温通活血乳膏通过GRP78-PERK通路对糖尿病周围神经病变大鼠雪旺细胞保护机制研究  

Protective Mechanism of Wentong Huoxue Cream(温通活血乳膏)on Schwann Cells in Diabetic Peripheral Neuropathy Rats through GRP78-PERK Pathway

作　　者：刘珊珊 王先敏[2] 严琦 徐利娟 马静[2] 李惠[2] 王晶[3] 马丽[2] LIU Shanshan;WANG Xianmin;YAN Qi;XU Lijuan;MA Jing;LI Hui;WANG Jing;MA Li(The Fourth Clinical Medical College of Xinjiang Medical University,Urumqi 830000,Xinjiang,China;The Affiliated Hospital of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830000,Xinjiang,China;Hospital of Traditional Chinese Medicine Affiliated to Xinjiang Medical University,Laboratory of Respiratory Disease Research,Urumqi 830000,Xinjiang,China)

机构地区：[1]新疆医科大学第四临床医学院,新疆乌鲁木齐830000 [2]新疆医科大学附属中医医院,新疆乌鲁木齐830000 [3]新疆医科大学附属中医医院呼吸病研究重点实验室,新疆乌鲁木齐830000

出　　处：《辽宁中医药大学学报》2025年第5期22-29,F0003,共9页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金(82060842);新疆维吾尔自治区重点研发计划项目(2023B03002)。

摘　　要：目的探讨温通活血乳膏对链脲佐菌素诱导的糖尿病周围神经病变(diabetic peripheral neuropathy,DPN)大鼠坐骨神经结构改变及雪旺细胞凋亡的影响。方法动物实验:取90只雄性SD大鼠随机选择15只作为空白组,余75只造模成功后,将大鼠随机分为5组:模型组、温通活血乳膏高剂量组、温通活血乳膏中剂量组、基质组、二甲双胍组,连续给予温通活血乳膏外涂4周后观察各组大鼠的血糖、热甩尾时间;采用光镜、电镜分别观察大鼠坐骨神经病理变化和超微结构变化;用Western Bolt检测坐骨神经中凋亡相关蛋白Bax与Bcl-2的表达水平;采取TUNEL方法对各组坐骨神经进行检测,观察各组神经细胞凋亡情况。体外实验:选用大鼠雪旺细胞RSC96,分为正常组,高糖组,低、中、高不同剂量的温通活血乳膏组,用Western Bolt及qPCR检测细胞中GRP78、PERK、CHOP、Bax、Bcl-2蛋白表达及mRNA转录水平。结果动物实验:与空白组相比,模型组坐骨神经病理变化和超微结构发生明显改变,坐骨神经中Bax蛋白表达量升高(P>0.05)、Bcl-2蛋白表达量降低(P<0.05),雪旺细胞凋亡数量明显增加;与模型组相比,温通活血乳膏组抑制了DPN大鼠坐骨神经病理和超微结构改变,并逆转了Bax和Bcl-2蛋白表达量(P<0.05),减少了神经髓鞘中细胞凋亡。体外实验:与正常组相比,高糖组GRP78、PERK、CHOP、Bax蛋白表达水平及mRNA转录水平均升高,Bcl-2蛋白表达水平及mRNA转录水平降低(P<0.05);与高糖组相比,温通活血乳膏干预后其表达水平均有不同程度逆转(P<0.05)。结论温通活血乳膏通过GRP78-PERK通路抑制糖尿病周围神经病变大鼠的雪旺细胞凋亡,保护周围神经髓鞘的完整性,从而改善DPN大鼠周围神经结构的损伤。Objective To investigate the effect of Wentong Huoxue Cream(温通活血乳膏)on the structural changes of sciatic nerve and Schwann cell apoptosis in rats with streptozotocin-induced diabetic peripheral neuropathy(DPN).Methods Animal experiment:15 of 90 male SD rats were randomly selected as the blank control group,and the remaining 75 rats were randomly divided into five groups:model group,Wentong Huoxue Cream high-dose group,Wentong Huoxue Cream medium-dose group,matrix group,metformin group,and the blood glucose and hot tail flicking time of the rats in each group were observed after continuous administration of Wentong Huoxue Cream for 4 weeks.Light microscopy and electron microscopy were used to observe the pathological changes and ultrastructural changes of sciatic nerve in rats,respectively.Western Bolt was used to detect the expression levels of apoptosis-related proteins Bax and Bcl-2 in sciatic nerves.The TUNEL method was used to detect the sciatic nerve in each group,and the apoptosis of nerve cells in each group was observed.In vitro experiment:Rat Schwann cells RSC96 were selected and divided into blank group,high glucose group,and low,medium and high doses groups of Wentong Huoxue Cream,and the protein expression and mRNA transcription levels of GRP78,PERK,CHOP,Bax and Bcl-2 in the cells were detected by Western Bolt and qPCR.Results Animal experiments:Compared with those of the blank group,the pathological changes and ultrastructure of sciatic nerve in the model group were significantly changed,the expression of Bax protein in sciatic nerve increased(P>0.05),the expression of Bcl-2 protein decreased(P<0.05),and the number of apoptosis of Schwann cells increased significantly.In vitro experiments:Compared with those of the blank group,the expression levels of GRP78,PERK,CHOP and Bax protein and mRNA transcription levels were increased in the high glucose group,and the expression levels of Bcl-2 protein and mRNA transcription were decreased(P<0.05),and the expression levels of GRP78,PERK,CHOP and B

关 键 词：温通活血乳膏 糖尿病周围神经病变 大鼠 细胞 凋亡 

分 类 号：R255.4[医药卫生—中医内科学]