本文選題：文本挖掘 + 氣陰兩虛��； 參考：《廣州中醫(yī)藥大學(xué)》2016年博士論文

【摘要】：目的：1、探討中醫(yī)藥治療糖尿病腎病的用藥特點(diǎn)；2、觀察氣陰兩虛挾瘀證糖尿病腎病患者血清CXCL12等炎癥因子的表達(dá)情況；3、觀察黃芪甲苷對(duì)2型糖尿病db/db小鼠的腎臟保護(hù)作用。方法：1、文獻(xiàn)研究采用文本挖掘的方法,探索中醫(yī)治療糖尿病腎病的的用藥特點(diǎn)。通過(guò)參考《中藥大辭典》(第二版)、《中華人民共和國(guó)藥典》(增補(bǔ)版)及全國(guó)高等中醫(yī)藥院校規(guī)劃教材《中藥學(xué)》(第二版)建立詞表庫(kù)。登錄中國(guó)知網(wǎng)(CNKI),對(duì)中醫(yī)治療糖尿病腎病理論研究、經(jīng)驗(yàn)總結(jié)、臨床試驗(yàn)相關(guān)文獻(xiàn)進(jìn)行搜索。將搜索到的文獻(xiàn)按程序進(jìn)行篩選并轉(zhuǎn)換格式后建立數(shù)據(jù)庫(kù),以該詞表庫(kù)作為關(guān)鍵詞苦對(duì)相關(guān)文獻(xiàn)中的關(guān)鍵詞進(jìn)行提取并保存。采用Cytoscape軟件對(duì)提取到的關(guān)鍵詞進(jìn)行網(wǎng)絡(luò)分析,總結(jié)其用藥特點(diǎn)。2、臨床觀察參考相關(guān)文獻(xiàn)建立診斷及納排標(biāo)準(zhǔn),對(duì)2015年3月至2016年1月在廣州中醫(yī)藥大學(xué)附屬深圳市中醫(yī)院門診及住院就診的糖尿病腎病患者進(jìn)行篩選,納入符合標(biāo)準(zhǔn)的氣陰兩虛挾瘀型糖尿病腎病患者進(jìn)行觀察。觀察內(nèi)容包括一般資料、生化指標(biāo)水平及相關(guān)炎癥因子的表達(dá)。3、動(dòng)物實(shí)驗(yàn)基于以上兩項(xiàng)研究的結(jié)果,采用中藥黃芪提取物黃芪甲苷對(duì)2型糖尿病db/db小鼠進(jìn)行治療。用藥期間,應(yīng)用代謝籠、血糖儀和動(dòng)物無(wú)創(chuàng)血壓計(jì)測(cè)量小鼠的代謝指標(biāo)、血糖和血壓。治療12周后殺檢小鼠,留取血液和尿液。應(yīng)用全自動(dòng)生化分析儀檢測(cè)小鼠腎功能、血白蛋白等生化指標(biāo)；應(yīng)用ELISA檢測(cè)小鼠尿白蛋白排泄率和腎小管損傷指標(biāo)；應(yīng)用PAS染色劑透射電鏡檢測(cè)腎小球和腎小管病理和超微病理?yè)p傷；采用免疫印跡法(Western Blot)檢測(cè)腎臟內(nèi)P-Akt和P-NF-k B蛋白表達(dá)。結(jié)果：1、文獻(xiàn)研究通過(guò)制定檢索式,檢索CNKI糖尿病腎病相關(guān)文獻(xiàn),共獲得中醫(yī)治療糖尿病腎病理論研究、經(jīng)驗(yàn)總結(jié)及臨床試驗(yàn)相關(guān)文獻(xiàn)共3778篇。根據(jù)篩選標(biāo)準(zhǔn),經(jīng)篩選后得到目標(biāo)文獻(xiàn)理論研究文獻(xiàn)58篇,經(jīng)驗(yàn)總結(jié)文獻(xiàn)264篇,臨床試驗(yàn)文獻(xiàn)618篇。將這些文獻(xiàn)轉(zhuǎn)換格式后進(jìn)行關(guān)鍵詞提取,得到關(guān)鍵的一維頻次及二維頻次結(jié)果,并利用Cytoscape軟件對(duì)提取結(jié)果進(jìn)行網(wǎng)絡(luò)關(guān)系分析。在理論研究文獻(xiàn)中,使用頻次處于前三位的中藥方劑為腎氣丸、生脈散、金匱腎氣丸,使用頻次處于前三位的中藥為黃芪、丹參、茯苓。網(wǎng)絡(luò)分析結(jié)果發(fā)現(xiàn)理論研究中,黃芪、丹參處于整體用藥的核心位置,與其他藥物連系最多。經(jīng)驗(yàn)總結(jié)文獻(xiàn)中方劑應(yīng)用居于前三位的是真武湯、六味地黃丸、五苓散,網(wǎng)絡(luò)分析結(jié)果發(fā)現(xiàn)真武湯與前十位中其他方劑幾乎均存在連系,而濟(jì)生腎氣丸、金匱腎氣丸、六味地黃丸、生脈散關(guān)系密切,而其他方劑相對(duì)獨(dú)立。單味中藥挖掘結(jié)果前三位是黃芪、丹參、地黃,網(wǎng)絡(luò)分析結(jié)果發(fā)現(xiàn)黃芪、地黃位于藥物應(yīng)用的中心位置。臨床試驗(yàn)文獻(xiàn)中方劑挖掘結(jié)果前三位分別為六味地黃丸、補(bǔ)陽(yáng)還五湯、真武湯,網(wǎng)絡(luò)分析結(jié)果金匱腎氣丸、六味地黃丸處于核心位置。單位中藥挖掘結(jié)果前三位為黃芪、丹參、地黃,網(wǎng)絡(luò)分析結(jié)果發(fā)現(xiàn)黃芪仍然處于用藥的核心位置。2、臨床觀察研究共納入2015年3月至2016年1月在廣州中醫(yī)藥大學(xué)附屬深圳市中醫(yī)院門診及住院病例共80例,并納入在體檢門診的21例健康人群作為對(duì)照組。所有納入病例分為正常組(N組)21例、糖尿病組(DM組)19例、糖尿病腎病1-2期組(DKD1-2組)19例、糖尿病腎病3期組(DKD3組)20例、糖尿病腎病4期組(DKD4組)21例、糖尿病腎病5期組(DKD5組)21例。各組病例在年齡、性別、體重等基礎(chǔ)資料上差異無(wú)統(tǒng)計(jì)學(xué)意義。但BMI、血壓、血糖及糖化血紅蛋白指數(shù)方面N組明顯低于DN各組。腎功方面,SCr由N組至DN5組患者逐漸升高,BUN含量DN4、5組含量最高,GFR水平在DKD早期升高,后期逐漸下降,末期時(shí)將至最低。脂代謝方面除HDL外,各組結(jié)果較為一致,N組的HDL水平明顯高于其他各組。CXCL12檢測(cè)結(jié)果,N組與DKD5組含量最高,DM組含量最低,且與N組差異具有統(tǒng)計(jì)學(xué)意義。MCP-1在DKD4、5組含量最高,且與N組比較差異具有統(tǒng)計(jì)學(xué)意義。TGF-β1檢測(cè)結(jié)果顯示DKD各組均高于N組。CRP記過(guò)顯示,N組與DKD5組含量最低,而其他DKD各組與N組差異均具有統(tǒng)計(jì)學(xué)意義。3、動(dòng)物實(shí)驗(yàn)動(dòng)物實(shí)驗(yàn)將db/db小鼠隨機(jī)分為模型組和治療組,每組8只,治療組采用黃芪甲苷給藥,持續(xù)12周。8只db/m小鼠作為對(duì)照組。模型組與治療組體重明顯高于正常組,且模型組體重增加快于治療組。實(shí)驗(yàn)結(jié)束時(shí),模型組血壓低于正常組,而治療組血壓明顯升高,且明顯高于模型組,其差異具有統(tǒng)計(jì)學(xué)意義。治療組心率降低,與模型組相比其差異具有統(tǒng)計(jì)學(xué)意義。模型組、治療組小鼠左、右腎重高于正常值,模型組與正常組差異具有統(tǒng)計(jì)學(xué)意義,而與治療組相比差異無(wú)統(tǒng)計(jì)學(xué)意義。在血糖及糖化血紅蛋白方面,模型組與治療組都遠(yuǎn)高于正常組,但治療組與模型組差異無(wú)統(tǒng)計(jì)學(xué)意義。小鼠UAE方面,模型組、治療組均高于正常組。治療組尿白蛋白含量一直低于模型組,且在第8、12周時(shí)與模型組相比,兩者差異具有統(tǒng)計(jì)學(xué)意義。生化指標(biāo)方面,除SCr、TG、LDL外,其余各項(xiàng)指標(biāo)模型組均高于正常組,且差異具有統(tǒng)計(jì)學(xué)意義。而治療組除TC、HDL外,其余各項(xiàng)結(jié)果與模型組相比,差異均具有統(tǒng)計(jì)學(xué)意義。病理改變方面,模型組小鼠腎小球、腎小管肥大,FPW增加、GBM和TBM增厚,與正常組相比其差異均具有統(tǒng)計(jì)學(xué)意義。治療組可減低腎小球、腎小管面積,抑制GBM、 TBM增厚。足突寬度方面,模型組最高,與正常組比較,遠(yuǎn)高于正常組。治療組低于模型組,與模型組差異有統(tǒng)計(jì)學(xué)意義。通路蛋白方面,p-Akt結(jié)果顯示模型組與正常組、治療組比較其差異具有統(tǒng)計(jì)學(xué)意義。p-NF-k B p65的表達(dá)結(jié)果與p-Akt一致。ELISA方面,db/db模型組尿中TGF-β1表達(dá)遠(yuǎn)高于正常組,其與治療組差異具有統(tǒng)計(jì)學(xué)意義。小鼠腎臟TGF-β1治療組遠(yuǎn)低于正常組和模型組,且差異具有統(tǒng)計(jì)學(xué)意義。NGAL結(jié)果顯示,模型組小鼠中含量最高,治療組低于模型組,其差異具有統(tǒng)計(jì)學(xué)意義。KIM-1結(jié)果模型組高于治療組,差異具有統(tǒng)計(jì)學(xué)意義。結(jié)論：1、中藥黃芪是中醫(yī)治療DKD的核心用藥之一；2、DKD患者的發(fā)病與代謝紊亂、血流動(dòng)力學(xué)異常等關(guān)系密切。氣陰兩虛挾瘀型DKD患者血清中炎癥因子表達(dá)增加且增加程度與疾病發(fā)展進(jìn)程有關(guān)；3、黃芪的單體提取物黃芪甲苷對(duì)2型糖尿病db/db小鼠腎臟具有保護(hù)作用。黃芪甲苷可能通過(guò)抑制db/db小鼠腎內(nèi)Akt/NF-k B活化,減輕腎臟損傷,降低db/db小鼠的尿白蛋白,起到腎臟保護(hù)作用。
[Abstract]:Objective: 1, to explore the characteristics of traditional Chinese medicine in the treatment of diabetic nephropathy; (2) to observe the expression of serum CXCL12 and other inflammatory factors in diabetic nephropathy patients with Qi Yin deficiency and stasis syndrome; 3, observe the renal protective effect of Astragaloside on db/db mice of type 2 diabetes. Method: 1, the literature study used the method of text mining to explore the treatment of traditional Chinese medicine. The characteristics of the drug use of diabetic nephropathy. Through the reference to the Chinese Dictionary of Chinese medicine (Second Edition), the People's Republic of China Pharmacopoeia (supplement Edition) and the national medicine colleges and universities, the Second Edition (Chinese pharmacy > (Second Edition), set up a thesaurus library. Logon to the Chinese CNKI, the theoretical study on the treatment of diabetic nephropathy by Chinese medicine, the summary of experience and the related literature of clinical trials. The search is searched. After screening and converting the searched documents according to the program, the database is set up. The key words in the related literature are extracted and preserved by the word list as the key words. Cytoscape software is used to analyze the extracted key words and summarize the characteristics of the medicine.2. The clinical observations refer to the related literature. Screening of diabetic nephropathy patients from March 2015 to January 2016 at the Shenzhen Chinese Traditional Medical Hospital affiliated to Guangzhou University of Chinese Medicine and hospitalized in the affiliated Shenzhen Chinese Traditional Medical Hospital of Guangzhou University of Chinese Medicine. The observation was carried out in the patients with deficiency of Qi Yin deficiency and stasis type diabetic nephropathy. The contents included general data, biochemical index level and related inflammation. The expression of factor.3, based on the results of the above two studies, using Astragalus membranaceus glycoside of Astragalus membranaceus to treat type 2 diabetic db/db mice. During the period of drug use, the metabolic index, blood glucose and blood pressure of mice were measured by the metabolic cage, blood glucose meter and animal noninvasive blood pressure meter. After 12 weeks, the mice were killed and the blood and urine were left. A full automatic biochemical analyzer was used to detect the renal function of mice, serum albumin and other biochemical indexes. The urinary albumin excretion rate and renal tubule injury index were detected by ELISA. The pathological and ultrastructural pathological damage of glomeruli and renal tubules were detected by transmission electron microscopy with PAS stain, and P-Akt and P in kidney were detected by Western Blot The expression of -NF-k B protein. Results: 1. The literature study was made by formulating the retrieval formula and retrieving the related literature of CNKI diabetic nephropathy. A total of 3778 articles related to the theoretical study of diabetic nephropathy, experience summary and clinical trials were obtained. According to the screening criteria, 58 articles were obtained from the theoretical study of target literature and 264 articles were summarized by experience. 618 articles in clinical trial. The key key one-dimensional frequency and two dimensional frequency results were obtained after the conversion of these documents. The results were analyzed by Cytoscape software. In the theoretical study literature, the traditional Chinese medicine prescription in the first three places was used as kidney qi pill, Shengmai Powder and Jingui kidney qi pill. The results of the network analysis showed that Astragalus membranaceus and Salvia miltiorrhiza were in the core position of the whole drug use in the theoretical study, and the best use of Chinese medicine in the first three was Zhenwu soup, six flavour rehmannia pill, five Ling powder, and network analysis results found Zhenwu soup. Almost all the other prescriptions in the first ten places are connected, while Jisheng Shenqi Pill, Jinkui kidney qi pill, six flavored rehmannia pill, and the other prescription are relatively independent. The first three of the results of single Chinese medicine mining are Astragalus membranaceus, Salvia miltiorrhiza, and Rehmannia glutinosa, and Rehmannia glutinosa is located at the center of the drug application. The first three places were six flavors of rehmannia pill, Buyang return five soup, Zhenwu Decoction, zhenkui Shenqi Pill and six flavour Rehmannia Pill in the core position. The three place of the results of traditional Chinese medicine excavation were Astragalus membranaceus, Salvia miltiorrhiza and Rehmannia glutinosa. The results of network analysis found that Astragalus was still at the core of drug use,.2, and clinical observation and research included 201 A total of 80 cases of outpatient and hospitalized cases in Shenzhen Chinese Traditional Medical Hospital of Guangzhou University of Chinese Medicine from March to January 2016 were included, and 21 healthy people were included in the medical check-up clinic as control group. All the cases were divided into normal group (group N) 21 cases, diabetes group (Group DM) 19 cases, diabetic nephropathy 1-2 group (group DKD1-2) 19 cases, diabetic nephropathy 3 phase group (DKD3 There were 20 cases, 21 cases of diabetic nephropathy (DKD4) group (Group 4), 5 stage diabetic nephropathy group (group DKD5) 21 cases. There was no significant difference in age, sex, weight and other basic data in each group. But BMI, blood pressure, blood sugar and glycosylated hemoglobin index were significantly lower in group N than in DN. SCr from group N to DN5 group gradually increased, BUN contained The content of DN4,5 group was the highest, and the level of GFR increased in the early stage of DKD, gradually decreased at the later stage and was the lowest at the end of the stage. The result of lipid metabolism except HDL was the same. The HDL level in N group was significantly higher than that of other.CXCL12 tests. The content of N and DKD5 groups was the highest, and the DM group was the lowest, and the difference between the N group and the N group was statistically significant. The 5 groups had the highest content, and compared with the N group, the difference of.TGF- beta 1 showed that all the DKD groups were higher than the N group.CRP, and the N group and the DKD5 group were the lowest, while the other DKD groups and the N group had statistical significance.3. The animal experimental animal experiment randomly divided the db/db rats into the model group and the treatment group, with 8 rats in each group. The group was administered with astragaloside, and only db/m mice were used as control group for 12 weeks. The weight of the model group and the treatment group was significantly higher than that of the normal group, and the weight of the model group was increased faster than the treatment group. The blood pressure of the model group was lower than the normal group at the end of the experiment. The blood pressure of the model group was significantly higher than that in the treatment group, and the treatment group was significantly higher than the model group, and the difference was statistically significant. The treatment group was significantly higher than the model group. Compared with the model group, the left and right kidney weight of the model group was higher than that of the normal group. The difference between the model group and the normal group was statistically significant, but there was no significant difference between the model group and the normal group. In the blood sugar and glycated hemoglobin, the model group and the treatment group were far higher than the normal group, but the model group and the treatment group were far higher than the normal group. There was no significant difference between the treatment group and the model group. The UAE, the model group and the treatment group were all higher than the normal group. The urine albumin content in the treatment group was lower than the model group, and the difference was statistically significant compared with the model group at week 8,12. The biochemical indexes, except SCr, TG and LDL, were all higher than the normal group. The difference was statistically significant in the treatment group except TC and HDL, and the other results were statistically significant compared with the model group. In the pathological changes, the glomerulus, renal tubule hypertrophy, FPW increased, GBM and TBM thickening in the model group, and the difference was statistically significant compared with the normal group. The treatment group could reduce the glomerular and kidney. The area of canaliculus, inhibition of GBM, TBM thickening. The width of the foot process was the highest in the model group. Compared with the normal group, it was far higher than the normal group. The treatment group was lower than the model group. The difference between the model group and the model group was statistically significant. The p-Akt results showed that the model group was compared with the normal group, and the treatment group had a statistically significant difference in.P-NF-k B p65 expression. The TGF- beta 1 expression in db/db model group was much higher than that of the normal group, and the difference of TGF- beta 1 in the model group was significantly higher than that in the treatment group. The TGF- beta 1 group in the mice kidney was far lower than the normal group and the model group, and the difference was statistically significant.NGAL results showed that the content of the model group was the highest, the treatment group was lower than the model group, and the difference between the model group and the model group was lower. The statistical significance.KIM-1 result model group is higher than the treatment group, the difference has statistical significance. Conclusion: 1, Astragalus membranaceus is one of the core drugs for the treatment of DKD in traditional Chinese medicine; 2, the incidence of DKD patients is closely related to metabolic disorders and hemodynamic abnormalities. The expression of inflammatory factors in serum of patients with Qi Yin deficiency and blood stasis type DKD increases and increases. 3, Astragalus membranaceus, the single extract of Astragalus membranaceus, has protective effect on the kidney of type 2 diabetic db/db mice. Astragaloside may inhibit the activation of Akt/NF-k B in the kidney of db/db mice, reduce renal damage and reduce the urinary albumin in db/db mice, and play the role of renal protection.
【學(xué)位授予單位】：廣州中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R259

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7 姜e

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