Study On Anti-inflammatory Effects Of Acupuncture At DU20and ST36in Cerebral Ischemia Reperfusion Injury Rats

Research objectives:The cerebral ischemia reperfusion injury (CIRI) model rats were made by line bolt method. We detected the inflammatory factors of interleukin(IL)-1β, IL-6and tumor necrosis factor(TNF)-a in bilateral brain regions and peripheral serum, as well as the injury stress protein heat shock protein70(HSP70) in peripheral serum, to explore the anti-inflammatory effects of acupuncture at DU20and ST36in cerebral ischemia reperfusion injury rats.Research methods:Rats were randomly divided into normal group, sham group, model group and acupuncture group, the later three groups were further divided into12h,24h,48h,72h,96h and144h six timepoints according to the reperfusion time,8rats in each time group. Middle cerebral artery occlusion (MCAO) was performed in rats. Reperfusion was started at2h after onset of MCAO. Acupuncture group was treated by acupuncturing at acupoints DU20and ST36in required time after successful operation.TTC staining was used for testing model and providing the positioning mark; HE staining was to observe the morphotype of the cells. Immunohistochemistry staining was used for analyzing the expression of IL-1β, IL-6and TNF-α in rats brain tissue; Enzyme-linked immuno sorbent assay (ELISA) was to detect the related inflammation signal and HSP70in peripheral serum. All these techniques were to explore the possible interventional role of acupuncture for the mechanisms of cerebral ischemia reperfusion injury in rats.Research results:1. TTC staining showed that the area of infarction core was in white colour compared with the common red brain tissue. Among the five slices of brain, the slice around optic chiasma area had the most stable infarction region, which provided a good experimental positioning mark to explore the possible anti-inflammatory mechanisms.2. HE staining showed the pathological lesion of infarct core reached the peak at 48h, the cerebral interstitial oedema was alleviated until the72h. Also many glial cells and collagen fibers proliferated. The proliferation of glial cells and collagen fibers was the most obvious up to the96th hour.3. ELISA:1) The expression of IL-1(3in peripheral serum of cerebral ischemia rats exhibited a double-peak, the peak time in model group was at24h and96h, the expression of IL-1β in acupuncture group was lower than model group, and declined significantly at24h; the peak expression of IL-1β in sham group was at24h.2) The expression of IL-6in peripheral serum of sham group exhibited a double-peak, the peak time was at24h and72h; Acupuncture group presented a single-valley profile, the valley time was at48h, and the expression of acupuncture group was lower than model group and sham group; The expression of IL-6in all experimental groups were higher than normal control group.3) The expression of TNF-a in peripheral serum of experimental groups exhibited a single-peak, the peak time in both model group and acupuncture group were at24h, but the expression of acupuncture group was lower than model group; The expression of TNF-α in all experimental groups were higher than normal control group.4) the expression of HSP70in peripheral serum of experimental groups were higher than the normal control group, the peak time in both model group and sham group were at12h, the peak time in acupuncture group was at24h; acupuncture group declined to the base line at72h, and the expression level was lower than the model group and sham group.4. Immunohistochemistry staining:1) The expression of IL-1β in ischemia regions exhibited a double-peak, the peak time in model group was at48h and96h, while acupuncture group was at24h and72h, comparing with model group, the expression of IL-1β in acupuncture group declined obviously, but the level was still above the sham group and the control group.2) The expression of IL-6in ischemic brain exhibited a single-peak profile, the peak time in model group was at96h while in acupuncture group was at72h, comparing with model group, the expression of IL-6in acupuncture group declined obviously, but the level was still above the sham group and the control group.3) The expression of TNF-α in ischemia region exhibit a single-peak, the peak time in both model group and acupuncture group was at72h, the expression of TNF-a in acupuncture group was lower than model group obviously, but above the sham group and the control group; the expression in non-ischemic brain was lower than ischemic brain. Research conclusions:1. In the local of brain ischemia injury, acupuncture could inhibit the expression of inflammatory cell factors related to injury, and promote the expression of inflammatory cell factors related to injury-repair, make the repair-phase arriving earlier, then accelerate the transformation from damage to repair.2. The function of acupuncture at DU20and ST36may work through the intervention of the contralateral injury brain and systemic function adjustment. At the same time reduce the inflammation response of immune system in the local brain and promote the activation of damage repair signal chain.Innovation:CIRI model was established by using right middle cerebral artery occlusion and reperfusion,12h,24h,48h,72h,96h and144h six time points was chosed according to the reperfusion time. We explored the role of acupuncture at DU20and ST36on the expression of inflammatory factors interleukin(IL)-1β, IL-6and tumor necrosis factor(TNF)-a in bilateral brain region and peripheral serum, as well as the injury stress proteins HSP70in peripheral serum. The data showed that the possible pathophysiologic effects of acupuncturing at DU20and ST36in CIRI by regulating the inflammation injury and injury-repair network. The intervention of acupuncture at DU20and ST36on the expression of inflammatory factors of bilateral brain region and peripheral serum together in CIRI rats have not been reported home and abroad.