本文選題：艾滋病　切入點：中藥　出處：《成都中醫(yī)藥大學》2016年碩士論文　論文類型：學位論文

【摘要】：目的本研究通過Western Blot技術對前期iTRAQ技術篩選出的艾滋病脾腎陽虛證患者的顯著性差異蛋白進行驗證,以期獲得脾腎陽虛證候的特征性表達蛋白。并進一步予相應中藥干預后觀察蛋白表達變化,探索藥物療效與證候特征性蛋白表達變化之間規(guī)律。方法采集健康對照組、艾滋病脾腎陽虛證患者治療前及治療48周后的外周血,分離血清,提取蛋白質,經(jīng)過蛋白質消化后進行Western Blot檢測。運用imageJ軟件分析灰度值,將每泳道所得出目的蛋白的灰度值與相應的白蛋白灰度值做比較,得出相對灰度值,然后將每種抗體中的正常組的相對灰度值和病人組的相對灰度值導入到GraphPad Prism5軟件。采用IBM SPSS Statistics V20.0軟件進行統(tǒng)計學方法分析。計數(shù)資料采用Fisher精確概率法。正態(tài)分布資料采用均數(shù)±標準差(x±s)表示。用以P0.05標準作為差異性標準。結果1、艾滋病期脾腎陽虛證用藥前與正常對照組比較,RBP4在艾滋病期脾腎陽虛證表達上調,差異有統(tǒng)計學意義(P0.05),與iTRAQ篩選結果一致；C7在艾滋病期脾腎陽虛證表達上調,差異有統(tǒng)計學意義(P0.05),與iTRAQ篩選結果一致。2、無癥狀期脾腎陽虛證用藥前與正常對照組比較,RBP4在無癥狀期脾腎陽虛組表達上調,差異有統(tǒng)計學意義(P0.05),與iTRAQ篩選結果一致,且比艾滋病期蛋白含量低。C7在無癥狀期脾腎陽虛證組表達下調,與iTRAQ篩選結果不一致。3、艾滋病期脾腎陽虛證患者藥物干預后48周后與治療前比較,CD4+、CD8趨于穩(wěn)定升高,CD4具有統(tǒng)計學意義(P0.05),CD8無統(tǒng)計學意義。無癥狀期脾腎陽虛證患者治療后一年與治療前比較,CD4+、CD8趨于穩(wěn)定升高,CD4具有統(tǒng)計學意義(P0.05)。4、艾滋病期脾腎陽虛證用藥前后比較,RBP4在艾滋病期脾腎陽虛證用藥后表達下調,差異有統(tǒng)計學意義(P0.05)。C7在艾滋病期脾腎陽虛證用藥后表達上調。5、無癥狀期脾腎陽虛證用藥前后比較,RBP4在無癥狀期脾腎陽虛組表達下,差異有統(tǒng)計學意義(P0.05),但與艾滋病期相比蛋白含量低。C7在無癥狀期脾腎陽虛證組表達下調。結論1、本研究得出艾滋病脾腎陽虛證存在特征性表達蛋白。RBP4可作為脾腎陽虛證無癥狀期向艾滋病期進展的特征性表達蛋白,可能與其氧化應激、炎癥反應或代謝綜合征相關反應相關。C7可以作為艾滋病期脾腎陽虛證特征性表達蛋白,可能與其免疫反應功能及涉及的細胞溶解和細胞死亡等過程相關。2、本研究顯示RBP4蛋白表達變化水平與艾滋病脾腎陽虛證中藥辨證論治治療療效趨勢呈一致性,對于艾滋病脾腎陽虛證的治療具有療效評價意義。
[Abstract]:Objective in this study, the Western Blot technique was used to verify the differential protein of the patients with deficiency of spleen and kidney yang in AIDS patients who were screened by iTRAQ technique in the early stage. In order to obtain the characteristic expression protein of spleen and kidney yang deficiency syndrome, and to observe the change of protein expression after intervention of corresponding Chinese medicine, to explore the law between the effect of medicine and the change of characteristic protein expression of syndrome. Methods the healthy control group was collected. The peripheral blood of AIDS patients with deficiency of spleen and kidney yang was separated before and 48 weeks after treatment, the serum was isolated, the protein was extracted, the Western Blot was detected after protein digestion, and the gray value was analyzed by imageJ software. Compare the gray value of the target protein with the corresponding gray value of albumin, and get the relative gray value. Then, the relative gray value of normal group and patient group were imported into GraphPad Prism5 software. The statistical method was analyzed by IBM SPSS Statistics V20.0 software. The counting data were counted by Fisher exact probability method. The data were expressed by mean 鹵standard deviation (x 鹵s). P05 standard was used as the difference standard. Results 1. The expression of RBP4 was up-regulated in the patients with deficiency of spleen and kidney yang in the AIDS phase compared with the control group. The difference was statistically significant (P 0.05), which was consistent with the results of iTRAQ screening. The expression of C7 was up-regulated in the syndrome of deficiency of spleen and kidney yang in AIDS stage. The difference was statistically significant (P 0.05), which was consistent with the screening results of iTRAQ. The expression of RBP4 was up-regulated in asymptomatic Spleen-Kidney Yang deficiency group and normal control group before medication, and the difference was statistically significant (P 0.05), which was consistent with that of iTRAQ screening. The protein content was lower than that of AIDS phase. C7 expression was down-regulated in the asymptomatic group with deficiency of spleen and kidney yang. The results of iTRAQ screening were not consistent with the results of iTRAQ screening. 3. There was no significant difference between 48 weeks after drug intervention and before treatment in patients with deficiency of spleen and kidney yang in AIDS. One year after treatment, CD _ 4 and CD _ 8 tended to increase steadily. CD _ 4 was significantly higher than that before treatment. The expression of RBP4 was down-regulated before and after the treatment of deficiency of spleen and kidney yang in AIDS stage. The difference was statistically significant (P 0.05). C7 expression was upregulated after treatment of deficiency of spleen and kidney yang in AIDS stage, and RBP4 expression in asymptomatic group of spleen and kidney yang deficiency was compared before and after treatment of asymptomatic syndrome of spleen and kidney yang deficiency. The difference was statistically significant (P 0.05), but the protein content was lower. C7 expression was down-regulated in the asymptomatic syndrome group of deficiency of spleen and kidney yang. Conclusion 1. This study shows that there is a characteristic expression protein. RBP4 can be used as spleen and kidney syndrome in AIDS. The characteristic expression protein of the progression from asymptomatic phase to AIDS stage of Yang deficiency syndrome, It may be related to oxidative stress, inflammatory reaction or metabolic syndrome related reactions. C7 may be used as the characteristic expression protein of spleen and kidney yang deficiency syndrome in AIDS. This study showed that the expression level of RBP4 protein was consistent with the therapeutic effect of TCM differentiation and treatment of the deficiency of spleen and kidney yang syndrome in AIDS, which may be related to its immunoreactive function and the process of cell dissolution and cell death, etc. In this study, the expression level of RBP4 protein was consistent with the trend of TCM treatment based on syndrome differentiation. For AIDS spleen and kidney yang deficiency syndrome treatment has the curative effect evaluation significance.
【學位授予單位】：成都中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R259

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