1.Effect of Icariin on Myocardial Remodeling in Rats Based on Vitamin D Regulation of Dendritic Cell Phenotype
Qian LI ; Yujia CHEN ; Yan ZHOU ; Wen LI ; Liancheng GUAN ; Huanzhen WANG ; Yunzhi CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(5):76-85
ObjectiveTo investigate the effect of icariin (ICA) on the phenotype of dendritic cells (DCs) in heart tissue of the Dahl salt-sensitive myocardial remodeling model of rats and its regulation on the vitamin D system. MethodsMale Dahl salt-resistant rats were divided into a normal group, and male Dahl salt-sensitive rats were divided into a model group, low-, medium-, and high-dose ICA groups (30, 60, 120 mg·kg-1·d-1), and Vitamin D group (3×10-5 mg·kg-1·d-1). In addition to the normal group, the other groups were given an 8% high salt diet to establish a myocardial remodeling model and received intragastric administration after successful modelling once a day for six weeks. The dynamic changes in tail artery blood pressure were monitored, and detection of cardiac ultrasound function in rats was performed. Hematoxylin-eosin (HE) staining and Masson staining were used to observe the morphological changes in rat heart tissue. The phenotype of DCs and T helper cell 17 (Th17)/regulatory T cell (Treg) ratio were detected by flow cytometry. The mRNA and protein expression of vitamin D receptor (VDR), 1α-hydroxylase (CYP27B1), 24-hydroxylase (CYP24A1), forkhead frame protein 3 (FoxP3), solitaire receptor γt (RORγt), myocardial type Ⅰ collagen (ColⅠ), and type collagen (ColⅢ) in heart tissue was detected by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot. ResultsCompared with the normal group, the model group showed disordered arrangement and rupture of myocardial cells, nuclear condensation, significant edema of myocardial tissue, significant proliferation of collagen fibers in a network distribution, and a significant increase in tail artery blood pressure, left ventricular end diastolic diameter (LVEDD), and left ventricular end systolic diameter (LVESD) (P<0.05). The phenotype of cardiac DCs was CD40, CD80, and CD86, and the levels of major histocompatibility complex Ⅱ (MHC-Ⅱ), Th17 cells, and Th17/Treg were significantly increased (P<0.05). The mRNA and protein expression of CYP24A1 and RORγt in the heart, as well as the mRNA expression of ColⅠ and ColⅢ, were significantly increased (P<0.05). The left ventricular ejection fraction (LVEF), interventricular septal thickness (IVSD), and left ventricular posterior wall thickness (LVPWD) were significantly decreased (P<0.05). The phenotype of cardiac DCs such as CD11, CD11b, and Treg cells, were significantly reduced (P<0.05), while the mRNA and protein expression of cardiac VDR, CYP27B1, and FoxP3 were significantly decreased (P<0.05). Compared with the model group, the low-, medium-, and high-dose ICA groups and vitamin D group significantly reduced myocardial cell rupture and nuclear consolidation in rats. The high-dose ICA group and vitamin D group showed a small amount of myocardial cell rupture and nuclear consolidation, improving myocardial fiber arrangement to varying degrees and significantly reducing myocardial fiber rupture and proliferation. The tail artery blood pressure, LVEDD, and LVESD were significantly decreased in the low-, medium-, and high-dose ICA groups and vitamin D group (P<0.05), and the phenotype of cardiac DCs including CD40, CD80, CD86, MHC-Ⅱ, Th17 cells, and Th17/Treg were significantly decreased (P<0.05). The mRNA and protein expression of CYP24A1 and RORγt, and the mRNA expression of ColⅠ and ColⅢ in the heart were significantly decreased in the medium- and high-dose ICA groups and vitamin D group (P<0.05). The LVEF, IVSD, and LVPWD of myocardial remodeling model rats in the low-, medium-, and high-dose ICA groups and vitamin D group were significantly increased (P<0.05). The phenotypes of cardiac DCs including CD11, CD11b, and Treg cells were significantly increased in the medium- and high-dose ICA groups and the Vitamin D group (P<0.05). The mRNA and protein expressions of VDR, CYP27B1, and FoxP3 in the heart were significantly increased in the medium- and high-dose ICA groups and vitamin D group (P<0.05). ConclusionICA can regulate tail artery blood pressure, cardiac structural and functional damage, and myocardial tissue fibrosis and inhibit phenotype and functional maturation of DCs in heart tissue in the myocardial remodeling model of Dahl salt-sensitive rats. It can also affect the gene and protein expression of VDR, CYP24A1, and CYP27B1, achieving its intervention in Th17/Treg balance in the immune process of myocardial remodeling possibly by regulating vitamin D/VDR in heart tissue.
2.Relationship between peripheral blood CD4+/CD8+,D-dimer levels and the outcome of EBV-associated hemophagocytic syndrome
Yujia WANG ; Xin LI ; Deli SONG ; Chenlu WANG
Chinese Journal of Infection and Chemotherapy 2024;24(5):558-563
Objective To investigate the relationship between the levels of CD4+/CD8+and D-dimer(D-D)in peripheral blood and the outcome of Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis(EBV-HLH).Methods Ninety patients with EBV-HLH who were treated in Beijing Friendship Hospital from September 2021 to August 2023 were included as observation group,and additional 90 patients with infectious mononucleosis(IM)treated in the hospital contemporarily were selected as the control group.The levels of CD4+/CD8+and D-D in peripheral blood were compared between the two groups of patients to examine the correlation between the levels of CD4+/CD8+and D-D in peripheral blood and EBV-DNA load in patients with EBV-HLH,compare the outcome of patients within 3 months in terms of the levels of CD4+/CD8+and D-D in peripheral blood,evaluate the effects of CD4+/CD8+and D-D levels in peripheral blood on the risk of death from EBV-HLH,and analyze the interaction between the levels of CD4+/CD8+and D-D in peripheral blood.Results The patients in observation group showed significantly lower peripheral blood CD4+/CD8+ratio and significantly higher D-D than the patients in control group(P<0.05).The peripheral blood CD4+/CD8+ratio was negatively correlated with EBV-DNA load(P<0.05),and D-D was positively correlated with EBV-DNA load(P<0.05)in EBV-HLH patients.The patients with high peripheral blood CD4+/CD8+ratio were assocaited with lower 3-month mortality rate compared to the patients with low CD4+/CD8+ratio.The patients with high D-D level were associated with higher 3-month mortality rate compared to the patients with low D-D level(P<0.05).In EBV-HLH patients with low levels of peripheral blood CD4+/CD8+,the risk of death was 6.125 times that of the patients with high levels of CD4+/CD8+.High level D-D was associated with 14.348 times risk of death compared to the patients with low level D-D.CD4+/CD8+and D-D had synergistic effect on death of EBV-HLH patients.Conclusions Peripheral blood CD4+/CD8+levels decreased and D-D levels increased in EBV-HLH patients.Peripheral blood CD4+/CD8+levels and D-D levels and their changes may be useful for predicting the outcome of patients.
3.Mechanism of IL-6 improving pregnancy outcome in mice with recurrent spontaneous abortion
Rong WU ; Yuhong WENG ; Yujia LI ; Zirui WU ; Guanyou HUANG
Chinese Journal of Immunology 2024;40(8):1615-1619
Objective:To investigate whether IL-6 using in early pregnancy can improve the pregnancy outcome of recurrent spontaneous abortion(RSA)mice and its relevant mechanism,providing new ideas for RSA clinical treatment.Methods:CBA/J×DBA/2 RSA model mice were constructed,and randomly divided the pregnant mice into five groups:control group,0.1 ng/ml IL-6 group,1 ng/ml IL-6 group,10 ng/ml IL-6 group and 100 ng/ml IL-6 group.IL-6 was not injected in control group,while different concentra-tions of IL-6 were respectively injected into other groups on the 0.5 day of pregnancy.Pregnant rats were killed at 13.5 d and the embryo loss rate was calculated,the placental tissue was taked out,and expressions of IL-6 and indoleamine 2,3-dioxygenase(IDO)in tissues were detected by Western blot.Results:Embryo absorption rates of 0.1 ng/ml IL-6 group,1 ng/ml IL-6 group,10 ng/ml IL-6 group and 100 ng/ml IL-6 group were obviously lower than that in control group(P=0.002 4,P=0.007 0,P=0.027 0,P=0.031 0).IL-6 of exogenous injection was positive correlated with that expressed in mice placental tissue(r=0.791,P=0.000 052).IL-6 concentration of exogenous injection was between 0~1 ng/ml,which was positively correlated with IDO expression in placental tissue(r=0.868,P<0.000 1),IL-6 was positively correlated with IDO expressed in placental tissue(r=0.982,P<0.000 1).IL-6 concentration of exogenous injection was between 1~100 ng/ml,which was inversely correlated with IDO expression(r=-0.725,P=0.002),and IL-6 was inversely correlated with IDO expressed in placental tissue(r=-0.972,P<0.000 1).Conclusion:A single intraperitoneal injection of specific concentration of exogenous IL-6 to RSA mice can reduce embryo absorption rate of mice and modify their pregnancy outcome,which possible mecha-nism is the exogenous IL-6 induces expressions of IL-6 and IDO for a long term.Whether the IDO expression in placental tissue in-crease or not can be regarded as a mark for whether the specific concentration IL-6 can protect the pregnancy or not.
4.Effects of aerobic exercise on renal function abnormalities and mitochondrial oxidative stress of obese rats
Jianfang XU ; Yujia LIU ; Qing YANG ; Liang LI ; Bin ZHANG
Chinese Journal of Sports Medicine 2024;43(9):730-740
Objective To explore the effect of aerobic exercise on renal function abnormalities and mitochondrial oxidative stress of obese rats.Methods Ninety 5-week male Sprague-Dawley rats were randomly divided into a normal diet group(CON,n=10)and a high-fat diet group(n=80).The latter group was on high-fat diet for 8 weeks to induce obesity with renal function abnormality.After success-ful modelling,40 rats were chosen and randomly divided into a high-fat diet control group(HFD)and three groups of high-fat diet+aerobic exercise with different intensities(40%VO2max,60%VO2max,and 80%VO2max),each of 10.All exercise groups underwent daily 60-min aerobic exercise on treadmill,5 days per week for 4 weeks.After the intervention,their body weight,body length,perirenal and epi-didymal fat weights were measured,and morphometric indices including fat-body ratio and Lee's in-dex were calculated.Moreover,such biochemical indicators of renal function as serum creatinine(SCr),serum cystatin C and urinary microalbumin(mALB)were tested.Meanwhile,the pathological changes of the kidney were observed using hematoxylin eosin(HE)staining and Periodic acid-Schiff(PAS)staining,while the ultrastructural changes of the kidney and mitochondria were observed using the transmission electron microscopy.Moreover,the levels of superoxide dismutase(SOD),malondialde-hyde(MDA),and mitochondrial membrane potential in mitochondria were evaluated using ELISA kits.Results After 4 weeks of intervention,the average SCr,serum cystatin C,and mALB levels in the HFD group increased significantly compared with the CON group(P<0.05).Moreover,the average SCr levels of all exercise groups were significantly lower than the HFD group(P<0.05),with the average serum cystatin C and mALB levels of the 60%VO2max and 80%VO2max groups significantly lower than the HFD group(P<0.05).Meanwhile,the renal tubular epithelial cells in the HFD group showed mod-erate degeneration and increased glomerulosclerosis index(GSI)(P<0.05)and the degree of glomerular hypertrophy in each exercise group was significantly lower than the HFD group,with the renal tubules of the 60%VO2max group showing a clearer contour and decreased GSI(P<0.05).What's more,com-pared with the CON group,the number of swollen mitochondria in the HFD group increased,but the mitochondria damage of the 60%VO2max group relieved significantly compared with the HFD group.The renal mitochondrial MDA levels in the CON group increased significantly compared with the HFD group(P<0.05),and those of the exercise groups were significantly lower than the HFD group(P<0.05),with the most significant decrease in the 40%VO2max group(P<0.05).Moreover,the average renal mito-chondrial SOD and membrane potential levels in the HFD group decreased significantly compared with the CON group(P<0.05),while those in the 40%and the 60%VO2max groups were significantly higher than the HFD group(P<0.05).Conclusion Moderate to low intensity aerobic exercise can relieve renal dysfunction of obese rats by lowering the level of oxidative stress in renal mitochondria,mainly down-regulating such renal function biochemical indicators as SCr,serum cystatin C,and mALB,as well as alleviating the degree of renal tissue damage.
5.Expert consensus on postoperative care of patients with a left ventricular assistant device
Nursing Professional Committee of the National Cardiovascular Disease Expert Committee ; Yan MA ; Rong WU ; Chen ZHANG ; Qingyin LI ; Yujia HUANG ; Mingjing ZHAO ; Qiang FU ; Yonggang LI ; Jiani WANG
Chinese Journal of Nursing 2024;59(14):1687-1690
Objective To standardize nursing management on postoperative patients with a left ventricular assist-ant device(LVAD).Methods The first draft of the Consensus was formed on the basis of literature review.2 rounds of expert consultations and a round of online meeting discussion were held for adjustments and modifications the draft of the Consensus.Results The recovery rate of the inquiry questionnaire was 93.75%.The authority coefficients of the 2 rounds of inquiry experts were 0.927 and 0.920.The concentration degree of expert opinions for each indicator was greater than 3.5 score,and the coefficient of variation was less than 0.25.The Kendall harmony coefficients for 2 rounds of correspondence were 0.402 and 0.407(P<0.01).The final Consensus formed through expert consultations and meetings includes 7 themes:hemodynamic monitoring,LVAD function monitoring,coagulation function monitoring,percutaneous cable and wound care,exercise rehabilitation care,health education and guidance,and pre-discharge assessment.Conclusion The Consensus is scientific,rigorous,and authoritative.The Consensus covers all aspects of postoperative care for patients with LVAD,and it will benefit to clinical practice.
6.Analgesic management in hospice care
Tonghui FENG ; Xinyu CHEN ; Mengting GU ; Qiaozhen XIANG ; Yujia LI ; Yanfei XIA
Chinese Journal of Geriatrics 2024;43(7):797-801
Pain is a prevalent symptom in both cancer and non-cancer end-stage diseases, often being the most feared by patients and significantly impacting their quality of life.Hospice care aims to address physical, psychological, spiritual, and other needs of patients and their families during this stage, with a focus on alleviating pain and discomfort.Effective pain management is a crucial component of hospice care, particularly given the increasing prevalence of cancer and chronic diseases in China and the growing elderly population.To provide analgesic management for hospice patients, a thorough assessment of pain is essential to identify its type and characteristics.Treatment approaches may include etiological interventions, pharmacotherapy, interventional therapy, physiotherapy, psychotherapy, and comfort care, all aimed at achieving comprehensive pain management.The use of opioid should be carefully guided by scientific principles to minimize adverse effects and optimize pain relief, ultimately enhancing patients' end-of-life quality of life.
7.Development and validation of a prediction model for abnormal bone mass in end-stage renal disease patients
Jing LU ; Yujia WANG ; Yuxia ZHANG ; Zhiqing CHEN ; Yongqi LI ; Min WU ; Rining TANG
Chinese Journal of Nephrology 2024;40(5):345-357
Objective:To identify the risk factors, and develop and validate a risk prediction model for abnormal bone mass in end-stage renal disease (ESRD) patients.Methods:It was a retrospective cross-sectional study. The clinical and laboratory data of ESRD patients who were hospitalized in the Department of Nephrology, Zhongda Hospital Affiliated to Southeast University from January 2022 to May 2023 were collected retrospectively. The patients were randomly divided into training and validation cohorts at a ratio of 7∶3. They were further divided into normal and abnormal bone mass groups according to the T value measured by dual-energy X-ray absorptiometry (DXA). Then, backward stepwise regression and least absolute shrinkage and selection operator (LASSO) were respectively used to develop the risk prediction model for abnormal bone mass in ESRD patients. Akaike information criterion (AIC), bayesian information criterion (BIC), and accuracy were used to evaluate the performance of these two models, after which the preferable model was selected. Moreover, the receiver operating characteristic (ROC) curve, calibration curve, Hosmer-Lemeshow test, and decision curve analyses (DCA) were applied to evaluate the diagnostic performance of the preferable model. Finally, a dynamic nomogram for individual assessment was constructed based on the preferable model.Results:A total of 254 ESRD patients were enrolled, including 160 (63.0%) males, 161 (63.4%) hemodialysis patients, and 202 (79.5%) patients with abnormal bone mass. There was no significant difference in the prevalence of abnormal bone mass between training group ( n=178) and validation group ( n=76) (79.2% vs. 80.3%, χ2=0.036, P=0.849). The final variables and variable parameters included in the LASSO and stepwise regression models were the same, which were five variables: age, body mass index, hypertension, diabetes, and osteocalcin. Both models also had the same AIC, BIC, and accuracy in the training group, which were 113.45, 132.54, and 0.837, respectively. Therefore, the LASSO model and the stepwise regression model performed consistently in this study and could be considered as the same model, hereafter referred to as the Model. The Model's area under the ROC curve in the training and validation groups was 0.923 (95% CI 0.884-0.963) and 0.809 (95% CI 0.675-0.943), respectively. The optimal cutoff for the training group was 0.858, with a sensitivity of 0.801, a specificity of 0.973 and an accuracy of 0.837; when this cutoff value was taken, the validation group's sensitivity was 0.689, specificity was 0.800, and accuracy was 0.711. The Model demonstrated excellent performance in the calibration curve, Hosmer-Lemeshow test ( P>0.05), and DCA. Finally, based on the five predictors of the Model, a dynamic nomogram was created for clinicians to enter baseline clinical parameters for early identification of high-risk patients with abnormal bone mass. Conclusions:A dynamic nomogram for abnormal bone mass in ESRD patients is constructed with good predictive performance based on the prediction model, which can be used as a practical approach for the personalized early screening and auxiliary diagnosis of the potential risk factors and assist physicians in making a personalized diagnosis for patients.
8.G protein alpha inhibitory subunit 1/3 mediate Netrin-1-activated signaling and angiogenesis
Yujia YAO ; Jiajun LI ; Suyu WANG ; Keran LI
Chinese Journal of Ocular Fundus Diseases 2024;40(10):781-789
Objective:To observe the effect of G protein alpha inhibitory subunit (Gαi) 1 and Gαi3 on signal transduction and angiogenesis induced by Netrin-1 (NTN1) and explore the possible mechanisms.Methods:Twenty male C57BL/6J mice aged 6 to 8 weeks were randomly assigned to a control group and a diabetic group, with 10 mice in each group. Diabete group mice were induced by streptozotocin to establish diabetes model. 12 weeks after modeling, quantitative real-time polymerase chain reaction and Western blot were performed to detect the expression of Ntn1, Gαi1 and Gαi3 in diabetic retinas. Additionally, 35 male C57BL/6J mice aged 2 weeks were randomly stratified into three groups: a control group, an intravitreal injection of NTN1 group (NTN1 group), and a retinal endothelial cell-specific Gαi1/Gαi3 knockdown coupled with intravitreal NTN1 injection group (Gαi1/Gαi3 eKD+NTN1 group), with 15 mice in each of the normal control and NTN1 groups, and 5 mice in the Gαi1/Gαi3 eKD+NTN1 group. Isolectin B4 staining was performed to observe retinal neovascularization. In vitro, human umbilical vein endothelial cells (HUVEC) were divided into four groups: negative control lentiviral transfection group (shC group), negative control lentiviral transfection+NTN1 treatment group (shC+NTN1 group), Gαi1/Gαi3 knockdown group (shGαi1/Gαi3 group), and Gαi1/Gαi3 knockdown+NTN1 treatment group (shGαi1/Gαi3+NTN1 group). The effects of NTN1, Gαi1, and Gαi3 on HUVEC proliferation were assessed using the EdU assay. Transwell assays were conducted to determine the effects on HUVEC migration, and Matrigel assays were used to evaluate the effects on HUVEC tube formation. Protein kinase B (Akt), phosphorylated Akt (p-Akt), ribosomal protein S6 kinase (S6K), phosphorylated S6K (p-S6K), extracellular regulatory protein kinase (Erk1/2), phosphorylated Erk1/2 (p-Erk1/2) protein expression on HUVEC were detected by Western blot.Results:Compared with the control group, the relative expression levels of Ntn1, Gαi1, and Gαi3 mRNA and protein in the diabetic group retina were significantly increased, with statistically significant differences ( t=11.800, 9.298, 10.620, 7.503, 3.432, 8.037; P<0.000 1). Compared with the shC group, the relative expression levels of Gαi1 and Gαi3 mRNA and protein in the shGαi1/Gαi3 group in HUVEC were significantly reduced, showing statistically significant differences ( t=16.310, 16.300, 13.600, 9.068; P<0.000 1). HUVEC proliferation rate, migration number and lumen formation number: compared with shC group, shC+NTN1 group significantly increased, while shGαi1/Gαi3 group and shGαi1/Gαi3+NTN1 group significantly decreased, and the differences were statistically significant ( F=62.750, 49.830, 54.900; P<0.000 1). Compared with the control group, the relative expression levels of Gαi1 and Gαi3 mRNA and protein in retina were significantly decreased in the Gαi1/Gαi3 eKD group, showing statistically significant differences ( t=10.920, 13.460, 9.219, 10.500; P<0.000 1). Retinal neovasculogenesis area: compared with the normal control group, the area of retinal neovasculogenesis increased significantly in the NTN1 group, but decreased significantly in the Gαi1/Gαi3 eKD+NTN1 group, with statistical significance ( F=24.010, P<0.000 1). The protein expression of p-Akt relative to Akt, p-S6K relative to S6K, and p-Erk1/2 relative to Erk1/2: compared with shC group, the protein expression of shC+NTN1 group was significantly increased, while that of shGαi1/Gαi3 group and shGαi1/Gαi3+NTN1 group was significantly decreased, with statistical significance ( F=78.610, 144.400, 77.010; P<0.000 1). Conclusions:NTN1 induces Gαi1/Gαi3 to mediate activation of downstream Akt-mammalian target proteins of rapamycin and Erk1/2, thereby promoting angiogenesis in vivo and in vitro environments. Knocking down Gαi1/Gαi3 significantly reduces the NTN1-induced angiogenesis effect.
9.Targeted Inhibition of p21 Promotes the Growth of Breast Cancer Cells and Impairs the Tumor-Killing Effect of the Vaccinia Virus
Xiaoyuan JIA ; Yujia ZHAO ; Qiang LI ; Xiaming LU ; Xiaoyan WANG ; Hui WANG ; Ziyi SHI ; Yipeng XU ; Biao HUANG ; Fang HUANG ; Yigang WANG
Journal of Breast Cancer 2024;27(5):293-304
Purpose:
Vaccinia virus is widely used as an oncolytic agent for human cancer therapy, and several versions of vaccinia virus have demonstrated robust antitumor effects in breast cancer. Most vaccinia viruses are modified by thymidine kinase (TK) deletion. The function of the cyclin-dependent kinase inhibitor p21 in breast cancer remains controversial. We explored the impact of p21 gene knockdown (KD) on breast cancer cells and whether p21 KD interferes with the antitumor effect of TK-negative vaccinia virus.
Methods:
p21 KD MDA-MB-231 and p21 KD MCF-7 cells were prepared, and cell proliferation and migration rates were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and scratch healing assays. The tumor growth of xenografts originating from p21KD MDA-MB-231 cells and control cells was compared in a mouse model. The colony formation and sphere-forming abilities of p21 KD breast cancer cells were also determined using low-melting agarose and serum-free culture. The tumorkilling effect of the vaccinia virus was determined in breast cancer cells and mouse models using an MTT assay and tumor cell xenografts.
Results:
p21 KD increased the growth and migration of MDA-MB-231 and MCF-7 cells and promoted the cell growth of MDA-MB-231 cells in mice, while decreasing the colony formation and sphere formation abilities. Expression of TK was reduced in p21 KD MDAMB-231 cells. Oncolytic effects of both wild-type and TK-deleted vaccinia viruses were attenuated in p21KD MDA-MB-231 cells. The tumor-killing effect of TK-deleted vaccinia virus was also weakened in xenografted mice bearing p21 KD MDA-MB-231 cells.
Conclusion
Targeted inhibition of p21 accelerates the proliferation and migration of breast cancer cells and impairs the tumor-killing effect of vaccinia virus, suggesting that p21 levels in cancer cells interfere with vaccinia virus oncolytic therapy.
10.Effect of Icariin on Peripheral Blood Dendritic Cells and Th17/Treg Balance in Myocardial Remodeling Model of Dahl Salt-sensitive Rats
Qian LI ; Yan ZHOU ; Yujia CHEN ; Wen LI ; Huanzhen WANG ; Liancheng GUAN ; Yunzhi CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):46-55
ObjectiveTo investigate the effect of icariin (ICA)-mediated vitamin D system on peripheral blood dendritic cells (DCs) and helper T cells 17 (Th17)/regulatory T cells (Treg) balance in myocardial remodeling model of Dahl salt-sensitive rats. MethodFifty SPF Dahl salt-sensitive rats were divided into model group, vitamin D group (3×10-5 mg·kg-1·d-1), and high-, medium-, and low-dose ICA groups (120, 60, 30 mg·kg-1·d-1), and 10 Dahl salt-resistant rats were used as normal group. The myocardial remodeling model was established by feeding rats with a high-salt diet containing 8% NaCl. After six weeks of modeling, the normal group and the model group were given an equal volume of ultrapure water by gavage, and other groups were continuously administrated for six weeks. Cardiac echocardiography, hematoxylin-eosin (HE) staining, and Masson staining were used to observe the pathological changes in cardiac structure and fibrosis. The levels of serum 25(OH)D3, B-type N-terminal pro-brain natriuretic peptide (NT-ProBNP), interleukin (IL)-17, transforming growth factor (TGF)-β1, IL-12, and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA). The phenotype of peripheral blood DCs and the ratio of Th17/Treg cells of rats were detected by flow cytometry. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the mRNA and protein expressions of vitamin D receptor (VDR),1α-hydroxylase (CYP27B1), and 24-hydroxylase (CYP24A1) in peripheral blood DCs of rats. ResultCompared with the control group, the rats in the model group had pathological changes such as disordered arrangement of myocardial cells and cytoplasmic hypertrophy and swelling. Myocardial collagen fibers proliferated significantly, and the arrangement of myocardial fibers was disordered. The levels of serum 25(OH)D3 and IL-10 were significantly decreased, and the levels of serum IL-17, TGF-β1, IL-6, IL-12, and NT-ProBNP were significantly increased (P<0.05). The costimulatory molecules CD40, CD80, CD86, and MHC-Ⅱ were highly expressed in the peripheral blood DCs, and the expression of CD11 and CD11b was lower (P<0.05). The proportion of Th17 cells in the peripheral blood was significantly increased, and the proportion of Treg cells was decreased. The ratio of Th17/Treg was increased (P<0.05). The mRNA and protein expressions of CYP24A1 in peripheral blood DCs increased, and the mRNA and protein expressions of CYP27B1 and VDR decreased (P<0.05). Compared with the model group, the arrangement of myocardial fibers in each drug administration group was relatively regular, and the swelling of myocardial cells was significantly reduced. The pathological morphology of myocardial tissue was improved to varying degrees. The pathological changes in myocardial tissue were improved and alleviated to varying degrees. The drug could reduce the serum levels of NT-ProBNP, IL-17, TGF-β1, IL-6, and IL-12 and increase the level of serum 25(OH)D3 and IL-10 (P<0.05). The expression of costimulatory molecules CD40, CD80, CD86, and MHC-Ⅱ in the peripheral blood DCs of rats was decreased, and the expression of CD11 and CD11b molecules was increased (P<0.05). The drug could reduce the proportion of Th17 cells in peripheral blood and the ratio of Th17/Treg cells and increase the proportion of Treg cells (P<0.05). It could decrease the mRNA and protein expressions of CYP24A1 in peripheral blood DCs of rats and elevate the mRNA and protein expression of VDR and CYP27B1 (P<0.05). ConclusionICA can regulate the phenotype of peripheral blood DCs and the ratio of Th17/Treg cells by regulating the vitamin D system and play a role in improving myocardial remodeling from the perspective of immune balance.

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