發(fā)布時間：2018-06-12 16:37

  本文選題：腦缺血損傷 + 補陽還五湯　； 參考：《湖南中醫(yī)藥大學(xué)》2013年碩士論文

【摘要】：目的： 觀察大鼠局灶性腦缺血后CyclinD1及CDK2的表達與細胞增殖的關(guān)系及補陽還五湯的干預(yù)作用,從細胞增殖角度探討補陽還五湯的神經(jīng)保護作用機制,為中醫(yī)藥治療腦缺血性神經(jīng)損傷提供新的認識,為其臨床應(yīng)用提供實驗依據(jù)。 方法： 采用大腦中動脈線栓法(MCAO)建立局灶性腦缺血大鼠模型,將成年大鼠隨機分為4組,即空白組,假手術(shù)組,模型組,補陽還五湯組,給予不同處理,分別于7天,14天,21天處死,對大鼠進行神經(jīng)功能缺失癥狀評分,采用TTC法計算其梗死面積,免疫組化法觀察缺血腦組織CyclinD1及CDK2的表達及BrdU標記的細胞新生情況。 結(jié)果： 1.神經(jīng)功能癥狀評分：正常組、術(shù)后2h假手術(shù)組大鼠未見神經(jīng)功能缺損,造模后各手術(shù)組大鼠都存在有明顯神經(jīng)功能缺損；與假手術(shù)組比較,各時間點的模型組神經(jīng)功能障礙評分顯著增加(P0.01)；與模型組比較,用藥7天,14天,21天后補陽還五湯組神經(jīng)功能缺失癥狀評分減低(P0.05)；說明補陽還五湯對大鼠腦缺血損傷具有一定的保護作用。 2.梗死面積比的測定：正常組,假手術(shù)組均未出現(xiàn)梗死灶,與正常組及假手術(shù)組比,模型組大鼠TTC染色顯示梗死灶非常明顯(P0.01),梗死面積比隨時間延長而減少。與模型組相比,用藥7天,14天,21天后,補陽還五湯組可有效地減小缺血后梗死面積(P0.01),梗死面積隨用藥時間的增加而減少。 2. CyclinDl及CDK2的表達：正常組,假手術(shù)組可見少量的CyclinDl及CDK2蛋白的表達,腦缺血后皮層區(qū)與缺血半暗帶區(qū)CyclinDl及CDK2陽性細胞大量表達(P0.01),7天達高峰,隨時間延長有依次減少的趨勢。用藥7天,14天,21天后,補陽還五湯組能顯著增加腦缺血后皮層區(qū)與缺血半暗帶區(qū)CyclinDl及CDK2的表達,較模型組有顯著差別(P0.01),隨時間延長有依次減少的趨勢。 3.BrdU陽性細胞測定：正常組,假手術(shù)組可見少量BrdU陽性細胞,模型組可見BrdU陽性細胞增加,有隨時間增加的趨勢,但是增長緩慢。用藥7天,14天,21天后,補陽還五湯組能顯著增加BrdU陽性細胞的數(shù)量,差異顯著(P0.01),21天達高峰。相鄰切片可見BrdU陽性細胞的表達區(qū)域與CyclinDl、CDK2的表達區(qū)域基本相同。 結(jié)論： 1.大鼠局灶性腦缺血后腦組織因缺血造成嚴重的神經(jīng)功能損傷和腦梗死,補陽還五湯可減輕缺血后的神經(jīng)功能損傷,減少梗死面積。 2.大鼠局灶性腦缺血后皮層區(qū)與缺血半暗帶區(qū)CyclinDl及CDK2的表達增加,補陽還五湯可增加皮層區(qū)與缺血半暗帶區(qū)CyclinDl及CDK2的表達,早期效果明顯。 3.大鼠局灶性腦缺血后BrdU有少量表達,補陽還五湯可顯著增力口BrdU表達,通過調(diào)控CyclinDl、CDK2的表達使神經(jīng)細胞增殖,從而誘導(dǎo)神經(jīng)再生作用,起到神經(jīng)保護作用。
[Abstract]:Objective: to observe the relationship between the expression of CyclinD1 and CDK2 and cell proliferation after focal cerebral ischemia in rats and to explore the neuroprotective mechanism of Buyang Huanwu decoction from the point of view of cell proliferation. To provide a new understanding for the treatment of cerebral ischemic nerve injury with traditional Chinese medicine, and to provide experimental basis for its clinical application. Methods: the rat model of focal cerebral ischemia was established by middle cerebral artery occlusion (MCAO). Adult rats were randomly divided into four groups: blank group, sham operation group, model group and Buyang Huanwu decoction group. The rats were killed on the 7th day, 14th day and 21st day. The neurological deficit symptoms were scored, the infarct size was calculated by TTC method, the expression of CyclinD1 and CDK2 in ischemic brain tissue and BrdU labeled cells were observed by immunohistochemical method. Results: 1. The neurological function symptom score: normal group, 2 hours after operation group rats did not have nerve function defect, each operation group had obvious nerve function defect after modeling, compared with the sham operation group, Compared with the model group, the neurological dysfunction score of the model group increased significantly at each time point, and compared with the model group, the neurological deficit symptom score of the Buyang Huanwu decoction group decreased P0.05 after 7 days and 14 days and 21 days after treatment. It shows that Buyang Huanwu decoction has certain protective effect on cerebral ischemia injury in rats. 2. Compared with normal group and sham operation group, TTC staining of model group showed that the infarct area was very obvious (P 0.01), and the infarct area ratio decreased with time. Compared with the model group, the Buyang Huanwu decoction group could effectively reduce the infarct area after ischemia (P 0.01) and the infarct area decreased with the increase of medication time after 7 days and 14 days and 21 days later. Expression of Cyclin D1 and CDK2: a small amount of Cyclin D1 and CDK2 protein were found in normal and sham operation groups. The positive cells of cyclin D1 and CDK2 in cerebral ischemic cortex and ischemic penumbra reached the peak at 7 days after ischemia. After 7 days, 14 days and 21 days, Buyang Huanwu decoction group could significantly increase the expression of cyclin D1 and CDK2 in cerebral ischemic cortex and ischemic penumbra. Compared with the model group, there was a significant difference between the model group and the model group (P 0.01), and decreased with time. 3. BrdU positive cells were detected in the normal group and sham operation group, and BrdU positive cells were increased in the model group. But growth is slow. After 7 days, 14 days and 21 days, Buyang Huanwu decoction group could significantly increase the number of BrdU positive cells, and the difference reached the peak at 21 days. The expression of BrdU positive cells in adjacent sections was similar to that of Cyclin Dlt1 CDK2. Conclusion: 1. After focal cerebral ischemia, the brain tissue of rats suffered from severe nerve function injury and cerebral infarction due to ischemia. Buyang Huanwu decoction can reduce the nerve function injury and reduce the infarct area after ischemia. The expression of cyclin D1 and CDK2 in cortex and ischemic penumbra was increased after focal cerebral ischemia in rats. Buyang Huanwu decoction could increase the expression of cyclin D1 and CDK2 in cortex and ischemic penumbra. There was a little expression of BrdU after focal cerebral ischemia in rats, and Buyang Huanwu decoction could significantly increase the expression of BrdU in the mouth of rats. By regulating the expression of CyclinDltl CDK2, nerve cells proliferated, thus inducing nerve regeneration and playing a neuroprotective role.
【學(xué)位授予單位】：湖南中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R285.5;R-332;R277.7

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本文編號：2010337