The Protection Effects Of Electroacupuncture At Shuigou Acupoint On Neurovascular Unit In Ischemic Cerebral Stroke

Objective:The aim of this study is to observe the effects of elecroacupuncture at Shuigou acupoint on cerebral infarction volume,neurological deficit scores and brain organelle in ischemic cerebral stroke in different time windows.Detect the microvessel density,blood-brain barrier function and the expression of lnc RNAs and m RNAs to study the protective effect of electroacupuncture at shuigou acupoint on rat with ischemic cerebral stroke and discuss the possible mechanism of acupuncture.Method:Sprague-Dawley male rats were randomly divided into 4 groups(control group,sham operation group,model group and EA group),and the model of MCAO was made with Zea-Longa method.30 minutes after successful modeling,rats in EA group were acupunctured at Shuigou.Longa evaluation method was used to record NDS and TTC staining was applied to calculate the cerebral infarction volume.Immunohistochemistry was used to detect the expression of microvessel density marker CD34 and BBB permeability marker AQP4 in the ischemic penumbra.The ischemic brain tissues were scanned by transmission electron microscopy to observe the apoptosis degree of brain cells in each group.High-throughput techniques and q RT-PCR were used to screen and validate differentially expressed lnc RNAs and m RNAs,respectively.Result:1.The results of TTC staining showed that the cerebral infarction volume of model group was significantly higher than that of control group or sham operation group(P <0.01).The cerebral infarction volume of EA group was significantly lower than that of model group(P < 0.01).The neurological deficit scores results showed that NDS of rats in control group and sham operation group were zero,and the NDS in EA group was significantly lower than that in model group after EA therapy(P < 0.05).2.The results of immunohistochemistry showed that the content of MVD marker CD34 in model group was significantly lower than that in sham operation group or EA group(P < 0.05).The content of BBB function marker AQP4 in model group was significantly higher than that in sham operation group or EA group(P < 0.05);Electron microscopy showed that there was no apoptosis in brain cells of sham operation group or control group,but the apoptosis of the ischemic cells in model group was severe.Slight apoptosis was observed in ischemic brain cells of EA group.3.High-throughput results showed that a total of 675 lnc RNAs and 1795 m RNAs were differentially expressed among three groups.The q RT-PCR results showed that the relative expression quantities of lnc RNA LOC100909777 and m RNA Havcr2,Arl11,and Arhgdib were significantly higher in model group when compared with sham operation group(P < 0.05).However,there was no statistical difference of the relative expression quantities of these RNAs between EA group and model group(P > 0.05).Conclusion:1.30 minutes after ICS,EA at Shuigou can effectively reduce the cerebral infarction volume of ischemic cerebral stroke rats and improve the neurological function of ischemic cerebral stroke rats.2.EA at Shuigou acupoint could effectively augment the content of CD34 in ischemic penumbra,which means the increasing number of microvessels in ischemic brain tissues.On the contrary,EA at Shuigou acupoint could reduce the content of AQP4 in ischemic penumbra to affect BBB function.EA could also protect ICS rat brain nerve cells in ischemic penumbra and prevent organelle damage.3.Eventually,lnc RNA LOC100909777 and m RNA Havcr2,Arl11,Arhgdib could be used as potential markers and therapeutic targets for ischemic cerebral stroke,but lnc RNAs and m RNAs associated with acupuncture therapy need to be further studied.