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1.甘肃中医药大学,兰州 730000
2.贵州中医药大学 第二附属医院,贵阳 550000
3.上海中医药大学,上海 200000
陈彦旭,硕士,从事糖尿病及其并发症的中医药防治研究,E-mail:sdzzcyx@126.com
金智生,教授,主任医师,博士生导师,从事糖尿病及其并发症的中医药防治研究,E-mail:jzsgszy@126.com
收稿日期:2021-06-04,
网络出版日期:2022-01-10,
纸质出版日期:2022-03-20
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陈彦旭,金智生,何流等.红芪多糖对高糖培养下雪旺细胞Bcl-2/Caspase-3信号通路的影响[J].中国实验方剂学杂志,2022,28(06):71-78.
CHEN Yan-xu,JIN Zhi-sheng,HE Liu,et al.Effect of Hedysarum Polysaccharides on Bcl-2/Caspase-3 Signaling Pathway of Schwann Cells Cultured in High Glucose[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(06):71-78.
陈彦旭,金智生,何流等.红芪多糖对高糖培养下雪旺细胞Bcl-2/Caspase-3信号通路的影响[J].中国实验方剂学杂志,2022,28(06):71-78. DOI: 10.13422/j.cnki.syfjx.20220694.
CHEN Yan-xu,JIN Zhi-sheng,HE Liu,et al.Effect of Hedysarum Polysaccharides on Bcl-2/Caspase-3 Signaling Pathway of Schwann Cells Cultured in High Glucose[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(06):71-78. DOI: 10.13422/j.cnki.syfjx.20220694.
目的
2
通过观察红芪多糖(HPS)对高糖培养下雪旺细胞(SCs)B细胞淋巴瘤-2(Bcl-2)、半胱氨酸天冬氨酸蛋白水解酶-3(Caspase-3)、Bcl-2相关X蛋白(Bax)信号通路的影响,探讨HPS在治疗糖尿病周围神经病变(DPN)过程中可能的作用机制。
方法
2
选取出生5~7 d的SD乳鼠4只,随机分为正常组、高糖组、HPS+高糖组、
α-
硫辛酸(
α
-LA)+高糖组,从坐骨神经部位提取SCs,放入37 ℃,5% CO
2
培养箱中培养,待细胞达整瓶的80%后,细胞增殖与活性检测(CCK-8)法筛适宜高糖,HPS及
α
-LA干预的实验浓度,蛋白免疫印迹法(Western blot),实时荧光定量聚合酶链式反应(Real-time PCR)检测Bcl-2、Bax、Caspase-3蛋白与mRNA的表达情况,膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)/碘化丙锭(PI)法流式细胞术检测各组SCs的凋亡率。
结果
2
Western blot,Real-time PCR检测结果显示,与正常组比较,高糖组Bcl-2蛋白及mRNA表达水平降低,Bax、Caspase-3蛋白及mRNA表达水平升高,差异有统计学意义(
P
<
0.01);与高糖组比较,HPS+高糖组、
α
-LA+高糖组的Bcl-2蛋白及mRNA表达水平明显升高,Bax、Caspase-3蛋白及mRNA表达水平明显降低,差异有统计学意义(
P
<
0.01)。Annexin V/PI法流式细胞术检测结果显示,与正常组比较,高糖组SCs凋亡率明显增加;与高糖组比较,HPS+高糖组和
α
-LA+高糖组SCs凋亡率显著降低,差异有统计学意义(
P
<
0.01)。
结论
2
HPS可以减轻SCs凋亡反应状态,其作用机制可能与抑制Bcl-2/Caspase-3信号通路激活有关。
Objective
2
To observe the effects of Hedysarum polysaccharides(HPS)on the signaling pathways of B-cell lymphoma 2 (Bcl-2), cysteinyl aspartate-specific protease 3 (Caspase-3), and Bcl-2-associated X protein (Bax) in Schwann cells(SCs)cultured in high glucose,and explore the possible mechanism of HPS against diabetic peripheral neuropathy(DPN).
Method
2
Four SD suckling mice aged 5-7 days were randomly divided into a normal group,a high-glucose group,an HPS + high-glucose group,and an
α
-lipoic acid(
α
-LA)+ high-glucose group. SCs were extracted from the sciatic nerve and cultured in a 37 ℃,5% CO
2
incubator. After the cells reached 80% confluence,Cell Counting Kit-8(CCK-8)was used to screen the experimental concentrations suitable for high glucose,HPS, and α-LA interventions. Western blot and Real-time polymerase chain reaction (Real-time PCR)were used to detect the protein and mRNA expression of Bcl-2,Bax,and Caspase-3. The apoptosis rate of SCs was detected by flow cytometry using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI).
Result
2
As revealed by Western blot and real-time PCR,compared with the normal group,the high-glucose group showed reduced protein and mRNA expression of Bcl-2 and increased protein and mRNA expression of Bax and Caspase-3(
P
<
0.01). Compared with the high-glucose group,the HPS + high-glucose group and the
α
-LA + high-glucose group showed increased protein and mRNA expression of Bcl-2 and decreased protein and mRNA expression of Bax and Caspase-3(
P
<
0.01). As displayed by the results of flow cytometry using Annexin V/PI, compared with the normal group,the high-glucose group showed increased apoptosis rate;compared with the high-glucose group,the HPS + high-glucose group and the
α
-LA + high-glucose group showed reduced apoptosis rate(
P
<
0.01).
Conclusion
2
HPS can alleviate the apoptotic response of SCs,and its mechanism may be related to the inhibition of the activation of the Bcl-2/Caspase-3 signaling pathway.
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