Variation rules of main secondary metabolites in Hedysari Radix before and after rubbing strip / 中成药

AIM To investigate the variation rules of main secondary metabolites in Hedysari Radix before and after rubbing strip.METHODS UPLC-MS/MS was adopted in the content determination of formononetin,ononin,calycosin,calycosin-7-glucoside,medicarpin,genistein,luteolin,liquiritigenin,isoliquiritigenin,vanillic acid,ferulic acid,γ-aminobutyric acid,adenosine and betaine,after which cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis were used for chemical pattern recognition to explore differential components.RESULTS After rubbing strip,formononetin,calycosin,liquiritigenin and γ-aminobutynic acid demonstrated increased contents,along with decreased contents of ononin,calycosin-7-glucoside and vanillic acid.The samples with and without rubbing strip were clustered into two types,calycosin-7-glucoside,formononetin,γ-aminobutynic acid,vanillic acid,calycosin-7-glucoside and formononetin were differential components.CONCLUSION This experiment clarifies the differences of chemical constituents in Hedysari Radix before and after rubbing strip,which can provide a reference for the research on rubbing strip mechanism of other medicinal materials.

Functional mechanisms of WRKY transcription factors in regulating plant response to abiotic stresses / 生物工程学报

WRKYs is a unique family of transcription factors (TFs) in plants, and belongs to the typical multifunctional regulator. It is involved in the regulation of multiple signaling pathways. This type of transcription factor is characterized to contain about 60 highly conservative amino acids as the WRKY domain, and usually also has the Cys2His2 or Cys2His-Cys zinc finger structure. WRKYs can directly bind to the W-box sequence ((T)(T) TGAC (C/T)) in the promoter region of the downstream target gene, and activate or inhibit the transcription of the target genes by interacting with the target protein. They may up-regulate the expression of stress-related genes through integrating signal pathways mediated by abscisic acid (ABA) and reactive oxygen species (ROS), thus playing a vital role in regulating plant response to abiotic stresses. This review summarizes the advances in research on the structure and classification, regulatory approach of WRKYs, and the molecular mechanisms of WRKYs involved in response to drought and salt stresses, and prospects future research directions, with the aim to provide a theoretical support for the genetic improvement of crop in response to abiotic stresses.

High-throughput screening of SARS-CoV-2 main and papain-like protease inhibitors

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (Mpro) and papain like protease (PLpro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851Mpro inhibitors and 205 PLpro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both Mpro and PLpro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of Mpro inhibitors and over 20% of PLpro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 Mpro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.

The relationship between the characteristics of tears and the progression of Stanford type B aortic dissection after endovascular treatment / 中华外科杂志

Endovascular treatment of Stanford type B aortic dissection (type B dissection) has been widely used. There will be complications such as aortic dilatation, which will lead to poor prognosis of some patients. With more in-depth researches, it was found that there was a possible correlation between the prognosis of type B dissection and tears, such as the increasing of aortic diameter would be faster with longer tears, and the location of the tear will affect the thrombosis of the false lumen. Studies on hemodynamics have also found that different characteristics of tears of aortic dissection can cause changes in the pressure, blood flow rate and blood capacity in the true and false lumens recently. The hemodynamic changes can be used to predict the prognosis of type B dissection. The main characteristics of tears included the size, position, number of tears, residual tears and stent graft induced new entry. Describing the effect of tear characteristics on the development of type B dissection, can provide the basis for the clinical treatment and further research of type B dissection.

Antiarrhythmic active components in traditional Chinese medicine acting on potassium channels / 中国中药杂志

Arrhythmia is an external manifestation of cardiac electrophysiological disorder. It exists in healthy people and patients with various heart diseases, which is often associated with other cardiovascular diseases. The contraction and diastole of myocardium are inseparable from the movement of ions. There are many ion channels in the membrane and organelle membrane of myocardium. The dynamic balance of myocardial ions is vital in maintaining myocardial electrical homeostasis. Potassium ion channels that have a complex variety and a wide distribution are involved in the whole process of resting potential and action potential of cardiomyocytes. Potassium ion channels play a vital role in maintaining normal electrophysiological activity of myocardium and is one of the pathogenesis of arrhythmia. Traditional Chinese medicine(TCM)has unique advantages in treating arrhythmia for its complex active components and diverse targets. A large number of TCM preparations have definite effect on treating arrhythmia-related diseases, whose antiarrhythmic mechanism may be related to the effect on potassium channel. This article mainly reviewed the relevant studies on the active components in TCM acting on different potassium channels to provide references for clinical drug use and development.

Chinese Guideline on the Management of Polypoidal Choroidal Vasculopathy (2022) / 中国医学科学杂志(英文版)

Background In mainland China, patients with neovascular age-related macular degeneration (nAMD) have approximately an 40% prevalence of polypoidal choroidal vasculopathy (PCV). This disease leads to recurrent retinal pigment epithelium detachment (PED), extensive subretinal or vitreous hemorrhages, and severe vision loss. China has introduced various treatment modalities in the past years and gained comprehensive experience in treating PCV.Methods A total of 14 retinal specialists nationwide with expertise in PCV were empaneled to prioritize six questions and address their corresponding outcomes, regarding opinions on inactive PCV, choices of anti-vascular endothelial growth factor (anti-VEGF) monotherapy, photodynamic therapy (PDT) monotherapy or combined therapy, patients with persistent subretinal fluid (SRF) or intraretinal fluid (IRF) after loading dose anti-VEGF, and patients with massive subretinal hemorrhage. An evidence synthesis team conducted systematic reviews, which informed the recommendations that address these questions. This guideline used the GRADE (Grading of Recommendations, Assessment, Development, and Evaluation) approach to assess the certainty of evidence and grade the strengths of recommendations. Results The panel proposed the following six conditional recommendations regarding treatment choices. (1) For patients with inactive PCV, we suggest observation over treatment. (2) For treatment-na?ve PCV patients, we suggest either anti-VEGF monotherapy or combined anti-VEGF and PDT rather than PDT monotherapy. (3) For patients with PCV who plan to initiate combined anti-VEGF and PDT treatment, we suggest later/rescue PDT over initiate PDT. (4) For PCV patients who plan to initiate anti-VEGF monotherapy, we suggest the treat and extend (T&E) regimen rather than the pro re nata (PRN) regimen following three monthly loading doses. (5) For patients with persistent SRF or IRF on optical coherence tomography (OCT) after three monthly anti-VEGF treatments, we suggest proceeding with anti-VEGF treatment rather than observation. (6) For PCV patients with massive subretinal hemorrhage (equal to or more than four optic disc areas) involving the central macula, we suggest surgery (vitrectomy in combination with tissue-plasminogen activator (tPA) intraocular injection and gas tamponade) rather than anti-VEGF monotherapy. Conclusions Six evidence-based recommendations support optimal care for PCV patients' management.

IgG78-DM1 inhibits pulmonary fibrosis by targeting and killing CD248-positive myofibroblasts in mice / 细胞与分子免疫学杂志

Objective To investigate the therapeutic effect of targeting and killing CD248-positive myofibroblasts on bleomycin-induced pulmonary fibrosis in mice. Methods IgG78-DM1, an antibody-maytansine 1 (DM1) conjugate targeting CD248, was prepared. The drug conjugation efficiency was measured and calculated by UV spectrophotometer, and the identification of IgG78-DM1 was performed through SDS-PAGE and Western blot analysis. In vitro, the binding activity of IgG78-DM1 on CD248-positive myofibroblasts was detected by flow cytometry and the cytotoxicity of IgG78-DM1 to CD248-positive myofibroblasts was evaluated by CCK-8 assay. In vivo, C57BL/6 male mice were randomly divided into control group, idiopathic pulmonary fibrosis group, human IgG-DM1 (hIgG-DM1) control group, and IgG78-DM1 treatment group. Then, the mouse models with pulmonary fibrosis induced by bleomycin were constructed. Two weeks later, the animal models were intravenously injected with IgG78-DM1. After the treatment of two weeks, lung tissues were collected for Masson staining and Sirius Red staining to evaluate the degree of pulmonary fibrosis. Real-time fluorescence quantitative PCR was used to measure the expression levels of CD248, as well as markers of fibroblastic activation including alpha-smooth muscle actin (α-SMA) and type I collagen alpha 1 (COL1A1). The safety of IgG78-DM1 was preliminarily assessed by conducting liver and kidney function tests. Results IgG78-DM1 was successfully prepared, and its drug conjugation ratio was 3.2. The antibody structure remained stable after conjugation, allowing effective binding and cytotoxicity against CD248-positive myofibroblasts. After treatment with IgG78-DM1, the degree of pulmonary fibrosis in mice significantly reduced, accompanied by the decrease of the expression of CD248, α-SMA, and COL1A1. The liver and kidney function of the mice remained at normal levels compared to the normal control group. Conclusion IgG78-DM1 effectively inhibits pulmonary fibrosis in mice by targeting and killing CD248-positive myofibroblasts. The safety of this strategy is preliminarily assessed.

Clinical effects of Qushi Zhitong Pills on patients with rheumatoid arthritis / 中成药

AIM To explore the clinical effects of Qushi Zhitong Pills on patients with rheumatoid arthritis.METHODS Eighty-six patients were randomly assigned into control group(43 cases)for 3-month intervention of conventional treatment,and observation group(43 cases)for 3-month intervention of both Qushi Zhitong Pills and conventional treatment.The changes in clinical efficacy,DAS28 score,laboratory-related indices(ESR,RF,CRP),peripheral blood Th17,Treg,Th17/Treg,bone destruction-related indices(OPG,BGP,PⅠNP,RANKL)and safety indices were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P<0.05).After the treatment,the two groups displayed decreased DAS28 score,laboratory-related indices,Th17,Th17/Treg,PⅠNP,RANKL(P<0.05),and increased Treg,OPG,BGP(P<0.05),especially for the observation group(P<0.05).No obvious adverse reactions were observable in the two groups.CONCLUSION For the patients with rheumatoid arthritis,Qushi Zhitong Pills can regulate Th17/Treg balance,improve bone destruction,and increase bone mineral density.

Design and application analysis of public health emergency training based on instructional system design model / 中华医学教育探索杂志

Objective:In order to improve the staff's ability to deal with public health emergencies, a third-class hospital in Beijing has made efforts to improve the emergency response ability of all staff members to achieve zero infection, and to carry out vocational training and assessment of all staff.Methods:The instructional system design (ISD) model system is used to design the training course. The online learning, electronic examination paper assessment and on-site training of people in the hospital were analyzed by Excel and SPSS 22.0.Results:After training, the average scores of trainees increased from 84 points to 100 points, and the average answering time was shortened from 308 s to 179 s. There were differences in the assessment scores before and after training for personnel with different professional titles, and there were differences in the assessment scores before and after training for personnel between different departments.Conclusion:In case of public health emergency, it is necessary to train and assess the whole staff. The application of ISD model is helpful to make training plan quickly. The application of online learning assessment is the first effective way of emergency training.

Psychosocial Work Conditions During the COVID-19 Pandemic and Their Influences on Mental Health Risk and Intention to Leave Among Public Health Workers: A Cross-sectional and Follow-up Study in Taiwan

Background@#To examine the influences of psychosocial work conditions on mental health risk and intention to leave the public sector among workers of public health agencies in Taiwan. @*Methods@#We surveyed 492 public health workers in March 2022 during the COVID-19 pandemic. Information on job demands, job control, workplace justice, experiences of workplace violence and its type and origin, and mental health status (assessed by the 5-item Brief Symptom Rating Scale, BSRS-5) was obtained. Of them, 192 participated in a follow-up survey conducted in May 2023 that assessed mental health status, employment changes, and intention to leave. @*Results@#In the initial survey, 32.93% of participants reported poor mental health status, defined by having a score of BSRS-5 ≧ 10, and 48.17% experienced some form of workplace violence over the past year. Notably, high psychosocial job demands (OR = 3.64, 95% CI = 1.93–6.87), low workplace justice (OR = 2.58, 95% CI = 1.45–4.58), and workplace violence (OR = 2.38, 95% CI = 1.51–3.77) were significantly associated with increased risk of mental disorders. Among those who participated in the follow-up survey, 22.40% had persistent poor mental health, and 30.73% considered leaving or have left the public sector. Longitudinal analyses indicated that job demands predicted persistent mental disorders and intention to leave the public sector, and the experience of workplace violence added additional mental health risks. @*Conclusion@#The public health workforce is crucial for effective and resilient public health systems. Our findings that public health workers were at high mental health risk and had a high intention to leave the job warrant attention and policy interventions.

Effect of boschniakia rossica polysaccharides on hypoxia/reoxygenation-induced cardiomyocyte damage by down-regulating miR-302a-3p expression / 中国药理学通报

Aim To explore the effect of boschniakia rossica polysaccharides ( BRPS ) on cardiomyocyte damage induced by hypoxia/reoxygenation (H/R) and its possible mechanism. Methods H/R was used to induce rat cardiomyocyte H9c2 to establish a cell inju¬ry model, and different doses of BRPS were used to treat H9c2 cells. ELISA method was used to detect the level of MDA and the activity of SOD and GSH-Px. Flow cytometry was used to detect the rate of apopto-sis. qRT-PCR was used to detect the expression of miR-302a-3p. anti-miR-NC and anti-miR-302a-3p were respectively transfected into H9c2 cells and then subjected to H/R treatment. miR-NC and miR-302a-3p mimics were respectively transfected into H9c2 cells and treated with 100 mg • L

m / 中国生物化学与分子生物学报

N6, 2′ -O-dimethyladenosine (m

Mechanism of Xianglian Pills in improving dyslipidemia in obese mice induced by high-fat diet based on network pharmacology and intestinal flora / 中国中药杂志

The present study aimed to investigate the effect of Xianglian Pills(XLP) on lipid metabolism in obese mice and explore the underlying mechanism based on network pharmacology and intestinal flora. Firstly, network pharmacology was used to predict the possible effect of XLP on obesity. Secondly, an obese mouse model induced by a high-fat diet was established to observe changes in mouse body weight, adiposity index, liver and adipose tissue pathology. Lipid profiles, liver and kidney function markers, insulin content, and the expression of recombinant uncoupling protein 1(UCP-1) and PR structural domain protein 16(PRDM16) were measured. The 16S rRNA gene sequencing technology was used to analyze the changes in the intestinal flora. Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis showed that XLP mainly played a role in improving obesity by regulating lipolysis, type 2 diabetes mellitus, and insulin resistance. The results of animal experiments showed that XLP significantly reduced body weight, adiposity, blood lipid levels, and serum insulin levels in obese mice, while enhancing the expression of UCP-1 and PRDM16 in adipose tissue without causing damage to the liver or kidneys. The 16S rRNA gene sequencing results showed that XLP decreased the Firmicutes/Bacteroidetes(F/B) ratio at the phylum level, increased the relative abundance of Akkermansia and Bacteroides at the family and genus levels, and reduced the abundance of Allobaculum. Therefore, XLP can effectively improve lipid metabolism disorders in high-fat diet-induced obese mice, and the mechanism is related to the improvement of brown adipose function, the browning of white fat, the accelerated lipid metabolism, and the improvement of intestinal flora. However, its effect on promoting the conversion of white adipose to brown adipose still needs to be further studied.

YULINK regulates vascular formation in zebrafish and HUVECs

BACKGROUND: The distinct arterial and venous cell fates are dictated by a combination of various genetic factors which form diverse types of blood vessels such as arteries, veins, and capillaries. We report here that YULINK protein is involved in vasculogenesis, especially venous formation. METHODS: In this manuscript, we employed gene knockdown, yeast two-hybrid, FLIM-FRET, immunoprecipitation, and various imaging technologies to investigate the role of YULINK gene in zebrafish and human umbilical vein endothelial cells (HUVECs). RESULTS: Knockdown of YULINK during the arterial-venous developmental stage of zebrafish embryos led to the defective venous formation and abnormal vascular plexus formation. Knockdown of YULINK in HUVECs impaired their ability to undergo cell migration and differentiation into a capillary-like tube formation. In addition, the phosphorylated EPHB4 was decreased in YULINK knockdown HUVECs. Yeast two-hybrid, FLIM-FRET, immunoprecipitation, as well as imaging technologies showed that YULINK colocalized with endosome related proteins (EPS15, RAB33B or TICAM2) and markers (Clathrin and RHOB). VEGF-induced VEGFR2 internalization was also compromised in YULINK knockdown HUVECs, demonstrating to the involvement of YULINK. CONCLUSION: This study suggests that YULINK regulates vasculogenesis, possibly through endocytosis in zebrafish and HUVECs. Key points Knockdown of YULINK with morpholino in embryos of double transgenic zebrafish exhibited abnormal venous formation. Tube formation and phosphorylated EPHB4 were decreased in YULINK knockdown HUVECs. FLIM-FRET, immunoprecipitation, as well as other imaging technologies showed that YULINK colocalized with endosome related proteins (EPS15, RAB33B and TICAM2) and endosome markers (Clathrin and RHOB). Knockdown of YULINK decreased the internalization of VEGF and VEGFR2 in HUVECs.

Modulating effects of RAMPs on signaling profiles of the glucagon receptor family

Receptor activity-modulating proteins (RAMPs) are accessory molecules that form complexes with specific G protein-coupled receptors (GPCRs) and modulate their functions. It is established that RAMP interacts with the glucagon receptor family of GPCRs but the underlying mechanism is poorly understood. In this study, we used a bioluminescence resonance energy transfer (BRET) approach to comprehensively investigate such interactions. In conjunction with cAMP accumulation, Gα q activation and β-arrestin1/2 recruitment assays, we not only verified the GPCR-RAMP pairs previously reported, but also identified new patterns of GPCR-RAMP interaction. While RAMP1 was able to modify the three signaling events elicited by both glucagon receptor (GCGR) and glucagon-like peptide-1 receptor (GLP-1R), and RAMP2 mainly affected β-arrestin1/2 recruitment by GCGR, GLP-1R and glucagon-like peptide-2 receptor, RAMP3 showed a widespread negative impact on all the family members except for growth hormone-releasing hormone receptor covering the three pathways. Our results suggest that RAMP modulates both G protein dependent and independent signal transduction among the glucagon receptor family members in a receptor-specific manner. Mapping such interactions provides new insights into the role of RAMP in ligand recognition and receptor activation.

Berberine alleviates programmed necrosis of metabolic-associated fatty liver disease via activating Nrf2 pathway in mice / 中华肝脏病杂志

Objective: To investigate the effect of berberine on programmed necrosis of hepatocytes induced by metabolic-associated fatty liver disease (MAFLD) in mice and its related molecular mechanism. Methods: Twenty male C57BL/6N mice were randomly divided into four groups (n=5 in each group): control group (S), fatty liver group (H), berberine group(B), nuclear factor erythroid 2-related factor 2 inhibitor group (Nrf2), and all-trans-retinoic acid (ATRA) group (A). Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), triglycerides (TG), total cholesterol (TC), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) concentrations were detected at the end of week 12 to calculate fatty liver index (liver mass/body mass ratio). Liver tissue was stained with HE, Masson and Oil Red O, and SAF score was used to evaluate the degree of liver injury. The expression levels of hepatic programmed necrosis-related proteins, namely receptor-interacting protein kinase 3 (RIPK3), phosphorylated mixed series protease-like domain (p-MLKL) and Nrf2 were detected by Western blot method. One-way ANOVA was used for intragroup comparisons and LSD-t tests were used for intergroup comparisons. Results: Compared with S group, H group serum ALT, AST, LDH, TG, TC, TNF-α, IL-1β levels and fatty liver index were significantly increased. The liver tissue was filled with vacuolar-like changes and inflammatory cell infiltration. Numerous red lipid droplets were observed with oil red O staining. Collagen fiber hyperplasia was evident with Masson staining. SAF scores (6.60 ± 0.55 and 0.80 ± 0.45) were significantly increased. The expressions of RIPK3 and p-MLKL were up-regulated. Nrf2 level was relatively increased, and the differences were statistically significant (P < 0.05). Compared with H group, berberine intervention group liver biochemical indexes, lipid levels, pro-inflammatory mediator expression, fatty liver index, and SAF score were significantly reduced, and the expression of RIPK3 and p-MLKL were down-regulated, while Nrf2 levels were further increased, and the differences were statistically significant (P<0.05). Compared with B group, treatment with Nrf2 inhibitor had antagonized the protective effect of berberine on fatty liver. Serum ALT, AST, LDH, TG, TC and TNF-α, IL-1β levels, fatty liver index, and SAF scores were significantly increased and the expressions of RIPK3 and p-MLKL were relatively increased, and the differences were statistically significant (P < 0.05). Conclusion: Berberine can significantly improve the metabolic-associated fatty liver disease injury in mice, and its mechanism is related to activation of Nrf2 and inhibition of programmed necrosis of hepatocytes.

The role of SnRK2 in the response to stress, the growth and development of plants / 生物工程学报

Sucrose non-fermenting-1-related protein kinase 2 (SnRK2) is a specific Ser/Thr protein kinase in plants. SnRK2 can regulate the expression of downstream genes or transcription factors through phosphorylation of substrates to achieve stress resistance regulation in different tissue parts, and make plants adapt to adverse environment. SnRK2 has a small number of members and a molecular weight of about 40 kDa, and contains a conserved N-terminal kinase domain and a divergent C-terminal regulatory domain, which plays an important role in the expression of enzyme. This review summarized the recent research progresses on the discovery, structure, and classification of SnRK2, and its function in response to various stresses and in regulating growth and development, followed by prospecting the future research direction of SnRK2. This review may provide a reference for genetic improvement of crop stress resistance.

Effect of propofol on proliferation of neural stem cells in mice and the role of Sp1-EGFR-Akt signaling pathway / 中华麻醉学杂志

Objective:To evaluate the effect of propofol on proliferation of neural stem cells (NSCs) in mice and the role of specificity protein-1 (Sp-1)-epidermal growth factor receptor (EGFR)-protein kinase B (Akt) signaling pathway.Methods:Primary NSCs harvested from both the cortices and hippocampus of C57BL/6 mouse embryos were identified by immunofluorescent staining of Nestin.NSCs at passages 3-6 were divided into 3 groups ( n=21 each) using a random number table method: normal saline control group (C group), propofol group (P group) and propofol plus Sp1 inhibitor plicamycin group (PP group). Propofol at a final concentration of 10 μmol/L was added in group P. Propofol at a final concentration of 10 μmol/L and plicamycin at a final concentration of 100 nmol/L were added in group PP.The equal volume of normal saline was added in group C. The medium was replaced after 6 h of incubation and the cells were continuously incubated.The proliferation of NSCs was assessed by direct cell counting at 24, 36, 48, 60 and 72 h after the end of treatment with drugs.At 6 h after the end of treatment with drugs, the expression of Sp1 and EGFR mRNA was detected by real-time fluorescent quantitative polymerase chain reaction, and the expression of Sp1, Akt and phosphorylated Akt (p-Akt) by Western blot. Results:Compared with group C, the count of NSCs was significantly increased at 48, 60 and 72 h after treatment with drugs, and the expression of EGFR mRNA, Sp1 protein and mRNA and p-Akt was up-regulated in group P ( P<0.05 or 0.01), and no significant change was found in each parameter in group PP ( P>0.05). Compared with group P, the count of NSCs was significantly decreased at 48 and 60 h after treatment with drugs, and the expression of EGFR protein and mRNA and p-Akt was down-regulated in group PP ( P<0.05 or 0.01). Conclusions:Propofol can promote the proliferation of NSCs, and the mechanism may be related to activation of Sp1-EGFR-Akt signaling pathway in mice.

The impact of excessive gestational weight gain on fetal hepatic lipid metabolism and the regulatory mechanism / 中华实用儿科临床杂志

Objective:A rat model of excessive gestational weight gain (EGWG) was constructed to investigate the impact of EGWG on fetal hepatic lipid metabolism and the relevant regulatory mechanism.Methods:Healthy Sprague-Dawley rats were caged together and tested for pregnancy.Rats with the sperm observed under microscope were considered pregnant for 0.5 days.Pregnant rats were divided into the normal diet (ND) group and high-fat diet (HFD) group by the random number table method, with 8 rats in each group.The body weight during pregnancy of the pregnant rats was recorded.Cesarean section was performed at day 21.5 of gestation and the birth weight of the fetal rats was recorded.Hepatic lipid deposition of the pregnant and fetal rats was examined by hematoxylin-eosin (HE) staining and oil red O staining.Triglyceride (TG) and cholesterol (TC) levels in livers and serum of the pregnant and fetal rats were detected by glycerol phosphate oxidase-peroxidase(GPO-PAP) method.The mRNA and protein expression levels of key genes FASN and SREBP1c in hepatic lipid metabolism of fetal rats were measured by real-time polyme-rase chain reaction (RT-PCR) and Western blot.Differences between the two groups were compared by independent sample t test. Results:There was no difference in pre-pregnancy body weight between the HFD group and the ND group, but the differences in the weight and the weight gain during pregnancy gradually enlarged between the two groups.At day 21.5 of gestation, the weight of the pregnant rats[(467.75±22.05) g vs.(430.88±18.80) g, t=-3.600, P=0.003], the weight gain of the pregnant rats during pregnancy[(181.50±9.68) g vs.(148.50±10.86) g, t=-6.415, P<0.001] and the birth weight of the fetal rats[(5.51±0.17) g vs.(4.85±0.35) g, t=-4.779, P<0.001] of the HFD group were significantly higher than those of the ND group.Both HE staining and oil red O staining presented increased hepatic lipid deposition in the pregnant and fetal rats of the HFD group.The hepatic and serum TG and TC levels of the pregnant and fetal rats of the HFD group were significantly higher than those of the ND group (all P<0.05). RT-PCR and Western blot showed that the mRNA and protein levels of key genes FASN and SREBP1c in hepatic lipid metabolism of fetal rats of the HFD group were significantly higher than those of the ND group (all P<0.05). Conclusions:An EGWG model can be successfully constructed by a 21-day HFD during pregnancy.EGWG can lead to hepatic lipid deposition in the fetal rats.The mechanism may be related to the expression changes of key genes FASN and SREBP1c in hepatic lipid metabolism of fetal rats.

Effects and mechanisms of different types of maternal obesity on glucose and lipid metabolism in neonatal offspring rats / 中国新生儿科杂志

Objective:To study the different effects of pre-pregnancy obesity (PO), excessive gestational weight gain (EGWG), pre-pregnancy obesity combined with excessive gestational weight gain (PO+EGWG) of maternal rats on glucose and lipid metabolism in neonatal offspring, and to explore the possible mechanisms.Methods:Animal models of PO, EGWG and PO+EGWG were established by feeding SD rats with high-fat diets at different periods. Thirty-six SD rats were randomly divided into four groups, with nine rats in each group. The control group had a normal diet before and during pregnancy. The PO group had a high-fat diet before pregnancy and a normal diet during pregnancy. The EGWG group had a normal diet before pregnancy and a high-fat diet during pregnancy. And the PO+EGWG group had a high-fat diet before and during pregnancy. The body weight of maternal rats before and during pregnancy and the birth weight of neonatal rats were recorded. Nine male neonatal rats in each group were selected, fasting blood glucose levels were detected by glucometer, fasting insulin levels were detected by enzyme-linked immunosorbent assay kit, hepatic triglyceride and cholesterol levels were detected by glycerol phosphate oxidase-peroxidase method, hepatic lipid deposition were observed by hematoxylin-eosin staining and oil red O staining. The mRNA levels of hepatic key genes in glucose metabolism pathway IR, IRS, AKT and lipid metabolism FASN, SREBP1c, PPARα were detected by reverse transcription-polymerase chain reaction analyses.Results:The pre-pregnancy weight of maternal rats in high-fat diet group before pregnancy (PO group and PO+EGWG group) was significantly higher than those in normal diet group (control group and EGWG group). The percentage of weight gain of maternal rats in high-fat diet group during pregnancy (EGWG group and PO+EGWG group) was significantly higher than those in normal diet group (control group and PO group) ( P<0.05). The birth weight of neonatal rats in PO group, EGWG group and PO+EGWG group were significantly higher than that in control group ( P<0.05), and the birth weight of neonatal rats in PO+EGWG group was the largest. The fasting glucose, insulin level and insulin resistance index of newborn rats in PO, EGWG and PO+EGWG groups were higher than those in the control group, and the mRNA levels of IR, IRS and AKT were lower than those in the control group, but the differences were not statistically significant ( P>0.05). The hepatic triglyceride and cholesterol contents and mRNA levels of FASN and SREBP1c were higher in the EGWG and PO+EGWG groups than those in the control group, and the mRNA level of PPARα was higher in the PO+EGWG group than in the control and PO groups, with statistically significant differences ( P<0.05). Conclusions:Animal models of PO, EGWG and PO+EGWG were successfully constructed by feeding SD rats with high-fat diets before pregnancy, during pregnancy, before and during pregnancy. PO+EGWG had the most significant effects on the birth weight and glucose and lipid metabolism in neonatal offspring. Compared with EGWG, PO had a relatively significant effect on glucose metabolism in neonatal offspring. And compared with PO, EGWG had a relatively significant effect on lipid metabolism in neonatal offspring. The effects of maternal obesity on glucose and lipid metabolism in neonatal offspring were considered to be related to the expression changes of genes in glucose and lipid metabolism.