1.Expert Consensus on Clinical Application of Pingxuan Capsules
Yuer HU ; Yanming XIE ; Yaming LIN ; Yuanqi ZHAO ; Yihuai ZOU ; Mingquan LI ; Xiaoming SHEN ; Wei PENG ; Changkuan FU ; Yuanyuan LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):201-210
As a patented characteristic medicine of Yi ethnic minority, Pingxuan capsules have the effects of nourishing the liver and kidney, pacifying the liver, and subduing Yang. With the main indications of dizziness, headache, palpitations, tinnitus, insomnia, dreaminess, waist and knee soreness caused by liver-kidney deficiency and liver Yang upward disturbance, Pingxuan capsules are widely used in the treatment of posterior circulation ischemic vertigo, vestibular migraine, benign paroxysmal positional vertigo. However, the current knowledge is limited regarding the efficacy, syndrome differentiation, and safety of this medicine. On the basis of summarizing the experience of clinicians and the existing evidence, this study invites clinical experts of traditional Chinese and Western medicine, pharmaceutical experts, and methodological experts from relevant fields across China to conduct evidence-based evaluation of Pingxuan capsules. The evaluation follows the Specifications for the Development of Clinical Expert Consensus on Chinese Patent Medicines issued by the Standardization Office of the China Association of Chinese Medicine, and reaches 5 recommendations and 16 consensus suggestions. The consensus clarifies the clinical applications, efficacy, dose, course of treatment, combination of medicines, precautions, and contraindications of Pingxuan capsules in the treatment of vertigo and explains the safety of clinical application. This consensus is applicable to clinicians (traditional Chinese medicine, Western medicine, and integrated traditional Chinese and Western medicine) and pharmacists in tertiary hospitals, secondary hospitals, and community-level medical and health institutions across China, providing a reference for the rational use of Pingxuan capsules in the treatment of vertigo. It is hoped that the promotion of this consensus can facilitate the rational use of drugs in clinical practice, reduce the risk of drug use, and give full play to the advantages of Pingxuan capsules in the treatment of vertigo diseases. This consensus has been reviewed and published by the China Association of Chinese Medicine, with the number GS/CACM330-2023.
2.Analysis of Major Syndromes and Their Typical Related Symptoms and Signs in 135 Patients with Metabolic Syndrome:A Clinical Study Based on Syndrome Element Differentiation and Latent Class Analysis
Tong WANG ; Mingqian JIANG ; Lifen MI ; Shanyi SHEN ; Shujie XIA ; Candong LI
Journal of Traditional Chinese Medicine 2025;66(4):376-381
ObjectiveTo explore the typical syndromes and their characteristic of symptoms and signs with high diagnostic value in patients with metabolic syndrome (MS). MethodsTraditional Chinese medicine (TCM) diagnostic information was collected from 135 MS patients. Syndrome element differentiation and latent class analysis (LCA) were applied to identify the major TCM syndromes in MS patients. Symptoms were analyzed based on the differentiated syndromes, and a binary logistic regression model was constructed to determine symptoms and signs with high diagnostic value. ResultsA total of 135 MS patients were included, involving 163 symptoms and signs with a total frequency of 1749; twenty-three syndrome elements were extracted, 367 times frequency in total, among which 8 syndrome elements occurred ≥10 times with 323 frequencies (88.01% of the total). These included location-related elements such as kidney (48 times), spleen (14 times), and stomach (14 times), and nature-related elements such as phlegm (71 times), yin deficiency (64 times), dampness (57 times), heat (42 times), and qi deficiency (13 times). Based on LCA, the 135 patients were categorized into two groups distinguished by the syndrome elements of dampness and phlegm, forming the "phlegm-dampness syndrome" as the major syndrome type. Nine high-frequency symptoms and signs associated with the phlegm-dampness syndrome were identified,i.e. obesity (39 times), greasy coating (38 times), slippery pulse (33 times), white coating (31 times), preference for fatty and heavy foods (30 times), excessive urination (30 times), fatigue and lack of strength (29 times), wiry pulse (25 times), and dark red tongue (25 times). A binary logistic regression model was constructed combining these nine symptoms and signs with the LCA classification results, ultimately identifying obesity, greasy coating, fatigue and lack of strength, and white coating as independent factors associated with the phlegm-dampness syndrome in MS patients (P<0.05). ConclusionThe major TCM syndrome in MS patients is phlegm-dampness syndrome, and obesity, greasy coating, fatigue and lack of strength, and white coating are the typical symptoms and signs for diagnosing phlegm-dampness syndrome in MS patients.
3.Efficacy Mechanism of Xianlian Jiedu Prescription Against Colorectal Cancer Recurrence vias Regulating Angiogenesis
Yanru XU ; Lihuiping TAO ; Jingyang QIAN ; Weixing SHEN ; Jiani TAN ; Chengtao YU ; Minmin FAN ; Changliang XU ; Yueyang LAI ; Liu LI ; Dongdong SUN ; Haibo CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):79-87
ObjectiveTo explore effect of Xianlian Jiedu prescription on the recurrence of colorectal cancer (CRC) and investigate the related mechanisms. MethodsA postoperative recurrence model was established in 25 Balb/c mice by injecting CT26 cells subcutaneously into the armpit, followed by surgical removal of 99% of the subcutaneous tumor. The mice were randomly divided into model group, low-dose Xianlian Jiedu prescription (XLJDP-L) group (6.45 g·kg-1·d-1), medium-dose Xianlian Jiedu prescription (XLJDP-M) group (12.9 g·kg-1·d-1), high-dose Xianlian Jiedu prescription (XLJDP-H) group (25.8 g·kg-1·d-1), and 5-fluorouracil (5-FU) group (1×10-3 g·kg-1·d-1). The mice were euthanized after 14 days of continuous intervention, and recurrent tumor tissue was harvested. Hematoxylin and eosin (HE) staining was used to observe pathological and morphological changes in the recurrent tumor tissue. Immunohistochemistry (IHC) was employed to assess the expression of proliferating cell nuclear antigen (Ki67), vascular endothelial growth factor (VEGF), and platelet-endothelial cell adhesion molecule (CD31) in recurrent tumor tissue. The Western blot was used to detect the protein expression levels of angiopoietin-2 (ANG-2), VEGF, phosphorylated-protein kinase B (p-Akt), protein kinase B (Akt), phosphorylated-phosphatidylinositol 3-kinase (p-PI3K), and phosphatidylinositol 3-kinase (PI3K) in recurrent tumor tissue. ResultsBefore treatment, there were no statistical differences in tumor volume, tumor weight, and body mass among the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group compared to the model group, indicating model stability. After treatment, compared with those in the model group, the tumor volume and tumor weight in the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly reduced (P<0.01), showing dose dependency. Meanwhile, there were no significant differences in body weight among the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group compared to the model group. HE staining showed that compared with that in the model group, tumor tissue in the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group had loosely arranged cells, increased intercellular spaces, small and shriveled nuclei, light staining, fewer mitotic figures and atypical nuclei, and increased necrotic areas. IHC showed that compared with those of the model group, the positive rates of Ki67, VEGF, and CD31 in the recurrent tumor tissue of the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly reduced (P<0.01) in a dose-dependent manner. Western blot results showed that compared with those of the model group, the protein expression levels of ANG-2 and VEGF in the recurrent tumor tissue of the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly downregulated (P<0.05, P<0.01), and the p-Akt/Akt and p-PI3K/PI3K ratios were significantly decreased in a dose-dependent manner (P<0.05, P<0.01). ConclusionXianlian Jiedu prescription significantly inhibits the recurrence of CRC in mice after subcutaneous tumor surgery. The mechanism may involve regulating the PI3K/Akt pathway and downregulating key angiogenic proteins such as ANG-2, VEGF, and CD31.
4.Clinical observation of doxycycline in the treatment of macrolide-unresponsive Mycoplasma pneumoniae pneumonia in children under 8 years of age
Shanshan LI ; Jishan ZHENG ; Yanyan CHEN ; Longhui SHEN
China Pharmacy 2025;36(4):464-468
OBJECTIVE To investigate the efficacy and safety of doxycycline in the treatment of macrolide-unresponsive Mycoplasma pneumoniae pneumonia (MUMPP) in children under 8 years of age. METHODS The medical records of children with MUMPP admitted to the Women and Children’s Hospital Affiliated to Ningbo University were collected from January 1st, 2023 to December 31st, 2023. They were divided into doxycycline group (44 cases), doxycycline combined with methylprednisolone group (35 cases), and azithromycin combined with methylprednisolone group (35 cases) according to the treatment methods. Doxycycline group was given Doxycycline hyclate enteric-coated capsules; doxycycline combined with methylprednisolone group was given Doxycycline hyclate enteric-coated capsules and Methylprednisolone sodium succinate for injection; azithromycin combined with methylprednisolone group was given Azithromycin for injection and Methylprednisolone sodium succinate for injection. Treatment courses of 3 groups lasted for 10 d. The fever reduction rate, the time of fever reduction and improvement rate of lung infection absorption were compared among the three groups. The occurrence of adverse drug reactions was recorded during their hospitalization and followed up within 5 months after discharge. RESULTS The fever reduction rats 48, 72 h after treatment and improvement rate of lung infection absorption in doxycycline group and doxycycline combined with methylprednisolone group were significantly higher than azithromycin combined with methylprednisolone group; the time of fever reduction was significantly shorter than azithromycin combined with methylprednisolone group (P<0.05); there was no statistical significance in the difference between the doxycycline group and the doxycycline combined with methylprednisolone group (P>0.05). There was no statistical significance in the incidence of rash, vomiting, abdominal pain, diarrhea, and elevated transaminases among the three groups during hospitalization and within 5 months after discharge (P>0.05). None of the children treated with doxycycline suffered from tooth discoloration or enamel hypoplasia. CONCLUSIONS Doxycycline has good efficacy and safety in therapy of MUMPP in children under 8 years of age; adjunctive coadministration of low-dose glucocorticoids does not necessarily result in significant additional efficacy.
5.The Effect of Zhiqiao Gancao Decoction (枳壳甘草汤) on Intervertebral Disc Nucleus Pulposus Cell Apoptosis and the Hippo-YAP/TAZ Signaling Pathway in Tail Intervertebral Disc Degeneration Model Rats
Zaishi ZHU ; Zeling HUANG ; Junming CHEN ; Bo XU ; Binjie LU ; Hua CHEN ; Xingxing DUAN ; Yuwei LI ; Xiaofeng SHEN
Journal of Traditional Chinese Medicine 2025;66(5):509-517
ObjectiveTo investigate the possible mechanism by which Zhiqiao Gancao Decoction (枳壳甘草汤, ZGD) delays intervertebral disc degeneration (IDD) based on the Hippo-yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ) signaling pathway. MethodsA total of 50 SD rats were randomly divided into sham surgery group, model group, low-dose ZGD group, high-dose ZGD group, and high-dose ZGD + inhibitor group, with 10 rats in each group. In the sham surgery group, the rats were pierced in the skin and muscle at the Co6/7/8 segments of the tail with a 21G needle (depth approximately 2 mm) without damaging the intervertebral disc. In the other groups, rats were injected with a 21G needle at the Co6/7/8 segments of the tail to establish an IDD model by piercing the tail intervertebral disc 5 mm. One week after modeling, rats in the low-dose and high-dose ZGD groups were given 6.24 and 12.24 g/(kg·d) of the decoction via gastric gavage, respectively. The high-dose ZGD + inhibitor group was given 12.24 g/(kg·d) of the decoction and an intraperitoneal injection of YAP/TAZ inhibitor Verteporfin 10 mg/kg. The sham surgery and model groups were given 5 ml/(kg·d) of normal saline via gavage. The gavage was given once a day, and the intraperitoneal injection was given every other day. After 4 weeks of continuous intervention, the pathological changes of the tail intervertebral discs were observed using HE staining, Oil Red O-Green staining, and Toluidine Blue staining. Immunohistochemistry was used to detect the expression of aggrecan and MMP3 in the nucleus pulposus. TUNEL fluorescence staining was performed to detect apoptosis in the nucleus pulposus, and the apoptosis rate was calculated. Western blot was used to detect the Hippo-YAP/TAZ signaling pathway, including YAP, phosphorylated YAP (p-YAP), phosphorylated MST1/2 (p-MST1/2), phosphorylated TAZ (p-TAZ) and apoptosis-related proteins, such as Cleaved Caspase 3, P53, Bcl-2 and Bax. ResultsCompared with sham surgery group, the rats in the model group showed significant degenerative changes in the intervertebral disc. The levels of aggrecan, Bcl-2, and YAP proteins in the nucleus pulposus decreased, while the levels of p-MST1/2, p-YAP, p-TAZ, P53, Bax, Cleaved Caspase 3, MMP3 proteins, and the apoptosis rate increased (P < 0.01). Compared with the model group, the drug intervention groups showed partial recovery in intervertebral disc degeneration. The levels of aggrecan, Bcl-2, and YAP proteins increased, while the levels of p-MST1/2, p-YAP, p-TAZ, P53, Bax, Cleaved Caspase 3, MMP3 proteins, and the apoptosis rate decreased (P<0.05 or P<0.01). The high-dose ZGD group showed more significant recovery in intervertebral disc degeneration compared to the low-dose ZGD group, with a decrease in the levels of p-MST1/2, p-YAP, p-TAZ, P53, Bax, Cleaved Caspase 3, MMP3 proteins, and apoptosis rate, and an increase in the levels of aggrecan, Bcl-2, and YAP proteins (P<0.05 or P<0.01). Compared with the high-dose ZGD group, the high-dose ZGD + inhibitor group showed a reduced recovery in intervertebral disc degeneration, with an increase in the levels of p-MST1/2, p-YAP, p-TAZ, P53, Bax, Cleaved Caspase 3, MMP3 proteins, and apoptosis rate, and a decrease in the levels of aggrecan, Bcl-2, and YAP proteins (P<0.05 or P<0.01). ConclusionZGD may delay intervertebral disc degeneration by inhibiting the phosphorylation of YAP in the nucleus pulposus, maintaining the function of the Hippo-YAP/TAZ signaling pathway, and reducing apoptosis of nucleus pulposus cells.
6.Protective effect of Shenfu injection against neonatal hypoxic-ischemic brain injury by inhibiting the ferroptosis
Xiaotong Zhang ; Meng Zhang ; Gang Li ; Yang Hu ; Yajing Xun ; Hui Ding ; Donglin Shen ; Ming Wu
Acta Universitatis Medicinalis Anhui 2025;60(1):31-40
Objective :
To observe the brain tissue injury during hypoxia-ischemia, as well as the pathological changes and the expression of ferroptosis-related factors after the use of Shenfu injection(SFI), and to explore the protective effect of SFI on hypoxic-ischemic brain injury(HIBD) by inhibiting ferroptosis.
Methods :
An animal model of HIBD in SD rats was constructed and intervened with SFI. Pathologic changes in brain tissue were observed by HE staining methods. Nissen staining was used to observe neuron survival. Glutathione Peroxidase 4(GPX4) and Divalent Metal Transporter 1(DMT1) expression were detected in brain tissue by Western blot, immunohistochemistry and immunofluorescence. Reduced Glutathione(GSH), Lactate Dehydrogenase(LDH), Malondialdehyde(MDA), Superoxide Dismutase(SOD) and tissue iron content were determined with the kits. BV-2 microglial cell line(BV2) cells were culturedin vitroand divided into control group(Ctrl group), oxygen-glucose deprivation group(OGD group), iron ferroptosis-inducing group(Erastin group), iron ferroptosis-inhibiting group(Fer-1 group), Shenfu injection group(SFI group), and Erastin+Shenfu injection group(Erastin+SFI group). 2′,7′-Dichlorodihydrofluorescein diacetate(DCFH-DA) reactive oxygen species(ROS) fluorescent probe was used to detect the ROS release level; Immunofluorescence was used to observe intracellular GPX4, DMT1 expression.
Results :
Compared with the Sham group, rats in the HIBD group showed significant neuronal cell damage in brain tissue, decreased GPX4 expression(P<0.01), increased DMT1 expression(P<0.01), decreased GSH and SOD levels(P<0.01), and increased LDH, MDA and tissue iron levels(P<0.05,P<0.05,P<0.01). In contrast, after the intervention of SFI, GPX4 expression was elevated(P<0.01), DMT1 expression decreased(P<0.01), GSH and SOD levels were elevated(P<0.01), and LDH, MDA, and tissue iron levels decreased(P<0.05,P<0.05,P<0.01). The cells experiments showed that compared with the Ctrl group, the OGD group had a significantly higher ROS content and a decrease in the expression of GPX4 fluorescence intensity, and an increase in the fluorescence intensity of DMT1(P<0.01), compared with the OGD group, the ROS content was reduced in the SFI group, while the expression of GPX4 was elevated and the expression of DMT1 was reduced(P<0.01).
Conclusion
Hippocampal and cortical regions are severely damaged after HIBD in neonatal rats, and their brain tissues show decreased expression of GPX4 and increased expression of DMT1. The above suggests that ferroptosis is involved in HIBD brain injury in neonatal rats. In contrast, Shenfu injection has a protective effect on HIBD experimental animal model and BV2 cell injury model by reducing iron aggregation and ROS production.
7.The renoprotective effect of Salvia miltiorrhiza injection on HBOC-CHP01 resuscitated haemorrhagic shock rats
Shasha HAO ; Xintong XIE ; Shen LI ; Honghui ZHANG ; Hong WANG ; Chengmin YANG ; Wentao ZHOU ; Jiaxin LIU
Chinese Journal of Blood Transfusion 2025;38(2):172-180
[Objective] To investigate the protective effect of Salvia miltiorrhiza injection (SMI) on the kidneys of HBOC-CHP01 resuscitated haemorrhagic shock rats. [Methods] A 50% haemorrhagic shock rat model was established, with 12 rats divided into two groups: SMI + HBOC-CHP01 group and HBOC-CHP01 group, with 6 rats in each group. The rats in the SMI+ HBOC-CHP01 group were given an equal volume of HBOC-CHP01 for resuscitation after haemorrhagic shock, and an 8 mL/kg dose of SMI. Rats in the HBOC-CHP01 group were resuscitated by administering an equilibrium blood loss volume of HBOC-CHP01 and given an 8 mL/kg dose of 0.9% NaCl solution. Blood was taken from rats at five points: before bloodletting (baseline), during haemorrhagic shock (HS), immediately after resuscitation (RS0h), 1 h after resuscitation (RS1h), and 24 h after resuscitation (RS24h). A blood gas analyser was used to detect the lactate level (Lac), glucose content (Glu), residual base (BEecf), pH, bicarbonate (HCO3-), high iron haemoglobin (MetHb). White blood cells (WBC), platelets (PLT), haemoglobin content (Hb), carboxyhaemoglobin (COHb) were detected using a quintuple classification. Blood creatinine (SCr), uric acid (UA), kidney-related indexes were detected using biochemistry instrument. Kidney tissues of the rats were taken after 24 h of resuscitation and after execution, and the inflammation of kidneys of the rats of the two groups was analyzed using HE staining. Fluorescence staining was used to detect the level of ROS in the kidneys of rats in both groups. [Results] At RS 0h, the Beecf, Glu and Lac levels of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group, and the pH level of rats in the SMI+HBOC-CHP01 group was significantly higher than that of rats in the HBOC-CHP01 group, and the Glu levels of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group at RS 1h. At RS 0h, the WBC, PLT and COHb contents of rats in the SMI+HBOC-CHP01 group were all significantly higher than those of rats in the HBOC-CHP01 group, and at RS 1h, the WBC content of rats in the SMI+HBOC-CHP01 group was significantly higher than that of rats in the HBOC-CHP01 group; at RS 1h, the UA content of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC-CHP01 group; at RS 24h, the SCr content of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC-CHP01 group; at RS 24h, the inflammation level of kidney tissues of rats in the SMI+HBOC-CHP01 group was significantly lower than that of rats in the HBOC -CHP01 group rats, and the ROS and MPO levels in the kidney tissues of rats in the SMI+HBOC-CHP01 group were significantly lower than those of rats in the HBOC-CHP01 group. [Conclusion] The combination of Salvia miltiorrhiza injection during the resuscitation of rats with severe haemorrhagic shock by HBOC-CHP01 can alleviate renal injury by reducing inflammatory response and oxidative stress.
8.Research on the Mental Model of Modern Traditional Chinese Medicine and Its Reshaping Strategies Under the Theoretical Paradigm of The Inner Canon of Yellow Emperor (《黄帝内经》)
Yaoyao HUANG ; Jie LI ; Junlong SHEN
Journal of Traditional Chinese Medicine 2025;66(7):663-668
The Inner Canon of Yellow Emperor (《黄帝内经》) constructs its theoretical system under the influence of the holistic, symbolic, and holographic mental models of traditional Chinese medicine (TCM). It lays the theoretical foundation of TCM and provides a paradigm for TCM theory construction, serving as an original academic gene and theoretical source for the inheritance and innovation of modern TCM. Based on the theoretical paradigms derived from The Inner Canon of Yellow Emperor, the holistic mental model, the symbolic mental model, and the holographic mental model in shaping nature-human medicine theory, imagery-based theory, and fractal theory, this study revealed that TCM theory relies on the cognitive framework of traditional cultural mental models. On this basis, we proposed that the modern TCM mental model should return to its roots while fostering innovation, integrating tradition with modernity by combining macroscopic with microscopic perspectives, scientific with humanistic cognition, as well as evidence-based research with digital and artificial intelligence technologies. Furthermore, with a future-oriented approach, this paper discussed on the strategies for reshaping the modern TCM mental model from three dimensions of TCM diagnosis and treatment, TCM education, and TCM research, which could not only preserve the traditional clinical thinking characteristics of TCM, but also innovate modern TCM practice.
9.Mechanism of action of bile-gut axis in the development and progression of intrahepatic cholangiocarcinoma
Xue YU ; Tianhao SHEN ; Cheng ZHOU ; Yu LIU ; Wei LI ; Tinghui JIANG ; Yongqiang ZHU ; Yan LIU
Journal of Clinical Hepatology 2025;41(3):588-593
Intrahepatic cholangiocarcinoma is a malignant tumor with an extremely poor prognosis, and its pathogenesis is complex and remains unclear. In recent years, more and more studies have focused on the role of bile-gut axis in the development and progression of intrahepatic cholangiocarcinoma. Bile-gut axis refers to the complex interaction between bile and gut microbiota, including bile salt metabolism, dynamic changes of microbiota, inflammatory response, and immune system regulation. This article elaborates on the potential mechanisms of bile-gut axis in intrahepatic cholangiocarcinoma, especially gut microbiota dysbiosis, abnormal bile salt metabolism, chronic inflammatory response, and immune system interaction, this article aims to provide new perspectives and possible therapeutic targets for future research and promote the early diagnosis and effective treatment of intrahepatic cholangiocarcinoma.
10.Analysis of Mechanism of Astragaloside Ⅳ in Regulating Ferroptosis Through SLC7A11/GPX4 Pathway Against Vascular Smooth Muscle Cell Proliferation
Guoting LI ; Changchao YANG ; Lin LIU ; Weikang LI ; Zixian ZHAO ; Quan SHEN ; Jingshan ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(10):159-167
ObjectiveTo investigate the effect of astragaloside Ⅳ(AS-Ⅳ) on the proliferation of vascular smooth muscle cells(VSMCs) induced by angiotensin Ⅱ(Ang Ⅱ) based on solute carrier family 7 member 11/glutathione peroxidase 4(SLC7A11/GPX4) pathway. MethodsPrimary rat thoracic aortic VSMCs were cultured by tissue explant method, and the cell types were identified by immunofluorescence. Cell counting kit-8(CCK-8) was used to determine the optimal concentration and time of AS-Ⅳ after Ang Ⅱ stimulation. The experiment was divided into blank group, model group, AS-Ⅳ group(40 μmol·L-1), Erastin group(0.5 μmol·L-1), Erastin+AS-Ⅳ group(0.5 μmol·L-1+40 μmol·L-1). The blank group was cultured in normal medium, the model group was cultured in medium containing Ang Ⅱ(0.1 μmol·L-1), and each administration group was cultured in medium containing Ang Ⅱ(0.1 μmol·L-1) and the corresponding doses of drug. CCK-8 and plate clone formation assay were used to detect the proliferation of cells in each group, Prussian blue staining was used to detect cell iron deposition, the content of reactive oxygen species(ROS) in cells was detected by fluorescence probe method, the content of malondialdehyde(MDA) was detected by thiobarbituric acid(TBA) method, and the protein levels of SLC7A11 and GPX4 in each group were detected by Western blot. ResultsPrimary rat thoracic aortic VSMCs were successfully cultured by tissue explant method, and immunofluorescence detection showed that positive expression of α-smooth muscle actin(α-SMA) and negative expression of vimentin in the cells, identifying them as VSMCs. The optimal concentration and time of AS-Ⅳ determined by CCK-8 were 40 μmol·L-1 and 24 h, respectively. Pharmacodynamic studies showed that compared with the blank group, the cell proliferation in the model group increased, the iron deposition in the cells increased, the contents of ROS and MDA increased, and the expression levels of SLC7A11 and GPX4 proteins decreased(P<0.05, P<0.01). Compared with the model group, the cell proliferation of the AS-Ⅳ group was inhibited, the iron deposition in the cells was decreased, the contents of ROS and MDA were decreased, and the expression levels of SLC7A11 and GPX4 proteins were increased(P<0.05, P<0.01). While in the Erastin group, the cell proliferation was increased, the iron deposition was increased, ROS and MDA contents were increased, and the expression levels of SLC7A11 and GPX4 proteins were decreased(P<0.05, P<0.01). Compared with the AS-Ⅳ group, Erastin+AS-Ⅳ group showed increased cell proliferation, increased iron deposition in cells, increased ROS and MDA contents, and decreased expression of SLC7A11 and GPX4 proteins(P<0.05). Compared with the Erastin group, the cell proliferation in Erastin+AS-Ⅳ group was inhibited, the iron deposition was decreased, the contents of ROS and MDA were decreased, and the expression levels of SLC7A11 and GPX4 proteins were increased(P<0.05, P<0.01). ConclusionAS-Ⅳ can inhibit ferroptosis by regulating the SLC7A11/GPX4 pathway, so as to weaken the proliferation of VSMCs, thus playing a role in the treatment of atherosclerosis.


Result Analysis
Print
Save
E-mail