Effect Of Head Acupoint Pressing On TLR4/NF-κB Signaling Pathway In Rats With Cerebral Ischemia Injury

Objective: To investigate the effects of head acupoint pressing on the expression levels of TLR4、NF-κBp65 and the downstream inflammatory factors TNF-α and IL-10 in brain tissue by preparing a middle cerebral artery occlusion(MCAO)model rat,and to investigate the mechanism of head acupoint pressing to reduce the inflammatory response in rats with cerebral ischemic injury.Methods: 36 healthy male SD rats weighing 260-300 g were divided into sham operation group(n=12)and operation group(n=24)according to the random number table method.The right middle cerebral artery occlusion model was established by the thread embolization method in the operation group of rats,after which the rats with successful modeling were randomly divided into the model group(n=12)and the head acupoint pressing group(n=12).Head acupoint pressing group were given to spoon needles on Baihui and Shenting of rats 2h after modeling,1time/d,10min/time,for 7 days.The sham operation group and model group were only captured in the same way and did not intervene.The m NSS score was used to assess the neurological function of rats;TTC staining was used to detect the area of cerebral infarction in rats;HE staining was used to observe the morphological changes of brain tissue in rats;RT-q PCR and Western Blot techniques were used to detect the expression levels of TLR4 and NF-κBp65 in rat cerebral cortex;ELISA was used to detect the expression levels of TNF-α and IL-10 in rat cerebral cortex.Results:1.m NSS sore: The rats in the sham operation group were free of neurological deficits,while the rats in the model group and the head acupoint pressing group showed symptoms of neurological deficits before treatment,with no significant difference between them(P>0.05).After 7 days of intervention,compared with the model group,the neurological deficit symptoms of the rats in the head acupoint pressing group were significantly reduced(P<0.05).2.TTC staining: The rats in the sham operation group had no ischemic infarction,and the rats in the model group and the head acupoint pressing group both showed white infarct foci,but the infarct area in the head acupoint pressing group was significantly smaller than that in the model group(P<0.05).3.HE staining: The neuronal cells in the sham operation group were structurally intact;the neuronal cells in the model group were loosely structured and disorganized,and a large number of cells were seen to be deformed,thickly stained,edematous and ruptured;cells with abnormal morphology were also seen in the head acupoint pressing group,but the number was reduced compared with that of the model group.4.RT-q PCR: Compared with the sham operation group,the m RNA expression levels of TLR4 and NF-κBp65 in the cerebral cortex on the infarct side of the rats in the model group were significantly higher(P<0.05);compared with the model group,the m RNA expression levels of TLR4 and NF-κBp65 in the cerebral cortex on the infarct side of the rats in the head acupoint pressing group were significantly lower(P<0.05).5.Western Blot: Compared with the sham operation group,the protein expression levels of TLR4 and NF-κBp65 in the cerebral cortex of the infarcted side of the rats in the model group were significantly higher(P<0.05);compared with the model group,the protein expression levels of TLR4 and NF-κBp65 in the cerebral cortex of the infarcted side of the rats in the head acupoint pressing group were significantly lower(P<0.05).6.ELISA: Compared with the sham operation group,the levels of TNF-α and IL-10 in the cerebral cortex of the infarcted side of the rats in the model group were significantly up-regulated(P<0.05),while the levels of TNF-α and IL-10 in the cerebral cortex of the infarcted side of the rats in the head acupoint pressing group were significantly down-regulated and IL-10 was significantly up-regulated compared with the model group(P<0.05).Conclusion:1.Head acupoint pressing can reduce the symptoms of neurological deficits,decrease the area of cerebral infarction and improve the morphology of the affected brain tissue in MCAO rats,indicating that head acupoint point pressing has neuroprotective effects in cerebral ischemia.2.The neuroprotective effect of head acupoint pressing on cerebral ischemia may be achieved by inhibiting TLR4 and NF-κBp65,key signaling molecules on the inflammatory signaling pathway TLR4/NF-κB,as well as down-regulating the downstream pro-inflammatory factor TNF-α and up-regulating the level of the anti-inflammatory factor IL-10.