1.Yishen Huashi Granules Protect Kidneys of db/db Mice via p38 MAPK Signaling Pathway
Kaidong ZHOU ; Sitong WANG ; Ge JIN ; Yanmo CAI ; Xin ZHOU ; Yunhua LIU ; Xinxue ZHANG ; Min ZHANG ; Zongjiang ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):58-68
ObjectiveTo explore the mechanism of Yishen Huashi granules in alleviating renal tubular epithelial cell injury and relieving diabetic kidney disease by regulating the mitogen-activated protein kinase (MAPK) signaling pathway. MethodsThe db/db mice of 12 weeks old were randomly assigned into model , dapagliflozin (1.6 mg·kg-1), and Yishen Huashi granules (4.7 g·kg-1), and db/m mice were used as the control group. The general conditions of mice were observed, and fasting blood glucose and 24-h urinary protein and albumin-to-creatinine ratio (ACR) were measured at weeks 0 and 12 of administration. After 12 weeks of treatment, the levels of serum creatinine (SCr), blood urea (UREA), triglycerides (TG), total cholesterol (TC), and low density lipoprotein (LDL) were measured. The pathological changes in the renal tissue were observed by hematoxylin-eosin (HE) staining, Periodic acid-Schiff (PAS) staining, Mallory staining, and transmission electron microscopy. Real-time PCR was employed to determine the mRNA levels of monocyte chemotactic protein-1 (MCP-1) and CC chemokine receptor-2 (CCR2) in the renal tissue of mice. The immunohistochemical assay was employed to examine the expression of p38, phospho-p38 (p-p38), MCP-1, and CCR2 in the renal tissue of mice. Western blotting was employed to measure the protein levels of p-p38, p38, MCP-1, and CCR2 in the renal tissue of mice.HK-2 cells cultured in vitro were grouped as follows: negative control, high glucose(30 mmol·L-1), Yishen Huashi granule-containing serum, and SB203580. After 48 h of cell culture in each group, RNA were extracted and the levels of MCP-1, and CCR2 mRNA were determined by Real-time PCR,proteins were extracted and the levels of p38, p-p38, MCP-1, and CCR2 were determined by Western blot. ResultsThe in vivo experiments showed that before treatment, other groups had higher body weight, blood glucose level, 24 h urinary protein, and ACR than the control group (P<0.05,P<0.01). After 12 weeks of treatment, compared with the model group, the Yishen Huashi granules group showed improved general conditions, a decreasing trend in body weight, lowered levels of blood glucose, 24-h urinary protein, and ACR (P<0.01), reduced SCr and UREA (P<0.01), and declined levels of TC, TG, and LDL (P<0.05,P<0.01). Compared with the model group, the Yishen Huashi granules group showed alleviated damage and interstitial fibrosis in the renal tissue as well as reductions in glomerular foot process fusion and basement membrane thickening. Moreover, the Yishen Huashi granules group showed down-regulated mRNA levels of MCP-1 and CCR2 (P<0.01), reduced positive expression of p-p38, MCP-1, and CCR2 (P<0.01), and down-regulated protein levels of p-p38/p38, MCP-1, and CCR2 (P<0.05) in the renal tissue. The cell experiment showed that compared with the high glucose group, the Yishen Huashi granule-containing serum group showcased down-regulated mRNA levels of MCP-1 and CCR2 (P<0.01) and down-regulated protein levels of p-p38/p38, MCP-1, and CCR2(P<0.05,P<0.01). ConclusionYishen Huashi granules can regulate glucose-lipid metabolism, reduce 24 h urinary protein and ACR, improve the renal function, alleviate the renal tubule injury caused by high glucose, and protect renal tubule epithelial cells in db/db mice by reducing MCP-1/CCR2 activation via the p38 MAPK signaling pathway.
2.In Vitro and in vivo Component Identification of Danshenyin Based on UPLC-Q-TOF-MS/MS
Sitong ZHANG ; Xianrun HU ; Wenkang LIU ; Jinchun LEI ; Xuemei CHENG ; Xiaojun WU ; Wansheng CHEN ; Manlin LI ; Changhong WANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):175-183
ObjectiveTo elucidate the chemical composition of Danshenyin and its blood components in rats after oral administration. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) coupled with PeakView 1.2 software was used to systematically characterize and identify the components of Danshenyin aqueous extract and its migratory components in rat blood after oral administration based on the retention time, quasi-molecular ion peaks, secondary fragmentation ions, and literature reports, and a preliminary compounds identification of Salviae Miltiorrhizae Radix et Rhizoma aqueous extract, the co-decoction of Santali Albi Lignum and Amomi Fructus was carried out to attribute the chemical constituents of the aqueous extract of Danshenyin. ResultsA total of 73 compounds, including 21 phenolic acids, 23 diterpenes, 6 flavonoids, 7 organic acids, 3 volatile oils and 13 others, were identified from the aqueous extract of Danshenyin. And 36 prototypes and 15 metabolites were identified in rat plasma, the major metabolic pathways included reduction, hydration, hydroxylation, demethylation, methylation, sulfation and others, these metabolites were mainly derived from tanshinones and salvianolic acids. ConclusionThe main blood components of the aqueous extract of Danshenyin are salvianolic acids and tanshinones, which may be the material basis of the efficacy. This study can provide reference for pharmacological research, quality control, and clinical application of Danshenyin.
3.Investigation on the current status of pharmaceutical care in medical institutions in China
Tingting ZHENG ; Yuqing ZHANG ; Sitong LIU ; Wei ZHANG ; Jin LU ; Qunhong SHEN ; Jiancun ZHEN
China Pharmacy 2025;36(9):1117-1122
OBJECTIVE To investigate the current status of pharmaceutical care in medical institutions in China, and provide experience and suggestions for better development of pharmaceutical care. METHODS Questionnaire survey was used to investigate the development of pharmaceutical care in medical institutions in 31 provinces (autonomous regions and municipalities directly) in March 2023, and descriptive analysis and binary logistic regression analysis on the influencing factors of pharmaceutical care were conducted. RESULTS A total of 1 368 questionnaires were sent out, and 1 304 valid questionnaires were collected with the effective recovery rate of 95.32%. Pharmaceutical care was carried out in 671 medical institutions (51.46%), and the rates of pharmaceutical care in tertiary, secondary, primary and other medical institutions were 74.79%, 27.97% and 7.35%, respectively. The average number of patients receiving pharmaceutical care was 2 638.96 per year, and there were 8.33 pharmacists in each medical institution to carry out pharmaceutical care, among which 93.68% were clinical pharmacists. The main departments covered by pharmaceutical care services included respiratory and critical care medicine, cardiology, intensive care unit, endocrinology, oncology, gastroenterology, obstetrics and gynecology and other departments. There were only 48 medical institutions (7.15%) with additional compensation for pharmaceutical care services. The main experiences of developing pharmaceutical care were to pay attention to talent cultivation and discipline construction, but the main difficulties were serious shortage of staff and qualified talents, low compensation level and low enthusiasm. Grade of medical institutions, the number of pharmacists engaged in clinical pharmacy, the number of qualified clinical pharmacists and the degree of information in the pharmacy department were the main influencing factors for carrying out pharmaceutical care (P<0.05). CONCLUSIONS In recent years, pharmaceutical care in Chinese medical institutions has made certain progress, while that of primary medical institutions, secondary medical institutions and other medical institutions should be improved. In the future, it is still necessary to further enhance the implementation of pharmaceutical care, promote personnel training, and attach importance to demonstrating the value of pharmaceutical care, thereby promoting the sustainable and high- quality development of pharmaceutical care.
4.Effect of Shenkang Injection on Podocyte Apoptosis and GRP78/CHOP Signaling Pathway in db/db Mice with Diabetic Kidney Disease Based on Endoplasmic Reticulum Stress
Yanmo CAI ; Sitong WANG ; Xin ZHOU ; Ge JIN ; Kaidong ZHOU ; Yunhua LIU ; Fengfeng ZHANG ; Xinxue ZHANG ; Zongjiang ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):81-90
ObjectiveTo investigate the mechanism of Shenkang injection in delaying diabetic kidney disease by regulating endoplasmic reticulum stress and attenuating podocyte apoptosis through the Glucose regulated protein 78 ( GRP78 ) / transcription factor C / EBP homologous protein ( CHOP ) signaling pathway (GRP78/CHOP) signaling pathway. MethodsFor the animal experiment, 10 12-week-old db/m mice were selected as a normal group, and 30 12-week-old db/db mice were randomly divided into a model group, a Shenkang injection group (15.6 mL·kg-1), and a dapagliflozin group (1.6 mg·kg-1). To observe the general condition of mice, fasting blood glucose, urinary albumin/urine creatinine (ACR), and 24 h urine protein quantification were detected in each group before drug administration. After 12 weeks of drug treatment, mice were tested for fasting blood glucose, total cholesterol (TC), triglyceride (TG), low-density cholesterol (LDL), ACR, 24 h urine protein quantification, blood creatinine (SCr), and blood urea (UREA). Hematoxylin-eosin (HE) staining, periodic acid-Schiff (PAS) staining, and transmission electron microscopy were used to observe the pathologic morphology in renal tissue. Immunohistochemistry was used to detect the expressions of nephroprotective marker protein (Nephrin), glucose-regulated protein 78 (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in renal tissue. Western blot was used to detect the expressions of GRP78, CHOP, Bcl-2, Bax, and Nephrin proteins, and Real-time polymerase chain reaction (Real-time PCR) was employed to detect the expressions of Nephrin, GRP78, CHOP, Bcl-2, and Bax mRNAs in renal tissue. ResultsBefore drug administration, compared with those in the normal group, the body mass of db/db mice was significantly increased, and blood glucose, 24 h urine protein quantification, and ACR were significantly elevated in the Shenkang injection group and Dapagliflozin group (P<0.01). After 12 weeks of administration, compared with those in the model group, the general state of mice in the Shenkang injection group was significantly improved, and the body mass was decreased. The blood glucose was significantly reduced (P<0.01), and blood lipids TC, TG, and LDL were significantly decreased (P<0.05, P<0.01). The 24 h urine protein quantification and ACR were significantly decreased (P<0.05), and SCr and UREA were significantly reduced (P<0.01). Compared with those of the model group, the pathologic results of the Shenkang injection group showed that proliferation of mesangial cells, reduction of inflammatory cell infiltration, and alleviation of renal tubular vacuolization and podocyte damage were observed in renal tissue of mice. Electron microscopy showed that fusion of the pedicle protruding and thickening of the basement membrane were reduced. Immunohistochemistry results showed that the expressions of GRP78, CHOP, and Bax proteins were significantly reduced (P<0.01), and the expressions of Nephrin and Bcl-2 proteins were significantly increased (P<0.01) in renal tissue of the Shenkang injection group. Western blot results showed that the expressions of Nephrin and Bcl-2 in the Shenkang injection group were significantly increased (P<0.05, P<0.01), and the expressions of GRP78, CHOP, and Bax proteins were significantly decreased (P<0.05, P<0.01). Real-time PCR results showed that the expressions of GRP78, CHOP, and Bax mRNAs were down regulated in the Shenkang injection group (P<0.01), and the expressions of Nephrin and Bcl-2 mRNAs were up regulated (P<0.01). ConclusionShenkang injection inhibits endoplasmic reticulum stress response and podocyte apoptosis by regulating the GRP78/CHOP signaling pathway, which in turn ensures the integrity of glomerular filtration barrier, reduces the occurrence of proteinuria, improves renal function, and thus delays the progression of diabetic kidney disease.
5.Effect of Shenkang Injection on Podocyte Apoptosis and GRP78/CHOP Signaling Pathway in db/db Mice with Diabetic Kidney Disease Based on Endoplasmic Reticulum Stress
Yanmo CAI ; Sitong WANG ; Xin ZHOU ; Ge JIN ; Kaidong ZHOU ; Yunhua LIU ; Fengfeng ZHANG ; Xinxue ZHANG ; Zongjiang ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):81-90
ObjectiveTo investigate the mechanism of Shenkang injection in delaying diabetic kidney disease by regulating endoplasmic reticulum stress and attenuating podocyte apoptosis through the Glucose regulated protein 78 ( GRP78 ) / transcription factor C / EBP homologous protein ( CHOP ) signaling pathway (GRP78/CHOP) signaling pathway. MethodsFor the animal experiment, 10 12-week-old db/m mice were selected as a normal group, and 30 12-week-old db/db mice were randomly divided into a model group, a Shenkang injection group (15.6 mL·kg-1), and a dapagliflozin group (1.6 mg·kg-1). To observe the general condition of mice, fasting blood glucose, urinary albumin/urine creatinine (ACR), and 24 h urine protein quantification were detected in each group before drug administration. After 12 weeks of drug treatment, mice were tested for fasting blood glucose, total cholesterol (TC), triglyceride (TG), low-density cholesterol (LDL), ACR, 24 h urine protein quantification, blood creatinine (SCr), and blood urea (UREA). Hematoxylin-eosin (HE) staining, periodic acid-Schiff (PAS) staining, and transmission electron microscopy were used to observe the pathologic morphology in renal tissue. Immunohistochemistry was used to detect the expressions of nephroprotective marker protein (Nephrin), glucose-regulated protein 78 (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax) in renal tissue. Western blot was used to detect the expressions of GRP78, CHOP, Bcl-2, Bax, and Nephrin proteins, and Real-time polymerase chain reaction (Real-time PCR) was employed to detect the expressions of Nephrin, GRP78, CHOP, Bcl-2, and Bax mRNAs in renal tissue. ResultsBefore drug administration, compared with those in the normal group, the body mass of db/db mice was significantly increased, and blood glucose, 24 h urine protein quantification, and ACR were significantly elevated in the Shenkang injection group and Dapagliflozin group (P<0.01). After 12 weeks of administration, compared with those in the model group, the general state of mice in the Shenkang injection group was significantly improved, and the body mass was decreased. The blood glucose was significantly reduced (P<0.01), and blood lipids TC, TG, and LDL were significantly decreased (P<0.05, P<0.01). The 24 h urine protein quantification and ACR were significantly decreased (P<0.05), and SCr and UREA were significantly reduced (P<0.01). Compared with those of the model group, the pathologic results of the Shenkang injection group showed that proliferation of mesangial cells, reduction of inflammatory cell infiltration, and alleviation of renal tubular vacuolization and podocyte damage were observed in renal tissue of mice. Electron microscopy showed that fusion of the pedicle protruding and thickening of the basement membrane were reduced. Immunohistochemistry results showed that the expressions of GRP78, CHOP, and Bax proteins were significantly reduced (P<0.01), and the expressions of Nephrin and Bcl-2 proteins were significantly increased (P<0.01) in renal tissue of the Shenkang injection group. Western blot results showed that the expressions of Nephrin and Bcl-2 in the Shenkang injection group were significantly increased (P<0.05, P<0.01), and the expressions of GRP78, CHOP, and Bax proteins were significantly decreased (P<0.05, P<0.01). Real-time PCR results showed that the expressions of GRP78, CHOP, and Bax mRNAs were down regulated in the Shenkang injection group (P<0.01), and the expressions of Nephrin and Bcl-2 mRNAs were up regulated (P<0.01). ConclusionShenkang injection inhibits endoplasmic reticulum stress response and podocyte apoptosis by regulating the GRP78/CHOP signaling pathway, which in turn ensures the integrity of glomerular filtration barrier, reduces the occurrence of proteinuria, improves renal function, and thus delays the progression of diabetic kidney disease.
6.Influence of SH2B1 rs7359397 polymorphism on hepatic fibrosis in patients with metabolic dysfunction-associated steatotic liver disease aged ≥65 years in Beijing, China
Sitong CHEN ; Shuang ZHANG ; Jinhan ZHAO ; Xiaodie WEI ; Yaning LI ; Lixia QIU ; Jing ZHANG
Journal of Clinical Hepatology 2025;41(11):2286-2293
ObjectiveTo investigate the association of SH2B1 rs7359397 (C>T) polymorphism with the progression to hepatic fibrosis in the elderly patients with metabolic dysfunction-associated steatotic liver disease (MASLD) in Beijing, China, and to provide an important genetic basis for the precise subtyping, prognostic evaluation, and individualized treatment of elderly MASLD patients in China. MethodsA total of 505 elderly patients (aged ≥65 years) who participated in regular physical examination in Mentougou Kuangshan Hospital of Beijing Jingmei Group General Hospital from November 2020 to September 2021 and were diagnosed with MASLD by abdominal ultrasound were enrolled as MASLD group, and 381 elderly population who underwent physical examination in the same community hospital during the same period of time and were not found to have MASLD by abdominal ultrasound were enrolled as control group. FibroScan was used to measure liver fat content and determine fibrosis stage. The 96-well microfluidic chip technique was used to identify SH2B1 rs7359397 polymorphism. The independent-samples t test was used for comparison of normally distributed continuous data between the two groups, and the chi-square test or the adjusted chi-square test was used for comparison of categorical data between the two groups. Univariate and multivariate Logistic regression analyses were used to identify the independent predictive factors for MASLD and its comorbidities. ResultsCompared with the control group, the MASLD group had a significantly younger age and significantly higher levels of waist circumference, hip circumference, waist-hip ratio, body mass index (BMI), alanine aminotransferase, aspartate aminotransferase, triglyceride, platelet count, and fibrosis-4 (FIB-4) index, as well as a significantly lower level of high-density lipoprotein cholesterol (all P<0.05). Among the 381 patients in the control group, 264 (69.29%) had genotype CC and 117 (30.71%) had genotype CT+TT, while among the 505 patients in the MASLD group, 317 (62.77%) had genotype CC and 188 (37.23%) had genotype CT+TT, suggesting that the MASLD group had a significantly higher proportion of patients with genotype CT+TT compared with the control group (χ2=4.09, P=0.043). In the MASLD group, compared with the genotype CC group, the genotype CT+TT group had a significantly lower proportion of patients with FIB-4 ≥2 or atherosclerotic cardiovascular diseases (P<0.05). The multivariate Logistic regression analysis showed that after adjustment for age, sex, and BMI, carrying T allele was a protective factor against progressive hepatic fibrosis (odds ratio [OR]=0.481, 95% confidence interval [CI]: 0.249 — 0.929, P=0.029). In the subgroups of comorbidities with hypertension, metabolic syndrome, and obesity, genotype CT+TT was associated with a significant reduction in the risk of progressive hepatic fibrosis (hypertension: OR=0.27, 95%CI:0.09 — 0.77, P=0.014; metabolic syndrome: OR=0.30, 95%CI: 0.11 — 0.79, P=0.015; obesity: OR=0.11, 95%CI: 0.03 — 0.48, P=0.003). After adjustment for age, sex, and BMI, in the patients with MASLD, the patients with genotype CT+TT had a significant reduction in the prevalence rate of atherosclerotic cardiovascular diseases compared with those with genotype CC (OR=0.506, 95%CI:0.336 — 0.761, P=0.001). ConclusionSH2B1 rs7359397 (C>T) polymorphism is associated with the reduction in the risk of hepatic fibrosis and atherosclerotic cardiovascular diseases in MASLD patients.
7.Reactive plasmacytosis caused by methimazole in patients with Graves'disease:One case report and literature review
Shimeng LI ; Xin QI ; Sitong LIN ; Xiangwen SAN ; Ling JIN ; Sitong ZHANG
Journal of Jilin University(Medicine Edition) 2024;50(5):1414-1419
Objective:To discuss the clinical manifestations and laboratory examination results of agranulocytosis and reactive plasmacytosis(RP)in the patient with Graves'disease(GD)after treated with methimazole(MMI),and to provide the basis for the clinicians to differentiate RP from multiple myeloma(MM).Methods:The clinical manifestations,laboratory examinations,diagnosis and treatment processes of one patient with GD agranulocytosis complicated with RP were analyzed,and the related literatures were reviewed.Results:The patient had a history of GD and abdominal infection.Upon admission,a complete blood count revealed a significant decrease in white blood cell count accompanied by neutropenia,and a smear re-examination showed suspicious plasma cells.The bone marrow cytology examination results showed the percentage of bone marrow plasma cells was 33%,and the percentage of plasma cells in peripheral blood was 4%;the serum immunoglobulin results showed polyclonal hyperplasia;the serum immunofixation electrophoresis results were negative;the flow cytometry analysis results indicated the immunophenotype of the plasma cells was normal.Based on the medical history and laboratory results,MM was largely excluded,supporting the diagnosis of RP.Neutropenia was considered to be related to medication,so MMI was discontinued,granulocyte colony-stimulating factor was administered to increase the number of white blood cells,and specialized GD treatment was conducted after controlling the abdominal infection.The patient had a good prognosis,and his blood count was normal upon re-examination 6 months later.Conclusion:Agranulocytosis complicated with RP in the GD patients is clinically rare.Serum immunofixation electrophoresis,blood cell morphology,and cell immunophenotype analysis are helpful for the accurate diagnosis.After actively treating the primary disease causing RP,the patient's prognosis is favorable.
8.Study on efficacy of arthroscopic simple repair technique and enhanced repair technique in treating chronic ankle instability
Bao LI ; Xinqi LIN ; Haokun LI ; Zhiqiang ZHANG ; Sitong LI ; Bowen FANG ; Fangfang WU ; Xinwei LIU
Chongqing Medicine 2024;53(12):1784-1788,1792
Objective To investigate the clinical efficacy of arthroscopic simple repair technique and en-hanced repair technique in the treatment of chronic lateral ankle instability(CLAI).Methods Forty-one cases of CLAI treated in the General Hospital of Northern Theater Command of PLA from January 2018 to January 2019 were selected and conducted arthroscopic lateral ankle ligament repair or enhanced repair treatment.The follow up data in 39 cases were complete.Among them,18 cases conducted the anterior talofibular ligament repair with wire anchor under arthroscopy and were included in the repair group,while 21 cases conducted the arthroscopic enhanced repair with linear anchors and knots free anchors and were included in the enhanced re-pair group.The postoperative follow up lasted for 12 months.The VAS score and AOFAS ankle-hind foot score before operation and in postoperative,3,6,12 months were compared between the two groups.The effi-cacies of treating CLAI by the two operation modes were evaluated.Results The preoperative VAS score and AOFAS ankle-hind foot score had no statistical differences between the two groups(P>0.05).The VAS score at postoperative time points had no statistical difference between the enhanced repair group and repair group(P>0.05).The AOFAS ankle-hind foot score in the enhanced repair group was higher than that in the repair group,and the difference was statistically significant(P<0.05).No serious complications occurred in the grouped patients during the follow up period.Conclusion In terms of pain and ankle function,the effect of arthroscopic enhanced repair in treating CLAI is better than that of simple repair.
9.Effects of Polydatin on Proliferation and Apoptosis of Pulmonary Artery Smooth Muscle Cells in Neonatal Rats with Hypoxic Pulmonary Hypertension by Regulating YAP1/TAZ Signaling Pathway
Wenguang LIU ; Sitong LIU ; Weiwei ZHANG
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 2024;53(1):68-74
Objective To investigate the effects of polydatin(PD)on the proliferation and apoptosis of pulmonary artery smooth muscle cells(PASMCs)in hypoxic pulmonary hypertension(HPH)neonatal rats,and its mechanism of action.Methods Neonatal rats were randomly separated into six groups:control group,model group,low dose PD group,medium dose PD group,high dose PD group,and high dose PD+Hippo pathway inhibitor(high dose PD+XMU-MP-1)group,with 10 rats in each group.After 2 weeks of hypoxia treatment,the right ventricular systolic blood pressure(RVSP)and right ventricular hypertro-phy index(RVHI)of rats in each group were measured.Hematoxylin-eosin(HE)staining was applied to observe pathological changes in lung tissue,and the percentage of pulmonary artery wall thickness to total thickness(WT)and the percentage of wall area to total area(WA)were calculated.Neonatal rat PASMCs were separated from each group,which were divided into NC group,hypoxia group,low dose PD group,medium dose PD group,high dose PD group,and high dose PD+XMU-MP-1 group.Cell counting kit 8(CCK-8)and 5-ethynyl-2'-deoxyuridine(EdU)were applied to detect cell proliferation.Flow cytometry was applied to detect cell apoptosis.Western blot was applied to detect the expression of Yes-associated protein 1(YAP1),tran-scriptional coactivator with PDZ-binding motif(TAZ),mammalian sterile 20-like kinase 1(MST1),B-cell lymphoma 2(Bcl-2),and Bcl-2 associated protein(Bax)in lung tissue and PASMCs.Results Compared with the control group,the pulmonary artery wall in the model group was significantly thickened,lumen was narrowed,and protein expressions of RVSP,RVHI,WT%,WA%,YAP1,MST1 and TAZ were significantly increased(all P<0.05).Compared with the model group,pulmonary artery thickening and lumen enlargement were observed in the low,medium and high dose PD groups,and the protein expressions of RVSP,RVHI,WT%,WA%,YAP1,MST1 and TAZ were significantly decreased,which showed a dose-dependent relationship(all P<0.05).The effect could be reversed by XMU-MP-1.Compared with the NC group,the cell A450nm value,EdU positive rate,the protein expression of YAP1,MST1,TAZ and Bcl-2 in the hydropoxia group were significantly increased.The apoptosis rate and the expression of Bax protein were obviously reduced(all P<0.05).Compared with the hypoxia group,the cell A450nm value,EdU positive rate,the protein expression of YAP1,MST1,TAZ and Bcl-2 in the low,medium and high dose PD groups were obviously reduced.The apoptosis rate and the expression of Bax were significantly increased,which showed a dose-depend-ent relationship(all P<0.05).The effect could be reversed by XMU-MP-1.Conclusion PD may inhibit the proliferation of PASMCs in HPH neonatal rats and promote apoptosis by inhibiting YAP1/TAZ signaling pathway.
10.Mechanism of Total Saponin of Astragali Radix and Total Alkaloids of Nelumbinis Folium Against Hyperlipidemia Based on PPARγ/LXRα/ABCG1 Signaling Pathway
Yufang ZHANG ; Jian CHEN ; Yixin ZHANG ; Sitong LI ; Yafen WANG ; Yongqi ZHANG ; Cheng SHI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(13):37-44
ObjectiveTo observe the effect of the combination of total saponin of Astragali Radix-total alkaloids of Nelumbinis Folium on reversal cholesterol transport (RCT) in hyperlipidemia rats, and to discuss its mechanism. MethodSixty SD rats were randomly divided into control group, high-fat diet group, total saponin of Astragali Radix-total alkaloids of Nelumbinis Folium low (17 mg·kg-1+40 mg·kg-1), middle (34 mg·kg-1+80 mg·kg-1), high dose (68 mg·kg-1+160 mg·kg-1) groups and simvastatin (2.1 mg·kg-1) group, with 10 mice in each group. The Hyperlipidemia model was duplicated by feeding rats with a high-fat diet for 6 weeks. From the 3rd week, except for the control group and the high-fat diet group given distilled water, other groups were given corresponding drugs intragastric treatment for 4 weeks. The changes in blood lipid and liver function of rats were detected by an automatic biochemical analyzer. Hematoxylin-eosin (HE) and oil red O staining were used to observe the pathological morphological changes and steatosis of rat liver tissue. The contents of total cholesterol (TC) and total bile acid (TBA) in rat liver tissue and feces were determined by a semi-automatic biochemical analyzer. The mRNA and protein expression levels of peroxisome proliferators-activated receptors γ (PPARγ), liver X receptors α (LXRα), ATP-binding cassette transporter G1 (ABCG1) and cholesterol 7α-hydroxylase (CYP7A1) in rat liver tissue were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot. ResultCompared with the control group, the contents or activities of TC, triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), TBA, aspartate aminotransferase (AST), alanine aminotransferase (ALT) in serum were significantly increased (P<0.01), and the contents of high-density lipoprotein cholesterol (HDL-C) in the high-fat diet group were significantly decreased (P<0.01). The hepatocyte was clearly swollen like ballooning degeneration, with a lot of fat vacuoles and red fat droplets. The contents of TC and TBA in liver tissue and feces were significantly increased (P<0.01), and the mRNA and protein expression levels of PPARγ, LXRα, ABCG1, and CYP7A1 in liver tissue were significantly decreased (P<0.01). Compared with the high-fat diet group, the contents or activities of TC, TG, LDL-C, TBA, AST, and ALT in the serum of rats in administered groups were significantly decreased (P<0.01), while the content of HDL-C was significantly increased (P<0.01). Hepatocyte swelling was significantly reduced, and the ballooning degeneration, fat vacuoles, and red lipid droplets in liver tissue were significantly decreased. The contents of TC and TBA in liver tissue were significantly decreased (P<0.05, P<0.01), and the contents of TC and TBA in feces were significantly increased (P<0.05, P<0.01). The mRNA and protein expression levels of PPARγ, LXRα, ABCG1, and CYP7A1 in liver tissue were significantly increased (P<0.05, P<0.01). ConclusionTotal saponin of Astragali Radix-total alkaloids of Nelumbinis Folium has a positive effect on the prevention and treatment of hyperlipidemia rats, and its mechanism may be related to the activation of PPARγ/LXRα/ABCG1 signaling pathway and regulation of RCT.

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