Department of Traditional Chinese Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100730, China.
Chin J Integr Med. 2013 Jul;19(7):517-23. doi: 10.1007/s11655-013-1506-7. Epub 2013 Jul 2.
To evaluate the effect of serum containing Jinmaitong Capsule (JMT) on apoptosis of Schwann cells (SCs) that are cultured in high glucose at the cellular and molecular levels.
SCs were cultured in Dulbecco's modified Eagle's medium (control group), high glucose (50 mmol/L) medium supplemented with 20% rat serum (HG group), and 50 mmol/L glucose medium supplemented with serum containing JMT (JMT group). SC apoptosis was detected using a terminal deoxynucleotidyl transferase dUTP nick end labeling kit. The expression of Bcl-2 and the caspase-3 p20 subunit in SCs were detected by realtime fluorogenic quantitative polymerase chain reaction and confocal laser scanning microscopy, respectively.
No apoptosis was detected in SCs that were cultured in the control group. The percentage of apoptosis of SCs cultured in the HG group was much higher than that in the control group. The apoptosis of SCs in the JMT group was lower than that in the HG group. Fluorescence intensity of Bcl-2 and the expression of Bcl-2 mRNA in SCs that were cultured in the HG group were much lower than those in the control group and much higher than those in the JMT group (P<0.01). The fluorescence intensity of caspase-3 p20 and the expression of caspase-3 p20 mRNA in SCs that were cultured in the HG group were much higher than those in the control group (P<0.01), and they were remarkably lower in the JMT group (P<0.01).
JMT effectively prevents SC apoptosis that is induced by high glucose. This effect may be because of increased expression of Bcl-2 mRNA and protein and decreased expression of caspase-3 p20 mRNA and protein.
从细胞和分子水平评价金迈通胶囊(JMT)含药血清对高糖培养雪旺细胞(SCs)凋亡的影响。
SCs 分别在含 20%大鼠血清的 DMEM 培养基(对照组)、含 50mmol/L 葡萄糖的 DMEM 培养基(高糖组)和含 JMT 含药血清的 50mmol/L 葡萄糖 DMEM 培养基(JMT 组)中培养。用原位末端标记法(TUNEL)检测 SC 凋亡,实时荧光定量 PCR 和激光共聚焦显微镜分别检测 Bcl-2 和 caspase-3 p20 亚基的表达。
对照组 SC 无凋亡,高糖组 SC 凋亡率明显高于对照组,JMT 组凋亡率低于高糖组。高糖组 Bcl-2 荧光强度和 Bcl-2mRNA 表达均明显低于对照组,高于 JMT 组(P<0.01);高糖组 caspase-3 p20 荧光强度和 caspase-3 p20mRNA 表达均明显高于对照组(P<0.01),JMT 组明显低于高糖组(P<0.01)。
JMT 可有效防治高糖诱导的 SC 凋亡,其作用机制可能与上调 Bcl-2mRNA 和蛋白表达、下调 caspase-3 p20mRNA 和蛋白表达有关。