| Objective:To observe the effect of tube moxibustion on the expression of related signal transduction molecules in PI3K/Akt and PLC/PKC pathway in rabbits with facial nerve injury,so as to explore the molecular mechanism of tube moxibustion in the treatment of peripheral facial paralysis.Methods:The model of facial nerve crush injury in rabbits was established.The experimen t was divided into two stages with 32 rabbits each experiment.Except for sampling and testing,the other operation were exactly same.Eight rabbits were selected from32 rabbits by random number table method and included in sham operation group.The remaining rabbits were randomly divided into model group,tube moxibustion group and thermotherapy group according to their body weight.Intervention treatm ent began after 5 days of successful modeling.Moxa velvet and charcoal were added to the tube moxibustion apparatus in the tube moxibustion group and thermotherapy group respectively,while no intervention treatment was given in the sham operation group and the model group.The samples were taken the next day after treatment,and the first batch of perfusion samples were used to detect the expression of Bax,Bcl-2and GAP-43 in facial nerve and facial nucleus by immunohistochemistry.The second batch of samples were taken directly for ELISA and Western Blot to detect the expre ssion of PI3Kp85,Akt1/2/3,p-Akt1,PLC,p-PLC,DAG,PKC? and p-PKC? in rabbit facial nerve.Results:1.Effect on signal transduction molecules in PI3K/Akt pathway: Compared with the model group,the expression of PI3Kp85,Akt1/2/3 and p-Akt1 protein in facial nerve in sham operation group was lower(P<0.05 or P<0.01).PI3Kp85,Akt1/2/3and p-Akt1 protein expression in facial nerve in tube moxibustion group was higher than that in model group(P<0.01),and the protein expression of PI3 K p85,Akt1/2/3and p-Akt1 in facial nerve in hyperthermia group was higher than that in model group(P<0.05 or P<0.01).2.Effect on signal transduction molecules in PLC/PKC pathway: Compared with the model group,the expression of DAG in facial nerve was lower(P < 0.05),the expression of p-PKC? protein was higher than that in sham operation group(P<0.01),and there was no significant difference in the expression of PLC,p-PLC and PKC?protein in sham operation group(P>0.05).The expression of p-PLC,DAG,PKC?and p-PKC? protein in facial nerve in tube moxibustion group and the expression of PKC? protein in facial nerve of rabbits in hyperthermia group was higher than that in sham operation group(P<0.05 or P<0.01),and there was no significant difference in other above mentioned indexes(P>0.05).3.Effects on effector molecules related to nerve repair and regeneration: Com pared with the model group,the expression of Bax protein in facial nerve in sham operation group was lower(P<0.05),but there was no significant difference in Bax,Bcl-2,GAP-43,Bcl-2 and GAP-43 protein expression in facial nucleus(P>0.05).The expression of Bcl-2 and GAP-43 protein in facial nucleus and facial nerve washigher in the tube moxibustion group(P<0.05),but there was no significant different ce in the expression of Bax protein in the facial nucleus and facial nerve in the tube moxibustion group(P>0.05),and there was no significant difference in the express ion of the above indexes in the facial nucleus and facial nerve in the hyperthermia group(P>0.05).Conclusion:1.The protein expression of PI3Kp85,Akt1/2/3,p-Akt1 and DAG in the model group was higher than that in sham operation group,suggesting that facial nerve injury can enhance the signal transduction of PI3K/Akt and PLC/PKC pathway,so as to promote the self-repair of the body.2.The protein expression of PI3Kp85,Akt1/2/3,p-Akt1 and PKC? in the hyper thermia group was higher than that in the model group,suggesting that hyperthermia can promote the repair and regeneration of injured facial nerve by enhancing the signal transduction of PI3K/Akt and PLC/PKC pathway.3.The expressions of PI3Kp85,Akt1/2/3,p-Akt1,p-PLC,DAG,PKC?,p-PKC?and effect factors Bcl-2 and GAP-43 in the tube moxibustion group were higher than those in the model group,suggesting that tube moxibustion can promote the repair and regeneration of injured facial nerve by enhancing the signal transduction of PI3K/Akt and PLC/PKC pathway.4.The effect of tube moxibustion is more extensive and the trend is stronger than that of hyperthermia.Therefore,it can be considered that the therapeutic effect of tube moxibustion is better than that of thermotherapy. |
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