Application of butylphthalide and derivatives thereof in preparation of medicines for preventing and treating ALS
A technology of racemic butylphthalide and L-butylphthalide, which is applied in the field of medicine to achieve the effect of delaying the onset time, prolonging the survival period, and reducing degenerative diseases
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Embodiment 1
[0059] Example 1: Identification of ALS transgenic mice
[0060] The ALS transgenic mouse that is widely used in clinical and basic research is SOD1-G93A mouse, which carries the SOD1 gene that expresses the missense mutation of human G93A (the 93rd glycine in the amino acid chain is replaced by alanine), which can simulate the chronic progression of ALS The course of the disease and some pathological and biochemical manifestations. The main characteristics of SOD1-G93A mice: about 3-4 months of hindlimb weakness, hindlimb tremor and abduction weakness when the tail is suspended, after 2 weeks, one or two limbs of the mouse are paralyzed, and the hair gradually appears rough and thin, 5 Months later, he was unable to exercise and finally died of respiratory failure.
[0061] The ALS transgenic mouse of the present invention (B6SJL-TgN[SOD1-G93A]1Gur, number 002726) was purchased from Jackson Laboratory (Barharbor, ME, U.S.A).
[0062] 1.1 Identification of offspring mice
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Embodiment 2
[0071] Example 2: Observing the effects of racemic butylphthalide on the onset time, survival period and motor function of ALS mice
[0072] The mental state, disease progression and survival of all SOD1-G93A transgenic mice were observed every day. Judgment of the onset: Observation of limb tremor and / or limb weakness for 2 consecutive days was judged as the onset of death. Time of death: the rat was placed in the supine position, and the death was judged if the rat could not turn over to the prone position within 30 seconds.
[0073] Rotarod Test: The rotarod test assesses motor coordination, strength, and balance. The experimental mice were placed on a rotarod instrument with a diameter of 3.5cm, and the speed was adjusted to 16rpm. The longest latency period for falling was recorded, with 180 seconds as the cut-off value. If it exceeded 180 seconds, it was recorded as 180 seconds, and if it was less than 180 seconds, it was recorded as the actual time. The evaluation of t...
Embodiment 3
[0078] Example 3: Electrophysiological detection of ALS transgenic mice
[0079] Spontaneous potential: the experimental rat was anesthetized with 10% chloral hydrate, and the concentric needle electrode was inserted into the gastrocnemius muscle belly of the hind limb, and observed whether there was a fibrillation potential and (or) a positive sharp wave.
[0080] Recording of Compound Muscle Action Potential (CMAP): Rats were anesthetized with 10% chloral hydrate, the lumbar paraspinal sciatic nerve was stimulated, and the recording electrode was inserted into the gastrocnemius muscle belly of the hind limb. Give super strong stimulation, get CMAP and record, measure peak-to-peak value to represent CMAP amplitude, and record distal motor latency (DML).
[0081] Motor unit number (MUNE) evaluation: Referring to the principle of "statistical methods and techniques", the experimental mice were anesthetized with 10% chloral hydrate, the lumbar paraspinal sciatic nerve was stimul...
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