Polymer nano-carrier for folic acid specificity targeting of active hepatic stellate cell and medicinal application thereof
A technology for targeting hepatic stellate cells and folic acid, which is applied to a polymer nanocarrier targeting activated hepatic stellate cells and its preparation and application fields, and can solve the coupling that has not yet specifically targeted hepatic stellate cells. Folic acid ligand polymer nanocarriers and other issues
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Embodiment 1
[0027] Isolation and Culture of Primary Rat Hepatic Stellate Cells (HSC)
[0028] Rat liver was digested by in situ perfusion method, and the primary rat hepatic stellate cells obtained by digestion were placed in high-glucose DMEM medium (containing 15%-20% FBS, 1% double antibody P / S) at 37°C , 5% CO 2 cultured in a cell incubator.
Embodiment 2
[0030] Culture of Activated Rat Hepatic Stellate Cells (HSC-T6)
[0031] After recovery and passage of HSC-T6 cells, add high-glucose DMEM medium (containing 10% FBS, 1% double antibody P / S) and place at 37°C, 5% CO 2 cultured in a cell incubator.
Embodiment 3
[0033] Analysis of mRNA expression changes of primary rat hepatic stellate cells (HSC) from resting state to activated state αSMA, Collagen1a1, FR-α
[0034] Rat liver was digested by in situ perfusion method, and the primary rat hepatic stellate cells obtained from the digestion were cultured in a 24-well plate for 7 days, and then the culture medium of the 24-well plate was aspirated, washed twice with PBS, and the PBS was discarded . Axygen kit was used to extract primary rat hepatic stellate cell RNA, reverse transcription kit and QPCR instrument were used to reverse transcribe the RNA into cDNA and dilute it, and then perform PCR to analyze the mRNA expression of αSMA, Collagen1a1 and FR-α Variety. The changes in mRNA expression are shown in Figure 1 (a, b, c).
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