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1.
Int J Mol Med ; 50(3)2022 09.
Artículo en Inglés | MEDLINE | ID: mdl-35856410

RESUMEN

The investigation of effective therapeutic drugs for pulmonary hypertension (PH) is critical. KIR2.1 plays crucial roles in regulating cell proliferation and migration, and vascular remodeling. However, researchers have not yet clearly determined whether KIR2.1 participates in the proliferation and migration of pulmonary artery smooth muscle cells (PASMCs) and its role in pulmonary vascular remodeling (PVR) also remains elusive. The present study aimed to examine whether KIR2.1 alters PASMC proliferation and migration, and participates in PVR, as well as to explore its mechanisms of action. For the in vivo experiment, a PH model was established by intraperitoneally injecting Sprague­Dawley rats monocrotaline (MCT). Hematoxylin and eosin staining revealed evidence of PVR in the rats with PH. Immunofluorescence staining and western blot analysis revealed increased levels of the KIR2.1, osteopontin (OPN) and proliferating cell nuclear antigen (PCNA) proteins in pulmonary blood vessels and lung tissues following exposure to MCT, and the TGF­ß1/SMAD2/3 signaling pathway was activated. For the in vitro experiments, the KIR2.1 inhibitor, ML133, or the TGF­ß1/SMAD2/3 signaling pathway blocker, SB431542, were used to pre­treat human PASMCs (HPASMCs) for 24 h, and the cells were then treated with platelet­derived growth factor (PDGF)­BB for 24 h. Scratch and Transwell assays revealed that PDGF­BB promoted cell proliferation and migration. Immunofluorescence staining and western blot analysis demonstrated that PDGF­BB upregulated OPN and PCNA expression, and activated the TGF­ß1/SMAD2/3 signaling pathway. ML133 reversed the proliferation and migration induced by PDGF­BB, inhibited the expression of OPN and PCNA, inhibited the TGF­ß1/SMAD2/3 signaling pathway, and reduced the proliferation and migration of HPASMCs. SB431542 pre­treatment also reduced cell proliferation and migration; however, it did not affect KIR2.1 expression. On the whole, the results of the present study demonstrate that KIR2.1 regulates the TGF­ß1/SMAD2/3 signaling pathway and the expression of OPN and PCNA proteins, thereby regulating the proliferation and migration of PASMCs and participating in PVR.


Asunto(s)
Hipertensión Pulmonar , Arteria Pulmonar , Animales , Becaplermina/metabolismo , Becaplermina/farmacología , Proliferación Celular , Humanos , Hipertensión Pulmonar/metabolismo , Monocrotalina , Miocitos del Músculo Liso/metabolismo , Antígeno Nuclear de Célula en Proliferación/metabolismo , Arteria Pulmonar/metabolismo , Ratas , Ratas Sprague-Dawley , Factor de Crecimiento Transformador beta1/metabolismo , Remodelación Vascular
2.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 38(5): 543-548, 2022 Sep.
Artículo en Chino | MEDLINE | ID: mdl-37088767

RESUMEN

OBJECTIVE: To investigate whether probenecid (PROB) could improve the proliferation and migration ability of rats' pulmonary artery smooth muscle cells induced by platelet-derived growth factor-BB (PDGF-BB). METHODS: Primary pulmonary artery smooth muscle cells (PASMCs) of SD rats were cultured in vitro, and were randomly divided into control group (CON group), PDGF-BB group (10 ng/ml PDGF-BB treatment for 24 h) and PDGF-BB+PROB group (10 ng/ml PDGF-BB and 200 µmol/L PROB treatment for 24 h, PROB is a specific blocker of pannexin-1). CCK-8 method was used to select the suitable intervention concentrations of PROB and PDGF-BB, and to detect the proliferation of PASMCs in each group. The migration ability of PASMCs was detected by TranswellTM assay and cell scratch test. Immunofluorescence cytochemistry and Western blot were used to detect the protein expressions and distribution of osteopontin (OPN) and proliferating cell nuclear antigen (PCNA) in PASMCs. RESULTS: Compared with CON group, the migration and proliferation ability of PASMCs in PDGF-BB group were enhanced (P<0.05). After treated with PROB, the migration and proliferation ability of PASMCs in PDGF-BB+PROB group were decreased significantly (P<0.05). Compared with CON group, the expression and protein levels of OPN and PCNA in PDGF-BB group were increased significantly (P<0.05), while the expression and protein levels of OPN and PCNA in PDGF-BB+PROB were decreased significantly (P<0.05). CONCLUSION: Probenecid inhibits the migration and proliferation of PDGF-BB-induced PASMCs by blocking Pannexin-1.


Asunto(s)
Probenecid , Arteria Pulmonar , Ratas , Animales , Becaplermina/metabolismo , Becaplermina/farmacología , Antígeno Nuclear de Célula en Proliferación/metabolismo , Probenecid/farmacología , Probenecid/metabolismo , Proteínas Proto-Oncogénicas c-sis/farmacología , Proteínas Proto-Oncogénicas c-sis/metabolismo , Proliferación Celular , Ratas Sprague-Dawley , Miocitos del Músculo Liso , Células Cultivadas
3.
Pulm Pharmacol Ther ; 70: 102072, 2021 10.
Artículo en Inglés | MEDLINE | ID: mdl-34428599

RESUMEN

BACKGROUND: Recent studies have shown that endothelin-1 and angiotensin II (AngII) can increase gap junctional intercellular communication (GJIC) by activating Mitogen-activated protein kinases (MAPKs) pathway. However, not only the precise interaction of AngII with Connexin43(Cx43) and the associated functions remain unclear, but also the regulatory role of Cx43 on the AngII-mediated promotion proliferation and migration of VSMCs is poorly understood. MATERIAL AND METHODS: Our research applicated pressure myography measurements, immunofluorescence and Western blot analyses to investigate the changes in physiological indicators in spontaneously hypertensive rats (SHRs) and AngII-stimulated proliferation and migration of A7r5 SMCs(Rat vascular smooth muscle cells). The aim was to elucidate the role of CX43 in hypertension induced by AngII. RESULTS: Chronic ramipril (angiotensin converting enzyme inhibitor) management for SHRs significantly attenuated blood pressure and blood vessel wall thickness, also reduced contraction rate in the cerebral artery. The cerebral artery contraction rates, mRNA and protein expression of Cx43, osteopontin (OPN) and proliferating cell nuclear antigen (PCNA) protein expression in the SHR + ramipril and SHR + ramipril + carbenoxolone (CBX, Cx43 specific blocker) groups were significantly lower than those in the SHR group. Cx43 protein expression and Ser368 phosphorylated Cx43 protein levels increased significantly in AngII-stimulated A7r5 cells. However, the levels of phosphorylated Cx43 decreased after pre-treatment with candesartan (AT1 receptor blocker), GF109203X (protein kinase C (PKC) blocker) and U0126 (mitogen-activated protein kinases/extracellular signal-regulated kinase1/2(MEK/ERK1/2)-specific blocker) in AngII-stimulated A7r5 cells. Cx43 was widely distributed in the cell membrane, nucleus, and cytoplasm of the SMCs. Furthermore, pre-treatment of the AngII- stimulated A7r5 cells with Gap26 (Cx43 blocker) significantly inhibited cell migration and decreased the expression levels of MEK1/2, ERK1/2, P-MEK1/2, and P-ERK1/2. CONCLUSION: Our research confirms that Cx43 plays an important role in the regulation of proliferation and migration of VSMCs via MEK/ERK and PKC signal pathway in AngII-dependent hypertension.


Asunto(s)
Angiotensina II , Conexina 43/fisiología , Hipertensión , Miocitos del Músculo Liso/citología , Angiotensina II/farmacología , Animales , Proliferación Celular , Músculo Liso Vascular , Ratas
4.
Front Cell Neurosci ; 15: 665596, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34113239

RESUMEN

Background Our aim was to investigate the effects of the protein expression and the function of sodium, potassium, and chloride co-transporter (NKCC1) in the dorsal root ganglion (DRG) after activation of transient receptor potential vanilloid 1 receptor (TRPV1) in capsaicin-induced acute inflammatory pain and the possible mechanism of action. Methods Male Sprague-Dawley rats were randomly divided into control, capsaicin, and inhibitor groups. The expression and distribution of TRPV1 and NKCC1 in rat DRG were observed by immunofluorescence. Thermal radiation and acetone test were used to detect the pain threshold of heat and cold noxious stimulation in each group. The expressions of NKCC1 mRNA, NKCC1 protein, and p-NKCC1 in the DRG were detected by PCR and western blotting (WB). Patch clamp and chloride fluorescent probe were used to observe the changes of GABA activation current and intracellular chloride concentration. After intrathecal injection of protein kinase C (PKC) inhibitor (GF109203X) or MEK/extracellular signal-regulated kinase (ERK) inhibitor (U0126), the behavioral changes and the expression of NKCC1 and p-ERK protein in L4 - 6 DRG were observed. Result: TRPV1 and NKCC1 were co-expressed in the DRG. Compared with the control group, the immunofluorescence intensity of NKCC1 and p-NKCC1 in the capsaicin group was significantly higher, and the expression of NKCC1 in the nuclear membrane was significantly higher than that in the control group. The expression of NKCC1 mRNA and protein of NKCC1 and p-NKCC1 in the capsaicin group were higher than those in the control group. After capsaicin injection, GF109203X inhibited the protein expression of NKCC1 and p-ERK, while U0126 inhibited the protein expression of NKCC1. In the capsaicin group, paw withdrawal thermal latency (WTL) was decreased, while cold withdrawal latency (CWL) was prolonged. Bumetanide, GF109203X, or U0126 could reverse the effect. GABA activation current significantly increased in the DRG cells of the capsaicin group, which could be reversed by bumetanide. The concentration of chloride in the DRG cells of the capsaicin group increased, but decreased after bumetanide, GF109203X, and U0126 were administered. Conclusion Activation of TRPV1 by exogenous agonists can increase the expression and function of NKCC1 protein in DRG, which is mediated by activation of PKC/p-ERK signaling pathway. These results suggest that DRG NKCC1 may participate in the inflammatory pain induced by TRPV1.

5.
Zhongguo Zhong Yao Za Zhi ; 44(14): 2926-2931, 2019 Jul.
Artículo en Chino | MEDLINE | ID: mdl-31602834

RESUMEN

Diemailing~® Kudiezi Injection( DKI) is widely used in the treatment of cerebral infarction,coronary heart disease and angina pectoris. Long-term clinical application and related research evidence showed that DKI has a good effect in improving the clinical symptoms of cardiovascular and cerebrovascular diseases. However,this injection has not been included in any clinical practice guideline. It has been found that the use of DKI is in wrong way in clinical practice in recent years. Therefore,clinical experts from the field of cardiovascular and cerebrovascular diseases nationwide are invited to compile this expert consensus in order to guide clinicians.GRADE system is used to grade the quality of evidence according to different outcomes according to degrading factors. Then it forms the recommendation or consensus suggestion through the nominal group method. The formation of expert consensus mainly considers six factors: quality of evidence,economy,efficacy,adverse reactions,patient acceptability and others. Based on these six aspects,if the evidence is sufficient,a " recommendation" supported by evidence is formed,and GRADE grid voting rule is adopted. If the evidence is insufficient,a " consensus suggestions" will be formed,using the majority voting rule. In this consensus,the clinical indications,efficacy,safety evidences and related preliminary data of DKI were systematically and comprehensively summarized in a concise and clear format,which could provide valuable reference for the clinical use of DKI. This consensus has been approved by China association of Chinese medicine which is numbered GS/CACM 202-2019.


Asunto(s)
Angina de Pecho/tratamiento farmacológico , Infarto Cerebral/tratamiento farmacológico , Enfermedad Coronaria/tratamiento farmacológico , Medicamentos Herbarios Chinos/uso terapéutico , China , Consenso , Humanos , Inyecciones , Medicina Tradicional China
6.
Chin J Integr Med ; 16(6): 493-7, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21110173

RESUMEN

OBJECTIVE: To analyze the effectiveness of Chinese medicine and integrated Chinese and Western medicine for influenza A (H1N1) in the fever clinics and its relevant expenditure. METHODS: A prospective survey on the clinical epidemic observation and follow-up was conducted from July 2009 to October 2009 with a self-developed questionnaire whose contents including the clinical data of the confirmed 149 H1N1 cases and their relevant therapeutic expenditure. The patients were assigned to the Chinese medicine group (22 cases treated by Chinese medicine alone) and integrative medicine group (124 cases treated by both Chinese medicine and Western medicine). The data were processed with descriptive analysis, t test and χ (2), and sum-rank test. RESULTS: The proportion of clinical recovery of Chinese medicine group (81.8%) was higher than that of integrative medicine group (54.8%) with statistical significance (P=0.02). The average fever durations in both groups were 3.5 to 4 days, showing no significant difference (P=0.86). In the comparisons of average cost of Chinese herbs, drugs, therapies, and total cost, those of the Chinese medicine group were lower than those in the integrative group (P=0.01, P=0.00, P=0.00, P=0.00). CONCLUSIONS: The H1N1 patients in the fever clinic who received Chinese medicine treatment had a higher clinical recovery proportion than those who received integrated Chinese and Western medicine treatment with lower medical cost. However, due to small sample size of the Chinese medicine group in the study, the conclusion needs further confirmation by studies with large sample size.


Asunto(s)
Fiebre/economía , Gastos en Salud , Subtipo H1N1 del Virus de la Influenza A/fisiología , Gripe Humana/economía , Gripe Humana/terapia , Medicina Integrativa/economía , Medicina Tradicional China/economía , Adulto , Costos y Análisis de Costo , Femenino , Fiebre/terapia , Fiebre/virología , Hospitales , Humanos , Gripe Humana/virología , Masculino , Factores de Tiempo , Resultado del Tratamiento
7.
Zhongguo Zhong Xi Yi Jie He Za Zhi ; 30(4): 343-7, 2010 Apr.
Artículo en Chino | MEDLINE | ID: mdl-20669665

RESUMEN

Based on the principle of management, evidence-based medicine, operational research and health economics, this essay addressed the theoretical basis of clinical pathway and its application of evidence-based Chinese medicine to practice. It could be taken as references for different health care institutions and organizations for development of clinical pathway.


Asunto(s)
Vías Clínicas , Medicina Basada en la Evidencia , Medicina Tradicional China , Medicina Basada en la Evidencia/organización & administración , Humanos
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