基于PI3K/AKt信号通路探讨枸杞多糖改善糖尿病雄性大鼠生殖系统损伤的作用研究

Diabetes mellitus associated male reproductive system injury is a serious problem to be solved. Our initial findings has indicated that lycium barbarum polysaccharides can effectively ameliorate the testicular tissue damage and spermatogenic dysfunction. Other research has shown that LBP possesses significant antioxidant effect and protective effect on sperm cells from excessive apoptosis caused by oxidative stress through up-regulating the expression of Bcl-2, inhibiting of the expression of Bax and Caspase3. However, researchers reported that PI3K / Akt pathway plays an important role in regulating the apoptosis of spermatogonia cells caused by oxidative stress. Based on the above, we hypothesized that LBP may exert the anti-apoptotic effects by activating PI3K/ Akt signaling pathways. After that, the Bad translocation is inhibited, Casspase-9 is deactivated and NF-κB is activated..This study intends to clarify the protective effect of LBP on reproductive injury and to explore the relationship between its mechanisms and PI3K/Akt mediated opoptosis signaling pathway by establishing in vivo/vitro models of reproductive system injury in the type 2 diabetic rats.The behavioral pharmacology, immunohistochemistry, confocal laser, western-blot, RT-PCR analysis and other methods are employed. So that,the molecular mechanisms of LBP on reproductive injury is revealed. The result of this study will provide the basis of theoretical and experimental evidence for the transformation of Ningxia local resource advantages into economic advantages.

糖尿病性男性生殖系统损伤是目前亟待解决的问题。我们前期研究发现，枸杞多糖对糖尿病小鼠睾丸组织损伤及生精功能障碍有明显改善作用，另有研究表明枸杞多糖有明显抗氧化作用，并通过上调Bcl-2、抑制Bax及Caspase-3表达，保护生精细胞免受氧化应激引起的过度凋亡。PI3K/Akt通路在调节氧化应激引起生精细胞凋亡中起着极为重要作用。基于以上，我们推测枸杞多糖是通过激活PI3K/Akt，抑制 Bad 转位、使Casspase-9失活、活化NF-κB发挥抗生精细胞凋亡效应，从而起到生殖保护作用。.本课题拟通过建立2型糖尿病生殖损伤整体及离体模型，利用行为学、组织学、RT-PCR、Western Blot等方法，明确枸杞多糖对糖尿病性雄性生殖系统损伤的保护作用，探索其作用机制与PI3K/Akt介导的凋亡通路之间的关系。揭示枸杞多糖干预生殖系统损伤的作用机理。为临床治疗糖尿病男性生殖损伤提供新策略。

糖尿病性男性生殖系统损伤是目前亟待解决的问题。我们前期研究发现，枸杞多糖对糖尿病小鼠睾丸组织损伤及生精功能障碍有明显改善作用。PI3K/Akt通路在调节生精细胞凋亡及自噬过程中发挥着极为重要作用。基于以上，我们推测枸杞多糖是通过激活PI3K/Akt调控生精细胞凋亡及自噬效应，从而起到生殖保护作用。..本课题首先通过建立链脲佐菌素诱导糖尿病生殖损伤小鼠模型，利用行为学、组织学、RT-PCR、 Western Blot 等方法，发现枸杞多糖能显著升高糖尿病小鼠附睾精子数量、 精子活力、 精子活率以及睾丸精子头数量；显著增加糖尿病小鼠血清当中 T、 FSH、 L水平；显著减少糖尿病小鼠睾丸组织中MDA 和 ROS 含量，增强睾丸组织中 SOD、 GSH-Px 和 CAT 活性；明显减轻糖尿病小鼠睾丸曲细精管病理性损伤及生殖细胞超微结构病理性变化；显著降低睾丸组织Beclin-1、 LC3I 免疫阳性细胞的表达；显著降低 Beclin-1、 LC3I/LC3II、 Caspase-3、 Bax 蛋白含量同时明显升高 Bcl-2、 p-PI3K、 p-Akt 蛋白的表达；显著抑制糖尿病睾丸组织在中 Beclin-1、 LC3I mRNA 表达。其次，采用高糖诱导的GC-1精原细胞损伤模型，采用CCK8法、流式细胞术及Western Blot等方法，发现枸杞多糖对GC-1精原细胞具有促增殖抑制凋亡作用，升高P-PI3K/PI3K、P-Akt/Akt的比值；对高糖诱导的GC-1损伤也具有明显的保护作用，上调高糖诱导的GC-1精原细胞中P-PI3K和P-Akt的蛋白表达。..基于以上，本课题明确了枸杞多糖对糖尿病性雄性生殖系统损伤的保护作用，揭示其作用机制与 PI3K/Akt/mTOR 信号通路介导的睾丸细胞自噬和凋亡的关系，证明了枸杞多糖干预生殖系统损伤的作用机理。为寻找治疗糖尿病性功能障碍及生育力低下的药物提供实验依据，为我区资源优势转化为经济优势提供了基础理论。