and expressions of endothelial tight junction proteins under the treatment of single or combined AMD3100 and SDF-1 (1 μg/ml AMD3100,100 ng/ml SDF-1, AMD3100+SDF-1). Transwell assay was used to detect the influence of CXCR4/SDF-1 axis on the ability of PC-9 cells transmigrating the cell layer of BBB model. Results: Monolayer culture of Bends cells can form tightly connected BBB withhighTEER,which reached (182.13±5.19) Ω.cm2 at the 96 h; in the meanwhile, fluorescein sodium permeability experiment showed that BBB had significantly lower permeability than that of control group ([40.31±2.43]% vs [150.10±3.17]%, P<0.05). The TEER of BBB decreased to (46.7±4.35) Ω·cm2 after coculture with PC-9 cells for 24 h, and at the same time the sodium fluorescein permeability of BBB significantly increased than that of pre-treatment ([136.32±4.93]% vs [50.24±6.21]%, P<0.05). PC-9 cells up-regulated the expressions of tight junction proteins of Bends cells under the treatment of AMD3100 (P<0.05). The number of PC-9 cells transmigrating the BBB inAMD3100 treatment group was significantly lower than that of CON group (43±2 vs 81±2, P<0.05). Conclusion: AMD3100 can reduce the ability of PC-9 cells destroying the tight junction of the BBB model established in vitro by Bends cells.
10.Effects of Dihydroquercetin on Hemorheology and Other Relevant Indexes in Local Cerebral Ischemic Injury Model Rats
Yuanyuan GU ; Bo JIANG ; Ming TIAN ; Yusheng HAN ; Xu LIU ; Yanqi SHANG ; Hui LIANG ; Xiaohong DONG ; Qiaomei DAI ; Zhongguang ZHOU
China Pharmacy 2019;30(6):765-769
OBJECTIVE: To observe the effects of dihydroquercetin (DHQ) on hemorheology and other relevant related indexes in local cerebral ischemic injury model rats. METHODS: SD rats were randomly divided into sham operation group, model group, nimodipine group (positive control, 20 mg/kg) and DHQ low-dose, medium-dose and high-dose groups (15, 30, 60 mg/kg), with 10 rats in each group. Administration groups were given relevant medicine intragastrically, sham operation group and model group were given constant volume of 0.4% Sodium carboxymethyl cellulose solution, once a day, for consecutive 14 d. After last administration, local cerebral ischemic injury model was induced by bilateral common carotid artery ligation in other groups except for sham operation group. After 24 h of cerebral ischemia, histopathological changes of brain tissue in rats of each group were observed; the levels of hemorheology indexes [whole blood viscosity (low, medium and high shear), whole blood reduced viscosity (low, medium and high shear), plasma viscosity], erythrocyte parameters (hematocrit, EAI, DI, IR), coagulation function indexes (APTT, PT, TT, FIB) were detected. RESULTS: Compared with sham operation group, the cells in the brain tissue of model group were loose, the gap was obvious, and the neurons around the ischemic area were damaged obviously; the levels of whole blood viscosity, whole blood reduced viscosity, plasma viscosity, hematocrit, EAI, IR and FIB were increased significantly, while the levels of DI, APTT, PT and TT were decreased or shortened significantly (P<0.05 or P<0.01). Compared with model group, above symptoms of administration groups were improved to different extents, whole blood viscosity, plasma viscosity, EAI and IR of nimodipine group, whole blood viscosity and hematocrit of DHQ high-dose group, plasma viscosity and EAI of DHQ groups, and IR of DHQ medium-dose and high-dose groups were decreased significantly; DI, APTT, PT and TT of nimodipine group, DI, APTT and TT of DHQ groups and PT of DHQ high-dose group were increased or prolonged significantly (P<0.05 or P<0.01). There was no statistical significance in other indexes among those groups (P>0.05). CONCLUSIONS: DHQ can protect against local cerebral ischemic injury model rats, the mechanism of which may be associated with improving hemorheology indexes and coagulation function disorder.

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