糖痛方对高糖诱导RSC96细胞凋亡与增殖的影响

刘晓星，朱晓云，韦茂英，邓岚，李鸣镝

中国中医科学院广安门医院内分泌科，北京 100053

摘要：目的 筛选中药糖痛方的体外作用最佳剂量。方法 采用含 5～125 mmol/L 不同浓度葡萄糖的DMEM培养基培养 RSC96 细胞，检测不同浓度（0.25、0.5、0.75、1.0、1.5、2.0、3.0、5.0 mg/mL）糖痛方高糖培养 RSC96细胞增殖。采用含 100、125 mmol/L 葡萄糖培养基培养 RSC96 细胞 72 h 后，AV/PI 凋亡试剂盒

和流式细胞仪检测细胞凋亡，计算细胞凋亡率；CCK-8法检测不同时点 RSC96 细胞增殖。结果 RSC96 细胞在 100、125 mmol/L 葡萄糖培养基培养下发生凋亡，凋亡率分别为（7.46±0.96）%、（16.53±1.01）%，

与对照组比较差异有统计学意义（P＜0.05）；不同浓度糖痛方可明显改善高糖对RSC96细胞增殖的抑制作用，

24 h起效浓度为 0.25 mg/mL；0.5、1.0、1.5 mg/mL 3个剂量糖痛方可明显抑制高糖诱导的细胞凋亡，与125 mmol/L

高糖组比较，细胞凋亡率显著降低（P＜0.05，P＜0.01）。结论 在 3 个不同剂量中，1.5 mg/mL 糖痛方抑制细胞凋亡作用最佳。

关键词：糖痛方；RSC96细胞；细胞凋亡；细胞增殖

DOI：10.3969/j.issn.1005-5304.2017.10.012

中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2017)10-0049-04

Effects of Tangtong Formula on RSC96 Schwann Cells Apoptosis and Proliferation Induced by High Glucose

LIU Xiao-xing, ZHU Xiao-yun, WEI Mao-ying, DENG Lan, LI Ming-di (Department of Endocrinology, Guang’anmen Hospital, China Academy of Chinese Medical Sciences, Beijing 100053, China)

Abstract: Objective To screen the optimal dose of Tangtong Formula in vitro. Methods RSC96 Schwann Cells were cultivate by DMEM mediums which contains different concentrations of glucose (5–125 mmol/L). The prevention effects of Tangtong Formula at different concentrations (0.25, 0.5, 0.75, 1.0, 1.5, 2.0, 3.0, 5.0 mg/mL) on the proliferation of RSC96 Schwann Cells induced by high glucose were detected. After the RSC96 Schwann Cells were cultivated in 100 mmol/L and 125 mmol/L high glucose mediums for 72 h, the apoptosis of RSC96 Schwann Cells was detected by flow cytometry Annexin V/PI, and the apoptosis rate was calculate; the proliferation situation of RSC96 Schwann cells in different times was detected by CCK-8 method. Results RSC96 Schwann cells were in apoptosis after being intervened by 100 mmol/L and 125 mmol/L high glucose mediums. The apoptosis rates were respectively （7.46±0.96）% and （16.53±1.01）%, with statistical significance compared with control group (P<0.01). Different concentrations of Tangtong Formula could alleviate the inhibitory effect of high glucose on the proliferation of RSC96 Schwann cells, and the threshold concentration of Tangtong Formula in 24 h was 0.25 mg/mL. The concentrations of Tangtong Formula in 0.5 mg/mL, 1.0 mg/mL, and 1.5 mg/mL could inhibit the apoptosis of RSC96 Schwann cells induced by high glucos, compared with 125 mmol/L high glucose group, the apoptosis rate of RSC96 Schwann cells decreased significantly (P<0.05, P<0.01). Conclusion Among three different doses, when the dose of Tangtong Formula is in 1.5 mg/mL, the effects on inhibiting apoptosis are the best.

Key words: Tangtong Formula; RSC96 Schwann cells; apoptosis; proliferation

糖尿病周围神经病变（diabetic peripheral neuropathy， DPN）是糖尿病最为常见的慢性并发症之一，可引起不同程度的感觉和运动神经功能损害，先后出现肢端

基金项目：国家自然科学基金（81473464）

通讯作者：李鸣镝，E-mail：mdlee2009@126.com 感觉异常、肌力减弱甚至肌萎缩等，严重影响患者的生存和生活质量。糖痛方是本院内分泌科林兰教授治疗 DPN 的经验方，前期临床应用疗效较好[1-2]。课题组前期药理研究表明，糖痛方可提高 Bcl-2，降低

Caspase-9、Caspase-3 活性的表达，抑制雪旺细胞凋

亡、促进增殖[3-4]。但糖痛方抑制细胞凋亡、促进增