Application of ophiocordyceps sinensis to preparation of medicines for preventing or treating pulmonary hypertension
A technology of pulmonary arterial hypertension and cordycepic acid, which can be used in drug combinations, active ingredients of hydroxyl compounds, respiratory system diseases, etc., can solve problems such as Cordyceps sinensis that have not yet been found, and achieve the effects of alleviating symptoms, reducing apoptosis, and promoting proliferation.
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Embodiment 1
[0025] Example 1 Interventional effect of cordycepic acid on the activity of pulmonary artery endothelial cells
[0026] 1. Experimental materials
[0027] Experimental subjects: Human pulmonary artery endothelial cells (HPAECs) were purchased from Science Cell Company. The sixth-generation HPAECs in logarithmic growth phase were used. They were inoculated after trypsin digestion. When the cells grew to about 70%, serum-free DMEM was used. Low-glucose medium was starved for 18 h, then normoxia, hypoxia, hypoxia + drug intervention respectively.
[0028] Main reagents: Cordyceps acid, fetal bovine serum, PBS (phosphate buffered solution), trypsin, DMEM low glucose medium, ECM complete medium, CCK-8 kit.
[0029] Main instruments: cell culture dish, cell incubator, 96-well plate, microscope, biological safety cabinet, alcohol lamp, Pasteur tube, 15ml / 50ml centrifuge tube, pipette, alcohol cotton ball, microplate reader.
[0030] 2. Experimental method
[0031] HPAEC cells in ...
Embodiment 2
[0036] Example 2 Detection of apoptosis by Annexin V-FITC / PI double staining method
[0037] 1. Experimental materials
[0038] Experimental subjects: Human pulmonary artery endothelial cells (HPAECs) were purchased from Science Cell Company. The sixth-generation HPAECs in logarithmic growth phase were used. They were inoculated after trypsin digestion. When the cells grew to about 70%, serum-free DMEM was used. Low-glucose medium was starved for 18 h, then normoxia, hypoxia, hypoxia + drug intervention respectively.
[0039] Main reagents: Cordyceps acid, fetal bovine serum, PBS, trypsin, DMEM low glucose medium, ECM complete medium, Annexin V kit.
[0040]Main instruments: cell culture dish, cell incubator, 96-well plate, microscope, biological safety cabinet, alcohol lamp, Pasteur tube, 15ml / 50ml centrifuge tube, pipette, alcohol cotton ball, microplate reader.
[0041] 2. Experimental method
[0042] Inoculate HPAEC cells in logarithmic growth phase with ECM complete me...
Embodiment 3
[0045] Example 3 TUNEL method detects the effect of cordycepin on cell damage
[0046] 1. Experimental materials
[0047] Experimental subjects: Human pulmonary artery endothelial cells (HPAECs) were purchased from Science Cell Company. The sixth-generation HPAECs in logarithmic growth phase were used. They were inoculated after trypsin digestion. When the cells grew to about 70%, serum-free DMEM was used. Low-glucose medium was starved for 18 h, then normoxia, hypoxia, hypoxia + drug intervention respectively.
[0048] Main reagents: Cordyceps acid, fetal bovine serum, PBS, trypsin, DMEM low glucose medium, ECM complete medium, in situ apoptosis detection kit (Roche, Switzerland).
[0049] Main instruments: cell culture dish, cell incubator, 96-well plate, microscope, biological safety cabinet, alcohol lamp, Pasteur tube, 15ml / 50ml centrifuge tube, pipette, alcohol cotton ball, microplate reader.
[0050] 2. Experimental method
[0051] Inoculate HPAEC cells in logarithmic...
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