本文選題：腦缺血再灌注 + 醒腦開(kāi)竅針刺法　； 參考：《南京中醫(yī)藥大學(xué)》2016年博士論文

【摘要】：目的：觀察醒腦開(kāi)竅針刺法延長(zhǎng)腦梗死溶栓時(shí)間窗的效應(yīng)；探討醒腦開(kāi)竅針刺法對(duì)缺血再灌注損傷的腦保護(hù)作用的可能機(jī)制。方法：將SD大鼠分為假手術(shù)組、模型組、溶栓組、針刺+溶栓組(針刺組)、藥物+溶栓組(對(duì)照組),以自體血栓栓塞法制作局灶性腦缺血大鼠模型,rt-PA為溶栓手段,以“醒腦開(kāi)竅”針刺法為干預(yù)手段,觀察針刺干預(yù)對(duì)大鼠神經(jīng)行為學(xué)評(píng)分、腦梗死容積百分比、腦含水量、腦出血量以及血腦屏障通透性的影響；在此基礎(chǔ)上,同時(shí)運(yùn)用Real-time PCR和Western blot方法分別檢測(cè)各組大鼠皮層中炎性因子TNF-α、 IL-1β、ICAM-1的mRNA及蛋白表達(dá)情況；并運(yùn)用明膠酶譜法檢測(cè)MMP-2、MMP-9活性變化,最后運(yùn)用Western blot方法檢測(cè)緊密連接相關(guān)蛋白ZO-1、occludin的表達(dá),以探討針刺調(diào)控基質(zhì)金屬蛋白酶網(wǎng)絡(luò)的作用機(jī)制。結(jié)果：1、針刺延長(zhǎng)腦梗死溶栓時(shí)間窗的效應(yīng)研究溶栓治療后腦梗死大鼠神經(jīng)行為學(xué)評(píng)分、腦含水量、腦出血量以及血腦屏障通透性隨著溶栓時(shí)間的延長(zhǎng)而增加(P0.05)；溶栓治療后腦梗死容積百分比與治療前相比并無(wú)明顯變化(P0.05)；針刺以及抑制劑干預(yù)均能降低神經(jīng)行為學(xué)評(píng)分,減少腦梗死體積、腦含水量以及腦出血量,穩(wěn)定血腦屏障通透性(P0.05)；相比抑制劑,針刺在降低神經(jīng)行為學(xué)評(píng)分、減少腦出血量以及穩(wěn)定血腦屏障通透性上更具優(yōu)勢(shì)(P0.05)；在減少腦梗死體積以及腦含水量上兩者無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。2、針刺延長(zhǎng)腦梗死溶栓時(shí)間窗的機(jī)制研究(1)炎性因子的mRNA及蛋白表達(dá)：溶栓治療后腦梗死大鼠TNF-α、IL-1β以及ICAM-1的mRNA及蛋白表達(dá)均升高(P0.05)；針刺和抑制劑干預(yù)均能降低這三種炎性因子的mRNA及蛋白表達(dá)(P0.05)；相比抑制劑,針刺在降低ICAM-1的mRNA及蛋白表達(dá)上更具優(yōu)勢(shì)(P0.05)；而在降低TNF-α、IL-1β的mRNA及蛋白表達(dá)上二者無(wú)統(tǒng)計(jì)學(xué)差異(P0.05)。(2)基質(zhì)金屬蛋白酶活性：溶栓前后比較MMP-2和MMP-9差異具有統(tǒng)計(jì)學(xué)意義(P0.05)；針刺和抑制劑能降低溶栓后MMP-9活性,且針刺效應(yīng)大于抑制劑效應(yīng)(P0.05)；針刺和抑制劑在降低MMP-2活性上與溶栓組比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)(3)緊密連接蛋白表達(dá)：溶栓治療后腦梗死大鼠ZO-1和occludin的蛋白表達(dá)隨著溶栓時(shí)間的延長(zhǎng)而降低(P0.05)；針刺和抑制劑均能提高溶栓后腦梗死大鼠ZO-1和occludin的蛋白表達(dá)(P0.05)；相比抑制劑,針刺提高溶栓后腦梗死大鼠ZO-1和occludin的蛋白表達(dá)更為顯著(P0.05)。結(jié)論：大鼠腦缺血后溶栓治療受到時(shí)間窗的嚴(yán)格限制,超過(guò)時(shí)間窗溶栓會(huì)導(dǎo)致神經(jīng)功能損傷、血腦屏障改變以及出血性轉(zhuǎn)化,且這些影響隨著溶栓時(shí)間的延長(zhǎng)而加重。醒腦開(kāi)竅針刺法和MMPs廣譜抑制劑強(qiáng)力霉素均可通過(guò)改善腦缺血再灌注大鼠神經(jīng)功能損傷、減少溶栓治療后血腦屏障通透性改變以及出血性轉(zhuǎn)化來(lái)實(shí)現(xiàn)延長(zhǎng)腦梗死溶栓時(shí)間窗的效應(yīng),且前者神經(jīng)保護(hù)作用較后者更明顯。此外,醒腦開(kāi)竅針刺法較MMPs廣譜抑制劑強(qiáng)力霉素能更顯著的抑制腦缺血再灌注大鼠皮層相關(guān)炎性因子的mRNA及蛋白表達(dá),繼而抑制MMP-2、MMP-9過(guò)度激活,從而上調(diào)緊密連接相關(guān)蛋白的表達(dá),從調(diào)控MMPs網(wǎng)絡(luò)途徑更好的保護(hù)腦組織。
[Abstract]:Objective: To observe the effect of Xingnao Kai Qiao acupuncture on prolonging the time window of thrombolytic thrombolysis in cerebral infarction and to explore the possible mechanism of the protective effect of the acupuncture on cerebral ischemia and reperfusion injury. Methods: the SD rats were divided into sham operation group, model group, thrombolytic group, acupuncture + thrombolytic group (acupuncture group), drug + thrombolytic group (control group) and autologous thromboembolism. The rat model of focal cerebral ischemia was made in the legal system. Rt-PA was the means of thrombolytic therapy. The effects of acupuncture intervention on the neurobehavioral score of the rats, the percentage of cerebral infarction, the volume of cerebral infarction, the amount of brain water content, the amount of cerebral hemorrhage and the permeability of blood brain barrier were observed. On the basis of this, Real-time PCR and Western BL were used at the same time. Ot method was used to detect the mRNA and protein expression of inflammatory factors TNF- a, IL-1 beta, ICAM-1, and MMP-2 and MMP-9 activity by gelatin zymography. Finally, the Western blot method was used to detect the expression of ZO-1 and occludin of closely linked proteins, in order to explore the effect of acupuncture on the regulation of matrix metalloproteinase network. Results: 1, the effect of acupuncture prolonging thrombolytic time window of cerebral infarction study on neurobehavioral score of rats with cerebral infarction after thrombolytic therapy, brain water content, cerebral hemorrhage and blood brain barrier permeability increased with the prolongation of thrombolytic time (P0.05); the percentage of infarct volume after thrombolytic treatment was not significantly different from that before treatment (P0.0 5) Acupuncture and inhibitor intervention can reduce neurobehavioral score, reduce cerebral infarction volume, brain water content and cerebral hemorrhage, and stabilize blood brain barrier permeability (P0.05). Compared with inhibitors, acupuncture is more advantageous in reducing neurobehavioral score, reducing cerebral hemorrhage and stabilizing blood brain barrier permeability (P0.05); in reducing the brain There was no statistical difference between the infarct volume and the brain water content (P0.05).2. The mechanism of prolonging the thrombolytic time window of cerebral infarction (1) the mRNA and protein expression of inflammatory factors: the expression of mRNA and protein in TNF- a, IL-1 beta and ICAM-1 in rats after thrombolytic therapy increased (P0.05), and the intervention of acupuncture and inhibitor could reduce these three species. MRNA and protein expression (P0.05) of inflammatory factors; compared with inhibitors, acupuncture was more advantageous in reducing mRNA and protein expression in ICAM-1 (P0.05), but there was no statistical difference in the mRNA and protein expression of IL-1 beta in the decrease of TNF- alpha (P0.05). (2) the activity of matrix metalloproteinase: the difference of MMP-2 and MMP-9 before and after thrombolytic was statistically significant. Meaning (P0.05); acupuncture and inhibitor can reduce MMP-9 activity after thrombolytic, and acupuncture effect is greater than inhibitor effect (P0.05); acupuncture and inhibitors have no significant difference (P0.05) (3) close connexin expression in reducing the activity of MMP-2 with thrombolytic group (P0.05): the protein expression of ZO-1 and occludin in cerebral infarction rats after thrombolytic treatment with thrombolytic thrombolysis Prolongation of time (P0.05); acupuncture and inhibitors can improve the protein expression of ZO-1 and occludin (P0.05) in rats with cerebral infarction after thrombolytic infarction (P0.05). Compared with the inhibitor, the protein expression of ZO-1 and occludin in rats with cerebral infarction after thrombolytic treatment is more significant (P0.05). Conclusion: Thrombolytic therapy in rats after cerebral ischemia is strictly limited by the time window. More than the time window thrombolytic can cause nerve function damage, blood brain barrier change and hemorrhagic transformation, and these effects aggravate with the prolongation of thrombolytic time. The wake brain opening acupuncture method and the MMPs broad-spectrum inhibitor, doxycycline can improve the nerve function of cerebral ischemia reperfusion rats and reduce the permeability of blood brain barrier after thrombolytic therapy. The effect of sexual change and hemorrhagic transformation on prolonging the time window of thrombolytic thrombolysis was achieved, and the former was more obvious than the latter. In addition, the effect of Xingnao opening and orifices acupuncture was more significant than MMPs broad-spectrum inhibitor, doxycycline, to inhibit the mRNA and protein expression of cortical related inflammatory factors in cerebral ischemia reperfusion rats, and then inhibit MMP-2, MM P-9 overactivates the expression of tight junction proteins, which can better protect brain tissue by regulating MMPs network.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R245

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