【摘要】：目的:1.動(dòng)物實(shí)驗(yàn)部分:探討糖尿病腎病大鼠腎組織內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)分子GRP78,Caspase-12、CHOP水平及祛痰通絡(luò)湯的干預(yù)治療效果及機(jī)制.2.臨床實(shí)驗(yàn)部分:臨床觀察祛痰通絡(luò)湯對(duì)糖尿病腎病患者的療效。方法:1.動(dòng)物實(shí)驗(yàn)部分:糖尿病腎病模型的制備采用高糖高脂飼料喂養(yǎng)大鼠,并腹腔注射STZ的方法。隨機(jī)分為正常對(duì)照組、模型組、祛痰通絡(luò)湯組、四苯基丁酸組,祛痰通絡(luò)湯組大鼠按2m L/100g劑量灌胃,4-PBA組大鼠按1g/kg的劑量每日灌胃,治療8周。檢測生化指標(biāo)如糖化血紅蛋白、血肌酐、尿素氮等指標(biāo),觀察大鼠腎臟組織的病理改變;應(yīng)用免疫組織化學(xué)和Western-blot檢測糖尿病腎病大鼠腎組織內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)蛋白GRP78、Caspase-12、CHOP的表達(dá).2.臨床實(shí)驗(yàn)部分:84例糖尿病腎�、�-Ⅳ期痰瘀阻絡(luò)證患者隨機(jī)分為治療組42例,對(duì)照組42例,干預(yù)2個(gè)月,觀察患者尿白蛋白排出率(UAE)、尿素氮、肌酐、胱抑素C(Cys C)、糖化血紅蛋白的變化。結(jié)果:動(dòng)物實(shí)驗(yàn)部分1.模型組血糖、糖化血紅蛋白水平明顯升高,治療組血糖、糖化血紅蛋白水平較模型組明顯下降(P0.05),祛痰通絡(luò)湯組下降更明顯(P0.05)。2.與對(duì)照組相比,糖尿病腎病大鼠模型組尿素氮、肌酐、胱抑素C水平明顯升高(P0.05);與模型組相比,治療組的尿素氮、肌酐、胱抑素C水平明顯下降(P0.05),祛痰通絡(luò)湯組治療效果優(yōu)于4-PBA組.3.治療前,模型組及治療組的24h尿蛋白定量無明顯差別(P0.05),治療4周、8周后,祛痰通絡(luò)湯組24h尿蛋白量較模型組及治療前均明顯下降(P0.05).4.HE染色光鏡觀察可見對(duì)照組大鼠腎臟的腎小球結(jié)構(gòu)正常,無毛細(xì)血管基底膜增厚,系膜細(xì)胞無增生,系膜區(qū)無增寬,腎小管結(jié)構(gòu)正常;糖尿病腎病大鼠模型組腎臟腎小球明顯肥大,基底膜明顯增厚,腎小球的系膜細(xì)胞增生,系膜區(qū)增寬,部分腎小管上皮細(xì)胞出現(xiàn)肥大、空泡樣變性;祛痰通絡(luò)湯組及4-PBA組的腎臟病理改變較模型組明顯減輕,祛痰通絡(luò)湯組腎小球病變程度輕于4-PBA組.5.免疫組化染色:5.1免疫組化發(fā)現(xiàn)內(nèi)質(zhì)網(wǎng)分子伴侶GRP78在模型組的表達(dá)明顯高于對(duì)照組,表明ERS在糖尿病腎腎病的腎組織中激活,祛痰通絡(luò)湯降低GRP78蛋白表達(dá),Western-blot的檢測結(jié)果與免疫組化結(jié)果相一致。5.2應(yīng)用Western-blot對(duì)Caspase-12、CHOP的蛋白表達(dá)水平研究,發(fā)現(xiàn)糖尿病腎病大鼠模型組Caspase-12、CHOP的蛋白表達(dá)水平明顯升高(P0.05),與免疫組化結(jié)果結(jié)果一致,免疫組化發(fā)現(xiàn),在模型組大鼠腎臟中的Caspase-12、CHOP的分布范圍和表達(dá)強(qiáng)度明顯高于對(duì)照組,祛痰通絡(luò)湯降低蛋白表達(dá)水平及范圍.臨床實(shí)驗(yàn)部分1.祛痰通絡(luò)湯改善糖尿病腎病患者的臨床癥狀(P0.05)。2.祛痰通絡(luò)湯治療組臨床綜合療效92.5%,明顯優(yōu)于對(duì)照組75%;治療組的血清BUN、SCr、Cys C水平下降,療效明顯優(yōu)于對(duì)照組(P0.05)。結(jié)論:1.糖尿病腎病大鼠模型組腎組織的內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)分子GRP78、Caspase12、CHOP升高,提示糖尿病腎病大鼠腎組織內(nèi)質(zhì)網(wǎng)應(yīng)激激活。2.祛痰通絡(luò)湯能夠降低內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)分子GRP78、Caspase12、CHOP的表達(dá),減輕腎臟病理學(xué)改變,延緩糖尿病腎病的進(jìn)程。3.祛痰通絡(luò)湯能夠降低糖尿病腎病大鼠的血糖、血清肌酐、尿素氮,胱抑素C,減少尿蛋白排出,改善糖尿病腎病大鼠腎功能指標(biāo)。4.祛痰通絡(luò)湯能夠改善糖尿病腎病患者臨床癥狀,改善實(shí)驗(yàn)室檢查指標(biāo),其總有效率達(dá)92.5%,明顯優(yōu)于對(duì)照組(75.0%)。
[Abstract]:Objective: 1. The experimental part of the animal: To study the effect and mechanism of the intervention of the related molecular GRP78, Caspase-12, CHOP, and the expectorant in the kidney of diabetic nephropathy rats. Clinical experiment part: The clinical observation of the effect of the phlegm-removing and collateral-dredging decoction on the patients with diabetic nephropathy. Method: 1. Animal experiment part: The preparation of diabetic nephropathy model adopts high-sugar high-fat feed to feed the rat, and the method of STZ is injected into the abdominal cavity. The rats were randomly divided into the normal control group, the model group, the phlegm-eliminating and collateral-dredging decoction group, the tetraphenylbutyrate group, the phlegm-eliminating and collateral-dredging decoction group, the rats were given intragastric administration at a dose of 2 m L/ 100 g, and the 4-PBA group rats were given intragastric administration at a dose of 1 g/ kg for 8 weeks. The expression of protein GRP78, Caspase-12 and CHOP in the renal tissue of rats with diabetic nephropathy was detected by immunohistochemistry and Western-blot. The clinical experiment part: 84 patients with type 鈪，

本文編號(hào)：2380059