芒果甙在糖尿病腎病變小鼠腎臟中連結蛋白43之表現影響

The effect of Mangiferin on regulating connexin 43 expression in the kidney of diabetic nephropathy mice

曾瑩嫻(Ying-Sian Zeng)

指導教授 ： 紀宗呈

長榮大學/健康科學學院/醫學研究所/碩士(2014年)

https://doi.org/10.6833/CJCU.2014.00081

摘要

本研究目的在探討芒果甙(Mangiferin)對於糖尿病腎病變的保護作用，觀察芒果甙是否能改善糖尿病腎病變，並且探討其改善作用是否與調控間隙連結蛋白-43(Connexin43)和骨誘導蛋白-7(bone morphogenetic protein-7)的蛋白表現有關。本研究分別以鏈脲佐菌素(Streptozotocin; STZ)誘導糖尿病小鼠，以及不同葡萄糖濃度暴露之腎小管上皮細胞株(NRK52E) ，觀察給予芒果甙之TGF-β(transforming growth factor-β)、Connexin43和BMP-7等蛋白表現的變化。結果顯示在小鼠腎臟的蛋白表現中TGF-β和糖化中期產物受體(RAGE)會隨著誘導STZ給予的週數增加而表現增加；而BMP-7的表現則是隨著時間增加越來越少，但是在Connexin43則是腎病初期的表現量增加，而腎病末期Connexin43的表現則是減少。大鼠腎臟上皮細胞在高糖暴露之後Connexin43和BMP-7的蛋白表現有減少的趨勢，而TGF-β的蛋白表現增加。糖尿病小鼠給予芒果甙預防或者治療之後， 血液尿素氮(BUN)和肌酸酐(Cretinine)均有下降的的情形，在腎臟的病理切片方面，則是可以看到腎絲球肥大及腎小管壞死與細胞間質增生等現象都有改善。芒果甙在高糖暴露之下的腎小管細胞中安全濃度為50 μg/ml，觀察高糖的暴露下Connexin43的表現量會減少，給予芒果甙之後Connexin43的表現增加，芒果甙能抑制高糖所促使的自由基的產生；另外，受到干擾RNA(siRNA)抑制 Connexin43表現的腎小管細胞會增加細胞的死亡並增加自由基的產生，給與芒果甙之後則能減少細胞死亡與降低自由基的產生。芒果甙能抑制因高糖所產生的自由基以及增加Connexin43的表現，而減緩糖尿病腎病變的發生。

關鍵字

芒果甙 ； 糖尿病腎病變 ； 間隙連接蛋白 ； 活性氧

並列摘要

The study aimed to investigate the therapeutic effect of Mangiferin for diabetic nephropathy. We also observed the effect of Mangiferin on regulating Connexin 43 and BMP-7expression in the kidney of diabetic nephropathy mice.In vitro, we used the NRK52E cell to observe the effect of treatment with different glucose concentration . In vivo, we treated the Mangiferin to streptozotocin (STZ)-induced diabetic mice and observed the renal protective effect of Mangiferin as well as analyzed the TGF-β、Connexin43 and BMP-7 protein expression by Western blot.The results show that TGF-β and RAGE expression were increased but BMP-7 and Connexin 43 were decreased in the kidney of STZ mice. High glucose invreased the expression of TGF-β and RAGE but decreased Connexin 43 and BMP-7 protein expression in NRK52E cell. In vivo, Mangiferin administration show both the preventing and therapeutic effect on STZ-diabetic mice, in decreasing the serum levels of blood urea nitrogen (BUN) and Creatinine. We observed glomerular hypertrophy and tubular necrosis in the kidney of STZ mice and Mangiferin treatment could improved that .With the preventing treatment or therapeutic treatment of Mangiferin, the expression of TGF-β and RAGE were decreased while BMP-7 and Connexin 43 were increased in the kidney of STZ mice. We also found that high glucose increased ROS could be decreased by Mangiferin. The cell death and ROS generation of NRK52E cell was increased after RNAi knockdown the Connexin43 expression and Mangiferin could reverse that. In conclusion, Mangiferin could inhibit the progression of diabetic nephropathy by decreasing ROS and increasing the expression of Connexin43.

並列關鍵字

Mangiferin ； Diabetic Nephropathy ； Connexin43 ； ROS

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