本文選題：糖尿病性腎病　切入點(diǎn)：腎小球纖維化　出處：《中國(guó)藥理學(xué)與毒理學(xué)雜志》2017年06期

【摘要】：目的探討糖尿病腎小球細(xì)胞煙酰胺磷酸核糖轉(zhuǎn)移酶(Nampt)與骨形態(tài)發(fā)生蛋白7(BMP7)表達(dá)的關(guān)系及煙酰胺單核苷酸(NMN)緩解糖尿病腎小球細(xì)胞炎癥纖維化的作用機(jī)制。方法 (1)動(dòng)物實(shí)驗(yàn):C57/BL6自發(fā)性糖尿病小鼠和C57/BL6野生型小鼠,均采取普通飼料喂養(yǎng),當(dāng)自發(fā)性糖尿病小鼠血糖(34.2±1.9)mmol·L~(-1)并出現(xiàn)明顯腎組織損傷時(shí),取腎組織進(jìn)行病理切片,免疫共聚焦法檢測(cè)腎小球細(xì)胞Nampt、核轉(zhuǎn)錄因子κB p65(NF-κB p65)、沉默調(diào)節(jié)蛋白1(SIRT1)和BMP7的表達(dá)。(2)細(xì)胞實(shí)驗(yàn):葡萄糖200 mmol·L~(-1)培養(yǎng)大鼠腎小球系膜HBZY~(-1)細(xì)胞,在不同時(shí)間(24,48和72 h)以及不同濃度NMN(50,100和200μmol·L~(-1))處理24 h時(shí)后,免疫印跡法檢測(cè)Nampt和BMP7的表達(dá);葡萄糖200 mmol·L~(-1)處理HBZY~(-1)細(xì)胞96 h,免疫熒光法檢測(cè)NF-κB p65和α-平滑肌肌動(dòng)蛋白(α-SMA)的表達(dá);應(yīng)用NMN 100μmol·L~(-1)和Nampt特異抑制劑FK866 10μmol·L~(-1)作用HBZY~(-1)細(xì)胞24 h后,免疫印跡法檢測(cè)HBZY~(-1)細(xì)胞Nampt,BMP7和NF-κB p65表達(dá)。結(jié)果 (1)動(dòng)物實(shí)驗(yàn):自發(fā)性糖尿病小鼠腎小球明顯萎縮,腎小球細(xì)胞Nampt和NF-κB p65的熒光強(qiáng)度比野生型小鼠明顯升高(P0.05),而BMP7和SIRT1的熒光強(qiáng)度顯著降低(P0.01)。(2)細(xì)胞實(shí)驗(yàn):Western蛋白印跡檢測(cè)顯示,葡萄糖200 mmol·L~(-1)培養(yǎng)48和72 h,HBZY~(-1)細(xì)胞Nampt表達(dá)增加(P0.01),BMP7表達(dá)下降(P0.01,P0.05)。葡萄糖200 mmol·L~(-1)條件下加NMN 50,100和200μmol·L~(-1)作用24 h,各組BMP7表達(dá)均增加(P0.01);免疫熒光結(jié)果顯示,與細(xì)胞對(duì)照組比較,葡萄糖200 mmol·L~(-1)處理HBZY~(-1)細(xì)胞,NF-κB p65和α-SMA的熒光強(qiáng)度升高(P0.01);NMN干預(yù)后,與葡萄糖200 mmol·L~(-1)處理組比,Nampt和NF-κB p65表達(dá)降低(P0.01),BMP7表達(dá)增加(P0.01);加FK866后,Nampt表達(dá)降低(P0.01),NF-κB p65表達(dá)下降,BMP7表達(dá)雖然有上升趨勢(shì),但其表達(dá)增高沒有NMN組明顯。結(jié)論嚴(yán)重糖尿病狀態(tài)下,通過抑制內(nèi)源性Nampt過表達(dá)能夠上調(diào)BMP7,從而緩解腎小球細(xì)胞炎癥纖維化作用,NMN可能通過干預(yù)Nampt影響細(xì)胞BMP7表達(dá)。
[Abstract]:Objective to investigate the relationship between Namptase and bone morphogenetic protein 7 (BMP7) expression in diabetic glomerular cells and the mechanism of nicotinamide mononucleotide (NMN) in relieving glomerular inflammatory fibrosis in diabetic rats. Animal experiment: C57 / BL6 spontaneous diabetes mice and C57/BL6 wild-type mice, All of them were fed with common diet. When the blood glucose of spontaneous diabetic mice was 34.2 鹵1.9)mmol Ln-1) and obvious renal tissue injury occurred, the renal tissues were taken for pathological section. Immunoconfocal assay was used to detect the expression of Nampt, NF- 魏 B p65, NF- 魏 B p65, silencing regulatory protein 1 (SIRT1) and BMP7 in rat glomerular Mesangial HBZYT-1) cells: glucose 200 mmol / L ~ (-1)). The expression of Nampt and BMP7 was detected by immunoblotting after 24 h treatment with different concentrations of NMN(50100 and 200 渭 mol L ~ (-1), and 96 h with glucose (200 mmol / L ~ (-1)). The expression of NF- 魏 B p65 and 偽 -smooth muscle actin (偽 -SMA-1) was detected by immunofluorescence assay at 96 h after treatment with glucose (200 mmol / L ~ (-1)), and the expression of NF- 魏 B p65 and 偽 -smooth muscle actin (偽 -SMA-1) was detected by immunofluorescence assay. NMN 100 渭 mol L ~ (1) and FK866 10 渭 mol L ~ (-1), a specific inhibitor of Nampt, were used to detect the expression of Nampttsil-BMP7 and NF- 魏 B p65 in HBZY _ (1) cells 24 h after treatment with Nampt specific inhibitor FK866 10 渭 mol L ~ (-1). Results: glomerular atrophy was observed in spontaneously diabetic mice. The fluorescence intensity of Nampt and NF- 魏 B p65 in glomerular cells was significantly higher than that in wild-type mice, while the fluorescence intensity of BMP7 and SIRT1 decreased significantly. After cultured with glucose 200 mmol / L for 48 and 72 h, the expression of Nampt was increased, and the expression of BMP7 was decreased. The expression of NMN 50100 and 200 渭 mol / L ~ (-1) was increased 24 h after the addition of NMN 50100 and 200 渭 mol / L ~ (-1). The results of immunofluorescence showed that, compared with the control group, the expression of BMP7 in each group was significantly higher than that in the control group (P _ (0.01) and P _ (0.01)), and the results of immunofluorescence showed that, compared with the control group, the expression of BMP7 in each group was significantly higher than that in the control group (P _ (0.01) and ~ (-1)). The fluorescence intensity of NF- 魏 B p65 and 偽 -SMA was increased after the treatment of glucose 200 mmol / L ~ (-1)). Compared with glucose 200 mmol L ~ (-1) group, the expression of nmpt and NF- 魏 B p65 decreased, and the expression of P0.01BMP7 increased; after adding FK866, the expression of P0.01- 魏 B p65 decreased, and the expression of P0.01- 魏 B p65 decreased, although the expression of P0.01- 魏 B p65 decreased, although the expression of P0.01- 魏 B p65 decreased, although the expression of P0.01- 魏 B p65 decreased, the expression of P0.01- 魏 B p65 decreased, and the expression of P0.01- 魏 B p65 decreased after treatment with Glucose-200 mmol / L ~ (-1). Conclusion under the condition of severe diabetes mellitus, inhibiting the overexpression of endogenous Nampt can up-regulate the expression of BMP7 and alleviate the inflammatory fibrosis of glomerular cells. NMN may influence the expression of BMP7 by interfering with Nampt.
【作者單位】： 桂林醫(yī)學(xué)院公共衛(wèi)生學(xué)院;桂林醫(yī)學(xué)院生物技術(shù)學(xué)院;桂林醫(yī)學(xué)院藥學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(31060161);國(guó)家自然科學(xué)基金(81460164) 廣西壯族自治區(qū)自然科學(xué)基金(2015GXNSF)~~
【分類號(hào)】：R587.2;R692.9

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