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OBJECTIVE: To discuss the effect of feiliuping ointment on proliferation, angiogenesis and PI3K/AKT pathway of rat bone marrow-derived endothelial progenitor cells (EPCs) in A549 lung tumor cells supernatant. METHODS: Bone marrow mononuclear cells were isolated from Sprague-Dawley rats by density gradient centrifugation methods and cultured in EGM-2 MV medium. And fluorescent immunocytochemistry was used to detect CD133 and vascular endothelial growth factor receptor 2 (VEGFR2) expression. EPCs were treated with 0.9% sodium chloride solution (blank control) or serum of rats treated with high dose and low dose Feiliuping ointment for 48 h to explore the concentration-effect, or with serum of rats treated with high dose feiliuping ointment for 0, 12, 24, 48 h to explore the time-effect. MTT assay and Matrigel method were used to detect the proliferation and angiogenesis of EPCs. The NF-κB, p-Akt, PI3K p85 proteins of EPCs were detected by Western blot. RESULTS: After induced culture for 7 d, the isolated bone marrow mononuclear cells exhibited round or short spindle-shaped appearance, were Dil-Ac-LDL and FITC-UEA-1 positive, and expressed CD133+VEGFR2+. After being treated with high dose feiliuping ointment serum for 48 h, OD490 nm value was lower than that in the control group or low dose feiliuping ointment group (P<0.05). The inhibition effect of rat serum after Feiliuping ointment administration on EPCs was dose-dependent and time-dependent. Matrigel tube formation assays showed that the counts of tube formation of EPCs in high dose Feiliuping ointment group was lower than that in control group or low dose of feiliuping ointment group (P<0.05). And the levels of NF-κB, p-Akt, PI3K p85 proteins in high dose of feiliuping ointment group were lower than those in control group(P<0.01). CONCLUSION:S Treatment with administration can Feiliuping ointment serum would inhibit the proliferation and tube formation of bone marrow-derived EPCs in A549 supernatant via the down-regulation of PI3K/AKT signaling pathway.
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Objective: To study the inhibitory effect of Pien Tze Huang on proliferation, migration, and invasion of lung cancer cells and explore its mechanism. Methods: Human lung cancer cell A549 was used as the experimental object in vitro, and the cells were treated with different concentrations of Pien Tze Huang. MTT was used to detect cell proliferation, and the changes of cell invasion and migration were measured by Transwell assay. Western blot was used to detect the phosphorylation of invasive migration proteins MMP-9, MMP-2 and EMT proteins E-cadherin, vimentin and Akt signaling pathway proteins. Lung cancer cells were treated with Akt signaling pathway activator and tablets to detect the effect of activating Akt signaling pathway on the proliferation, invasion, and migration of lung cancer cells. Results: Pien Tze Huang (100 μg/mL tablets) had no effect on the proliferation of lung cancer cells, the proliferative ability of lung cancer cells could be significantly reduced by 200 μg/mL Pien Tze Huang. The invasion and migration ability of lung cancer cells treated with 100 and 200 μg/mL tablets were decreased, the protein expressions of MMP-2 and MMP-9 in cells were decreased, the level of E-cadherin protein in cells was increased, the level of vimentin protein was decreased, at the same time, the phosphorylation level of Akt was decreased. Akt signaling pathway activator treatment could significantly reduce the inhibitory effect of Pien Tze Huang on proliferation, invasion and migration of lung cancer cells (P < 0.05). Conclusion: Pien Tze Huang decreased the proliferation, migration and invasion of lung cancer cells, the mechanism is related to the inhibition of the activation of Akt signaling pathway.
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OBJECTIVE: To discuss the effect of Qingfei-yangyin-huoxue recipe on radiation-induced pulmonary injury by regulating the lncRNA NANCI-NKX2.1 pathway. METHODS: The C57BL/6 mice were randomly divided into normal control group (A group, n=16), drug only group (B group, n=16), model group (C group, n=16) and drug model group (D group, n=16). After exposure to Qingfei-yangyin-huoxue recipe for 10 w, the pathological change of lung tissue was examined by H&E and Masson staining. The expressions of lncRNA NANCI and NKX2.1 mRNA in lung tissues were detected by qRT-PCR. And the NKX2.1 protein expressions were detected by Western blot. RESULTS: The mean animal weight in D groups was less than A and B group, but more than C group after treatment of 7 d(P<0.05). There were marked interstitial edema and inflammatory cells, fibrocytes accumulation in C group but not in A and B groups by H&E and Masson stain. The alveolitis and fibrosis changes in D group were better than C group. And the mean radiation-induced pulmonary injury score in D group was (3.875±1.746), which was less than C group, but more than A and B groups (P<0.05). The expression of lncRNA NANCI and NKX2.1in D group was higher than C group, but lower than A and B groups (P<0.05). Besides, the radiation-induced pulmonary injury score was negative related with lncRNA NANCI and NKX2.1 (r=-0.510, -0.786, P<0.05). CONCLUSION: There are significant evidences that Qingfei-yangyin-huoxue recipe could protect radiation-induced pulmonary injury by up-regulation lncRNA NANCI-NKX2.1 pathway.
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OBJECTIVE@#To study the effects of Transient receptor potential cation channel, subfamily V, member 1 (TRPV1) combined with lidocaine on status and apoptosis of U87-MG glioma cell line, and explore whether local anesthetic produces neurotoxicity by TRPV1.@*METHODS@#U87-MG cells were divided into control group, gene silencing group, empty vector group and TRPV gene up-regulation group. For cells in each group, flow cytometry was employed to detect the intracellular calcium ion concentration and mitochondrial membrane potential at different time point from cellular perspective. Cell apoptosis of U87-MG was assayed by flow cytometry and MTT from a holistic perspective.@*RESULTS@#Calcium ion concentration increased along with time. The concentration in TRPV1 gene up-regulation group was significantly higher than those in other groups at each time point (P < 0.05). After adding lidocaine, mitochondrial membrane potential in U87-MG significantly increased (P < 0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P < 0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P < 0.05). Flow cytometry result and MTT result both showed that cell apoptosis in each group significantly increased after lidocaine was added (P < 0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P < 0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P < 0.05). Moreover, apoptosis was more severe along with the increasing concentration of lidocaine (P < 0.05).@*CONCLUSIONS@#In this study, it was proved that lidocaine could dose-dependently induce the increase of intracellular calcium ion concentration, mitochondrial membrane potential and apoptosis in U87-MG glioma cell line. The up-regulation of TRPV1 enhanced cytotoxicity of lidocaine, which revealed the correlations between them. Lidocaine might have increased intracellular calcium ion concentration by activating TRPV1 gene and induced apoptosis of U87-GM glioma cell line by up-regulating mitochondrial membrane potential.
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Objective: To study the effects of Transient receptor potential cation channel, subfamily V, member 1 (TRPV1) combined with lidocaine on status and apoptosis of U87-MG glioma cell line, and explore whether local anesthetic produces neurotoxicity by TRPV1. Methods: U87-MG cells were divided into control group, gene silencing group, empty vector group and TRPV gene up-regulation group. For cells in each group, flow cytometry was employed to detect the intracellular calcium ion concentration and mitochondrial membrane potential at different time point from cellular perspective. Cell apoptosis of U87-MG was assayed by flow cytometry and MTT from a holistic perspective. Results: Calcium ion concentration increased along with time. The concentration in TRPV1 gene up-regulation group was significantly higher than those in other groups at each time point (P < 0.05). After adding lidocaine, mitochondrial membrane potential in U87-MG significantly increased (P < 0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P < 0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P < 0.05). Flow cytometry result and MTT result both showed that cell apoptosis in each group significantly increased after lidocaine was added (P < 0.05). This increasing trend in TRPV1 gene up-regulation group was more significant than other groups (P < 0.05), while in TRPV1 gene silencing group, the trend significantly decreased (P < 0.05). Moreover, apoptosis was more severe along with the increasing concentration of lidocaine (P < 0.05). Conclusions: In this study, it was proved that lidocaine could dose-dependently induce the increase of intracellular calcium ion concentration, mitochondrial membrane potential and apoptosis in U87-MG glioma cell line. The up-regulation of TRPV1 enhanced cytotoxicity of lidocaine, which revealed the correlations between them. Lidocaine might have increased intracellular calcium ion concentration by activating TRPV1 gene and induced apoptosis of U87-GM glioma cell line by up-regulating mitochondrial membrane potential.