Application of harpagoside in pharmacy
A technology of harpagoside and drug, applied in the pharmaceutical field of harpagoside, can solve the problem that the effect of PD cell model and PD overall animal model has not been reported yet.
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Embodiment 1
[0083] Example 1: Protection of harpagoside against MPTP + induced neuronal damage
[0084] Method: The culture of dopamine neurons in the midbrain was the same as before, and the treatment was as follows figure 1 A's experimental protocol.
[0085] Determination of cell viability: MTT method was used. Wash the cultured cells once with PBS, add MTT (0.5mg / ml) to each well, incubate for 4 hours, discard the supernatant, add DMSO, shake on the shaker for 15 minutes to completely dissolve, and read the absorbance at 490nm with a microplate reader. Cell viability = (without MPTP + Group OD-plus MPTP + group OD) / (without MPTP + group OD-MPTP + Group OD) × 100%, to obtain the change value of the number of viable cells.
[0086] Results: MPTP was observed in the experiment of cell viability assay + Toxic effect on primary nerve cell damage, adding different concentrations of harpagoside (10 -6 M, 10 -5 M) The percentage of surviving cells increased from 65.0% to 80.9% and 8...
Embodiment 2
[0091] Embodiment 2: Harpagoside to MPP + Protective effect of injured midbrain dopamine neurons
[0092] Immunocytochemical staining of midbrain dopamine neuron tyrosine hydroxylase (tyrosine hydroxylase, TH), the change of TH positive (+) cell number is the hallmark enzyme of PD. figure 2 A is to adopt figure 1A picture of the experimental protocol obtained. Observe under a Nikon inverted microscope, take 10 pictures for each culture well under a 100-fold magnification field of view, count the number of TH-positive neurons, and measure the longest process of each TH-positive neuron in the field of view under a 200-fold magnification , used to assess the neurite length of TH-positive neurons.
[0093] figure 2 B shows an increase in the number of midbrain TH(+) neurons as the concentration of harpagoside increased from 0.1 μM to 10 μM. 1 μM and 10 μM harpagoside make MPP + The relative number of injured midbrain TH(+) neurons increased from (0.604±0.010) to (0.739±0.0...
Embodiment 3
[0096] Embodiment 3: Harpagoside to MPP + Repair of damaged midbrain dopamine neurons (therapy)
[0097] image 3 A is to adopt figure 1 Experimental scheme of B, immunocytochemical staining to obtain pictures of dopamine neurons in the midbrain. Counting the number and neurite length of TH-positive neurons showed that 10 μM harpagoside in MPP + Added to the cell culture system 24 hours after injury, as the concentration of harpagoside increased from 0.1 μM to 100 μM, the number of midbrain TH(+) neurons ( image 3 B) and protrusion length ( image 3 C) shows an increasing trend. 10 μM and 100 μM harpagoside made MPP + The relative number of injured midbrain TH(+) neurons increased from (0.699±0.020) to (0.781±0.021) and (0.824±0.018), respectively increased by 11.7% (P+ The number and neurite length of injured midbrain TH(+) neurons were not significantly affected (both P>0.05).
[0098] The above results prove that the MPP + The damage was then treated with harpagosi...
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