Tsai Mang-Hung, Wei I-Hua, Jiang-Shieh Ya-Fen, Jou Ming-Jia, Ko Miau-Hwa, Chen Hui-Min, Wu Ching-Hsiang
Department of Anatomy, College of Medicine, China Medical University, Taichung 404, Taiwan.
Neurosci Res. 2008 Mar;60(3):233-43. doi: 10.1016/j.neures.2007.11.003. Epub 2007 Nov 19.
Hyperglycemia is a well-known factor in reducing nocturnal pineal melatonin production. However, the mechanism underlying diabetes-induced insufficiency of pineal melatonin has remained uncertain. This study was undertaken to examine the structure, innervation and functional activity of the pineal gland in streptozotocin (STZ)-induced diabetes in rats by immunohistochemistry, Western blotting and image analysis. The number of the pinealocytes and the volume of pineal were also estimated using stereologic quantification including the optical fractionator and Cavalieri's method. It has also shown a progressive reduction of the total area of the pineal gland and the nuclear size of pinealocytes beginning at 4 weeks of induced diabetes. Surprisingly, the immunoreactive intensities and protein amounts of serotonin (5-HT) and protein gene product (PGP) 9.5 in the pineal gland were progressively increased from 4 weeks of diabetes. Meanwhile, nerve fibers immunoreactive for PGP 9.5 had disappeared. Diabetes-induced neuropathy was observed in nerve fibers containing tyrosine hydroxylase (TH). The affected nerve fibers appeared swollen and smooth in outline but they showed a distribution pattern, packing density and protein levels comparable to those of the age-matched control animals. Ultrastructural observations have revealed diabetes-induced deformity of Schwann cells and basal lamina, accumulation of synaptic vesicles and deprivation of the dense-core vesicles in the axon terminals and varicosities. The increase in immunoreactivities in 5-HT and PGP 9.5 and shrinkage of pineal gland in the diabetic rats suggest an inefficient enzyme activity of the pinealocytes. This coupled with the occurrence of anomalous TH nerve fibers, may lead to an ineffective sympathetic innervation of the pinealocytes resulting in reduced melatonin production in STZ-induced diabetes.
高血糖是降低夜间松果体褪黑素分泌的一个众所周知的因素。然而,糖尿病导致松果体褪黑素分泌不足的潜在机制仍不明确。本研究旨在通过免疫组织化学、蛋白质印迹法和图像分析,研究链脲佐菌素(STZ)诱导的糖尿病大鼠松果体的结构、神经支配和功能活性。还使用包括光学分割器和卡瓦列里方法在内的体视学定量方法,估计了松果体细胞的数量和松果体的体积。结果还显示,从诱导糖尿病4周开始,松果体的总面积和松果体细胞的核大小逐渐减小。令人惊讶的是,从糖尿病4周开始,松果体中5-羟色胺(5-HT)和蛋白基因产物(PGP)9.5的免疫反应强度和蛋白量逐渐增加。与此同时,对PGP 9.5呈免疫反应的神经纤维消失。在含有酪氨酸羟化酶(TH)的神经纤维中观察到糖尿病性神经病变。受影响的神经纤维外观肿胀且光滑,但它们的分布模式、堆积密度和蛋白质水平与年龄匹配的对照动物相当。超微结构观察显示,糖尿病导致雪旺细胞和基膜畸形、突触小泡积聚以及轴突终末和膨体中致密核心小泡缺失。糖尿病大鼠中5-HT和PGP 9.5免疫反应性的增加以及松果体的萎缩表明松果体细胞的酶活性低下。这与异常TH神经纤维的出现相结合,可能导致松果体细胞的交感神经支配无效,从而导致STZ诱导的糖尿病中褪黑素分泌减少。