本文選題：高血壓病 + 氣虛血瘀證�。� 參考：《廣西中醫(yī)藥大學》2017年碩士論文

【摘要】：目的:為進一步明確高血壓病氣虛血瘀證患者血清誘導(dǎo)人臍靜脈內(nèi)皮細胞株(CRL-1730)凋亡的效應(yīng),并對其機制進行探討,同時觀察補陽還五湯對凋亡細胞的干預(yù)效應(yīng),為高血壓病氣虛血瘀證的臨床防治提供實驗依據(jù)。方法:分別收集高血壓病氣虛血瘀證患者、高血壓病非氣虛血瘀證、健康體檢者的血清及補陽還五湯高、中、低劑量組含藥血清,分別作用于體外培養(yǎng)的人臍靜脈內(nèi)皮細胞株,用ELISA檢測細胞培養(yǎng)上清液凋亡相關(guān)蛋白含量,WB檢測細胞凋亡相關(guān)蛋白表達,倒置相差顯微鏡觀察細胞形態(tài)學,MTT法檢測細胞增殖活性,臺盼藍法檢測活細胞率,并觀察補陽還五湯的保護作用。結(jié)果:1、ELISA檢測結(jié)果:與健康組相比,高血壓病氣虛血瘀證組、高血壓病非氣虛血瘀證組細胞上清液中的β-微管蛋白、Caspase-3、Fas、FasL、Bax、Bax/Bcl-2表達量升高,差異有統(tǒng)計學意義(P0.05或P0.01),高血壓病氣虛血瘀證組、高血壓病非氣虛血瘀證組細胞上清液中的Bcl-2表達量降低,差異有統(tǒng)計學意義(P0.05或P0.01)。另外,與高血壓病氣虛血瘀證組相比,補陽還五湯高、中、低劑量組細胞上清液中的β-微管蛋白、Caspase-3、Fas、FasL、Bax、Bax/Bcl-2表達量降低,差異有統(tǒng)計學意義(P0.05或P0.01),Bcl-2表達量升高,差異有統(tǒng)計學意義(P0.05)。2、WB檢測結(jié)果:與健康組相比,高血壓病氣虛血瘀證組、高血壓病非氣虛血瘀證組細胞中的β-微管蛋白、Caspase-3、Fas、FasL、Bax、Bcl-2、Bax/Bcl-2表達量無統(tǒng)計學差異(P0.05);與高血壓病氣虛血瘀證組相比,補陽還五湯高、中、低劑量組蛋白表達量也無統(tǒng)計學差異(P0.05)。3、由倒置相差顯微鏡觀察發(fā)現(xiàn),高血壓病氣虛血瘀證組存在細胞形態(tài)學改變。4、MTT法檢測結(jié)果:與健康組相比,高血壓病氣虛血瘀證組、高血壓病非氣虛血瘀證組OD值減少,差異有統(tǒng)計學意義(P0.05);與高血壓病氣虛血瘀證組相比,補陽還五湯各劑量組OD值逐漸增加,差異有統(tǒng)計學差異(P0.05)。5、臺盼藍染色法檢測結(jié)果:與健康組相比,高血壓病氣虛血瘀證組和高血壓病非氣虛血瘀證組均有不同程度降低;與高血壓病氣虛血瘀證組相比,補陽還五湯各劑量組活細胞計數(shù)百分比均升高。在光學顯微鏡下觀察臺盼藍各組染色細胞分析,高血壓病氣虛血瘀證組、高血壓病非氣虛血瘀證組染色最多,補陽還五湯各劑量組次之,健康組染色最少。結(jié)論:高血壓病氣虛血瘀證患者血清可以誘導(dǎo)體外培養(yǎng)的人臍靜脈血管內(nèi)皮細胞株的凋亡,補陽還五湯對其凋亡有一定的保護作用,這種保護作用可能與降低β-微管蛋白和凋亡相關(guān)蛋白Bax、Fas、FasL、Caspase-3的表達量有關(guān)。
[Abstract]:Objective: to further clarify the effect of serum induced apoptosis of human umbilical vein endothelial cell line CRL-1730 in patients with qi deficiency and blood stasis syndrome of hypertension, and to explore its mechanism, and to observe the intervention effect of Buyang Huanwu decoction on apoptotic cells.To provide experimental basis for the clinical prevention and treatment of qi deficiency and blood stasis syndrome in hypertension.Methods: the serum of patients with qi deficiency and blood stasis syndrome of hypertension, non-qi deficiency and blood stasis syndrome of hypertension, the serum of healthy persons and the high, middle and low dosage groups of Buyang Huanwu decoction were collected respectively and acted on human umbilical vein endothelial cell lines cultured in vitro.The expression of apoptosis-related protein in supernatant of cell culture was detected by ELISA and WB. The cell proliferation activity was detected by inverted phase contrast microscope and the rate of living cells was detected by trypan blue method.The protective effect of Buyang Huanwu decoction was observed.Results: compared with the healthy group, the expression of 尾 -tubulin (尾 -tubulin) in the supernatant of the cell supernatant of hypertensive patients with qi deficiency and blood stasis syndrome, and the expression of 尾 -tubulin caspase-3, FasLFasLFasLBax-Bax-Bax-Bax-Bax / Bcl-2 was higher than that in the healthy group.The difference was statistically significant (P 0.05 or P 0.01). The expression of Bcl-2 in the supernatant of hypertension non-qi deficiency and blood stasis group was significantly lower than that of hypertension group (P 0.05 or P 0.01).In addition, compared with the group of qi deficiency and blood stasis syndrome of hypertension, the expression of 尾 -tubulin in the supernatant of cell supernatant of Buyang Huanwu decoction was higher than that of the control group, and the expression of 尾 -tubulin in the supernatant of Caspase-3 / Fas-Fas-Fas-Fas-Fas-Fas-Fas-FasLBax-Bax-Bax-Bax / Bcl-2 was significantly lower than that in the group of qi deficiency and blood stasis syndrome of hypertension.There was no significant difference in the expression of 尾 -tubulin caspase-3Fas-Fas-Fas-Fas-Fas-Bax-Bcl-2Bax-Bax / Bcl-2 in the cells of the hypertension group with qi deficiency and blood stasis syndrome, compared with that of the hypertension group, and the expression of 尾 -tubulin in the cells of the hypertension group was not significantly different from that of the hypertension group, and the expression of 尾 -tubulin protein in the cells of the hypertension group was not significantly different from that of the hypertension group, and the expression of 尾 -tubulin protein in the cells of the hypertension group was similar to that of the hypertension group.Buyang Huanwu decoction was high, medium and low dose histone expression had no statistical difference (P0.05N. 3). Observed by inverted phase contrast microscope, it was found that there were cell morphological changes and MTT assay in hypertension Qi deficiency and blood stasis syndrome group: compared with the healthy group,The OD value of hypertension non-qi deficiency and blood stasis syndrome group decreased, the difference was statistically significant (P 0.05), compared with hypertension qi deficiency and blood stasis syndrome group, the OD value of Buyang Huanwu decoction group increased gradually.The results of trypan blue staining: compared with the healthy group, the blood stasis syndrome group and the non-qi deficiency and blood stasis syndrome group were lower than those of the control group, and compared with the hypertension qi deficiency and blood stasis syndrome group, the blood stasis syndrome group was lower than that of the healthy control group, and compared with the healthy group, the blood stasis syndrome group and the blood stasis syndrome group were lower than those in the healthy control group.The percentage of living cells in each dose group of Buyang Huanwu decoction increased.The staining cells of trypan blue groups were observed under optical microscope. The staining was the most in the hypertension group with Qi deficiency and blood stasis syndrome, the most in the hypertension non-qi deficiency and blood stasis group, the second in the Buyang Huanwu decoction group, and the least in the healthy group.Conclusion: the serum of patients with qi deficiency and blood stasis syndrome of hypertension can induce apoptosis of human umbilical vein endothelial cell line cultured in vitro, and Buyang Huanwu decoction has certain protective effect on apoptosis.This protective effect may be related to the reduction of the expression of 尾 -tubulin and apoptosis-related protein Bax-Fas-Fas-FasL- Caspase-3.
【學位授予單位】：廣西中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R259

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