The goal of this paper is investigating the impact and mechanism of Shenfu injection (a Traditional Chinese Medicine injection type) on prevention and therapy of paclitaxel chemotherapy in peripheral nerve harm.MethodsWistar rat dorsal root ganglion cells had been cultured in vitro and divided into teams of MOCK, PT, PT + LD, and PT + HD.
Each group was cultured at a complete serum focus of 10%, together with 10% clean serum in the MOCK group, 0.73 (IC30) μmol/L paclitaxel + 10% clean serum in the PT group, and 10% and 5% drug-containing serum and equal quantity of paclitaxel had been added into the high- and low-dosage teams, respectively.
After culturing for 24 hours, the following assessments had been carried out: (1) cell proliferation detected through the use of CCK-8 and a microplate reader; (2) axon size detected by mobile immunostaining and detection evaluation on antibody β-tubulin III; and (3) adjustments in mitochondrial membrane potential by analyzing immunofluorescence staining with JC-1 probe.Results(1) Cell proliferation: OD values of the MOCK group and PT group had been 0.43 ± 0.02 and 0.25 ± 0.03, respectively (P < 0.05), whereas OD values of teams PT + LD and PT + HD had been 0.41 ± 0.05 and 0.46 ± 0.03, respectively, greater than group PT (P < 0.05), whereas OD values of teams PT + LD and PT + HD had been 0.41 ± 0.05 and 0.46 ± 0.03, respectively, greater than group PT (μmol/L paclitaxel + 10% clean serum in the PT group, and 10% and 5% drug-containing serum and equal quantity of paclitaxel had been added into the high- and low-dosage teams, respectively.
After culturing for 24 hours, the following assessments had been carried out: (1) cell proliferation detected through the use of CCK-8 and a microplate reader; (2) axon size detected by mobile immunostaining and detection evaluation on antibody μmol/L paclitaxel + 10% clean serum in the PT group, and 10% and 5% drug-containing serum and equal quantity of paclitaxel had been added into the high- and low-dosage teams, respectively.
After culturing for 24 hours, the following assessments had been carried out: (1) cell proliferation detected through the use of CCK-8 and a microplate reader; (2) axon size detected by mobile immunostaining and detection evaluation on antibody P < 0.05), whereas OD values of teams PT + LD and PT + HD had been 0.41 ± 0.05 and 0.46 ± 0.03, respectively, greater than group PT (P < 0.05), whereas OD values of teams PT + LD and PT + HD had been 0.41 ± 0.05 and 0.46 ± 0.03, respectively, greater than group PT (.Shenfu injection can forestall the toxicity of DRG neurons induced by paclitaxel, and its mechanism could also be associated to the alleviation of mitochondrial dysfunction.
Description: DAPI (4',6-diamidino-2-phenylindole) is a fluorescent dye which can bind DNA strands robustly, the fluorescence being detected by fluorescence microscope. DAPI can dye both live and fixed cells as it can cross intact membrane, with higher efficiency in fixed cells. The molecular formula is C16H15N5·2HCl with 350.25 g/mol molecular weight, CAS Number 28718-90-3. DAPI could pass through the cell and nucleic membranes and bind the double-strand DNA in the nucleus, producing 20 times stronger fluorescence than itself. The efficiency detected by fluorescence microscope is very high (almost 100%), having no side effects for the live cells. The sensitivity for double stranded DNA DAPI staining is many times larger comparing to ethidium bromide (EB). DAPI staining is usually used in cell death detection, as it enters more effectively and generates stronger fluorescence in dead cells. After staining with DAPI, detect with fluorescence microscope or flow cytometry. Blue fluorescent cell would be seen under the microscope after staining. The largest excitation wavelength for DAPI is 340nm (ultraviolet), and the largest emission wavelength is 488nm (blue). When DAPI binds with double-strand DNA, the largest excitation wavelength is 360nm, while the largest emission wavelength becomes 460nm. DAPI's blue emission makes it suitable for combined assays where the fluorescence ranges of DAPI and other IHC-employed fluorescent molecules like green-fluorescent fluorescein and GFP, or red-fluorescent stains like Texas Red, are completely distinctive.
DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) for nucleic acid staining
Description: Abbkine Cell Cycle Staining Kit provides a convenient and accurate determination of the percentage of cells in each phase of the cell cycle.
Description: Abbkine Cell Cycle Staining Kit provides a convenient and accurate determination of the percentage of cells in each phase of the cell cycle.