本文選題：糖尿病腎病 + 硫化氫��； 參考：《山東大學(xué)》2014年碩士論文

【摘要】：目的 觀察高糖誘導(dǎo)的小鼠足突細胞損傷中胱硫醚-γ-裂解酶(cystathionine-γ-lyase,CSE)、緊密連接蛋白2(ZO-2)、裂孔蛋白(Nephrin)及β-連環(huán)蛋白(β-catenin)的表達變化及硫化氫干預(yù)后ZO-2、Nephrin和β-catenin蛋白水平的變化,探討高糖誘導(dǎo)的足突細胞損傷機制、硫化氫對糖尿病腎臟疾病的作用及其保護機制。 方法 將體外傳代培養(yǎng)的小鼠足突細胞(MPCs)系分為(1)高糖組(HG組)：各組細胞脫血清饑餓24h后,用含3Ommol/L葡萄糖的1640培養(yǎng)基分別刺激0、12、24、48h；(2)正常糖組(NG組)；(3)正常糖+DL-炔丙基甘氨酸(DL-propargylglycine, PPG)組(NG+PPG組)：各組細胞脫血清饑餓24h后,用含不同濃度PPG的1640培養(yǎng)基干預(yù)48h；(4)高糖+NaHS組(HG+NaHS組)：各組細胞脫血清饑餓24h后,用含3Ommol/L葡萄糖和100μmol/L NaHS的1640培養(yǎng)基處理48h。以上各組細胞均用不同濃度的甘露醇將滲透壓調(diào)至3Ommol/L,避免滲透壓的差異對本實驗結(jié)果的影響。處理后,用Western blot檢測各組細胞CSE、ZO-2、Nephrin及β-catenin蛋白水平表達差異。 結(jié)果 1.與高糖Oh組相比,高糖處理12h、24h、48h均顯著降低CSE、Nephrin、ZO-2的表達(P0.05),而β-catenin的水平明顯增高(P0.05),4種蛋白變化均表現(xiàn)出時間依賴性； 2.與正常糖組相比,正常糖培養(yǎng)加不同濃度的PPG可顯著抑制ZO-2、Nephrin的表達(P0.05),顯著提高β-catenin的水平(P0.05),3種蛋白變化呈濃度依賴性； 3.與高糖組相比,高糖誘導(dǎo)的足突細胞加NaHS培養(yǎng)后,ZO-2、Nephrin的表達量有明顯增多(P0.01),相反β-catenin的表達量則明顯降低(P0.01); HG+NaHS組與NG組相比較,ZO-2、Nephrin的表達量仍有明顯下降(P0.01),相反β-catenin的表達量仍有明顯升高(P0.01)。 結(jié)論 本研究結(jié)果提示高糖對足突細胞的損傷可能是通過降低ZO-2表達、過度激活Wnt/β-catenin通路、減少Nephrin的表達量來實現(xiàn)的,ZO-2、β-catenin、Nephrin三種蛋白的變化表現(xiàn)出時間依賴性,同時高糖誘導(dǎo)的足突細胞CSE表達降低,也具有時間依賴性,提示CSE降低引起的內(nèi)源性硫化氫釋放減少可能是足突細胞損傷的重要機制之一。而外源性硫化氫對高糖誘導(dǎo)的足突細胞損傷具有一定保護作用,其機制可能與增加ZO-2表達、抑制Wnt/β-catenin通路,進而減少Nephrin的丟失有關(guān)。
[Abstract]:PurposeTo observe the changes of cystathionine- 緯 -lyase (CSEN), tight junction protein 2ZO-2, rifting protein Nephrinin (Nephrinin) and 尾 -catenin (尾 -catenin) and the changes of ZO-2Nephrin and 尾 -catenin protein levels in mouse podocyte cells induced by high glucose, and the changes of ZO-2Nephrin and 尾 -catenin protein levels after hydrogen sulfide intervention.To investigate the mechanism of high glucose induced injury of podocyte, the effect of hydrogen sulfide on diabetic kidney disease and its protective mechanism.MethodMouse podocyte cells (MPCs1) were divided into high glucose group (HG group) and high glucose group (HG group). The cells in each group were deprived of serum for 24 hours after starvation.The 1640 medium containing 3Ommol/L glucose was used to stimulate the 1640 medium containing 3Ommol/L glucose for 48 h respectively) the normal glucose group was treated with DL-propargyl glycine (DL-propargyl glycine) and the normal glucose group (DL-propargyl glycine): after 24 hours of starvation, the cells in each group were starved for 24 hours after deserting.1640 medium containing different concentrations of PPG was used to intervene for 48 h. HG NaHS group in high glucose NaHS group was treated with 1640 medium containing 3Ommol/L glucose and 100 渭 mol/L NaHS for 48 h.The osmotic pressure was adjusted to 3Ommol / L with different concentrations of mannitol to avoid the effect of the difference of osmotic pressure on the results of the experiment.After treatment, Western blot was used to detect the expression of ZO-2Nephrin and 尾 -catenin protein.Result1.Compared with the high glucose group, the protein expression of Nephrininine ZO-2 in CSE was significantly decreased at 24 h or 48 h after treatment with high glucose, while the level of 尾 -catenin was significantly higher than that in the control group, and the changes of 尾 -catenin were all time-dependent.2.Compared with the normal glucose group, normal glucose culture and different concentrations of PPG could significantly inhibit the expression of ZO-2Nephrin and increase the level of 尾 -catenin in a concentration-dependent manner.3.Compared with the high sugar group,On the contrary, the expression of 尾 -catenin in HG NaHS group was significantly lower than that in NG group, but the expression of 尾 -catenin in HG NaHS group was still significantly lower than that in NG group, but the expression of 尾 -catenin was still significantly increased in HG NaHS group compared with NG group.ConclusionThe results suggest that the changes of ZO-2 and 尾 -catenin in podocyte may be time-dependent by decreasing the expression of ZO-2, over-activating the Wnt/ 尾 -catenin pathway and reducing the expression of Nephrin.At the same time, the decrease of CSE expression in podocyte induced by high glucose is also time-dependent, which suggests that the decrease of endogenous hydrogen sulfide release induced by the decrease of CSE may be one of the important mechanisms of podocyte injury.The exogenous hydrogen sulfide may protect podocyte from high glucose induced podocyte injury by increasing ZO-2 expression, inhibiting Wnt/ 尾 -catenin pathway and reducing the loss of Nephrin.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R587.2;R692

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