Effect Of Fructus Aurantii Immaturus On The Structure And Function Of Interstitial Cells Of Cajal In Rats With Cerebral Infarction

Objective：1、To investigate the potential mechanism of acute ischemic stroke withgastrointestinal dysfunction diseases from the cell’s point of view;2、To assess the efficacy of drugs for regulating Qi flow Fruetus AurantiiImmaturus(FAI) improving cerebral infarction in acute stage ofgastrointestinal dysfunction;3、To research the mechanism of improving effects of FAI in rats withgastrointestinal dysfunction diseases after cerebral infarction.Methods：We used10weeks old male Wistar rats as the research object, andestablished the middle cerebral artery occlusion model(MCAO)by the lineembolus.80rats were screened after Longa test and randomly divided into5groups:, model group, FAI group, Nimodipine group, FAI+Nimodipine group, andAngelica dahurica group. Another16rats were established into sham operationgroup and16normal rats as normal control group. The above7groups of ratswere randomly divided into24h and4D two subgroups, a total of14groups.Each group had8rats, and they were112rats in all. Immediately awoke aftermodeling, Rats in every group were treated by FAI suspension (6.75mg/ml),Nimodipine suspension (0.9mg/ml), FAI+Nimodipine mixture (containing13.5mgFAI and1.8mg Nimodipine), Angelica dahurica suspension (6.75mg/ml) in properorder respectively, every treatment were2ml liquid; model group, shamoperation group and the normal group was given the same volume of saline.24hsubgroups were treated for just1time and guillotined after24hours;4Dsubgroups were killed after4days of continuous treatment. Rats’ nervous system defect behaviors were observed and recorded for2times, immediatelyafter rats awaking and before their death. We calculated the volume ofinfarction part of brain tissue in rats, observed the pathological changesof gastrointestinal mucosa, and immune-labeled with antibodies againstsubstance P(SP) to quantify nerves, c-kit for interstitial cells of Cajal(ICC), Connexin43(Cx43) for gap-junctions between smooth muscle cells. Tissuewas also examined by transmission electron microscopy (TEM).Results:1、The Longa score results showed that FAI group, nimodipine group andFAI+nimodipine group were significantly lower than model group and Angelicadahurica group (P<0.01). And all the infarction groups showed significantdifferences compared with the sham operation group (P<0.01).2、By staining with TTC, infarction area of brain tissue form all theinfarction groups was observed in white. Calculation the infarct volumeshowed that all the infarction groups had significant difference (P <0.01)with sham operation group. What’s more, the volume of cerebral infarctionwas reduced in order, from rats in FAI group, nimodipine group and FAI+nimodipine group. There was significant difference with the model group andAngelica group (P <0.05).3、There were different degrees of gastric mucosa injury in infarctionrats, which showed significant differences compared with the sham operationgroup (P<0.01).Among them, intervention by FAI and nimodipine cloud improvegastric mucosal injury, in which FAI+nimodipine group was most effective,no obvious difference compared with the sham operation group (P>0.05).Angelica dahurica group was no significant difference compared with modelgroup (P>0.05).4、Pathological mucosa of small intestine score results showed, damageof intestinal mucosa in model group, angelica group, nimodipine group, FAIgroup, FAI+Nimodipine group rats were reduced in order, with significant difference between the sham operation group (P <0.01). Compared with the modelgroup, nimodipine group, FAI group and FAI+nimodipine group had significantdifference (P<0.01).5、Effects of FAI in SP, Cx43, C-kit in gastrointestinal tissue of ratswith acute cerebral infarction.SP, Cx43, C-kit immune positive substancesin gastrointestinal tissues decreased significantly,in infarction groups，compared with the sham operation group (P<0.01). By the intervention of FAIand Nimodipine rats gastrointestinal tissues SP, Cx43, C-kit immune positivematerial expression is higher than the model group (P <0.01), among them FAI+nimodipine group’s effect is most obvious,compared with the sham operationgroup no significant difference (P>0.05). Effect of Angelica dahurica groupis not obvious, and no difference between the model group (P>0.05).6、In normal control group and sham operation group, ICC was spindle-shapedwith a huge ovate nucleolus. A mass of mitochondria, numerous free ribosomes,smooth and rough endoplasmic reticulum were present in the cytoplasm. ICClocated around the nerve fibers, and contacted with neighboring smooth musclecells by gap-junctions. In model group and Angelica dahurica group, thejunctions between ICC and other cells were absent; and the structure of ICCwas unclear with endoplasmic reticulum dilated and threshed, mitochondrialswollen and vacuolated, vacuole formed in cytoplasm, and concave karyothecawiden. In nimodipine group, FAI group, FAI+Nimodipine group, theconfiguration of ICC was almost normal. The mitochondria slightly swelled andthe ridge of mitochondria existed. The rough endoplasmic reticulum lesion wasnot obvious. ICC was normal with intact connections between cells.Conclusion:1、Pathological changes were showed in gastric mucosal morphology of ratswith acute cerebral infarction.2、Changes of gastric interstitial cells of Cajal structure and functionmay be one of mechanism of gastrointestinal dysfunction in acute cerebral infarction.3、Application of FAI on gastrointestinal dysfunction in acute period ofcerebral infarction has certain protective effect in rat.4、Therapeutic effect of FAI on gastrointestinal dysfunction in acuteperiod of cerebral infarction in rats may be related to improvement of gastricinterstitial cells of Cajal structure and function.