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四生汤对电磁辐射所致大鼠海马神经元损伤的抑制作用
秦浩然1,骆华杰2,黄帅3,周西彬4,郑国银4,王丽娜5,顾伟6,黄念7*,凌昌全1*
0
(1. 海军军医大学(第二军医大学)第一附属医院中医肿瘤科, 上海 200433;
2. 上海交通大学医学院附属仁济医院耳鼻喉科, 上海 200001;
3. 上海市浦东医院骨科, 上海 201399;
4. 海军军医大学(第二军医大学)中医系经典著作教研室, 上海 200433;
5. 海军军医大学(第二军医大学)中医系中医内科教研室, 上海 200433;
6. 海军军医大学(第二军医大学)中医系军事中医学教研室, 上海 200433;
7. 海军军医大学(第二军医大学)第三附属医院中西医结合科, 上海 201805
*通信作者)
摘要:
目的 评价四生汤抑制电磁辐射所致大鼠海马神经元损伤的作用。方法 利用电磁辐射源辐射大鼠4周,低、中、高辐射功率密度分别为10、20及40 mW/cm2,构建大鼠电磁辐射损伤模型。以生理盐水为空白对照,安多霖胶囊为阳性对照,通过Morris水迷宫实验评估四生汤对大鼠因电磁辐射所致的学习记忆力下降的影响。利用TUNEL试剂盒和流式细胞术检测大鼠海马神经元的凋亡水平,采用细胞活性氧(ROS)试剂盒和流式细胞术检测大鼠海马神经元中ROS含量。结果 Morris水迷宫实验结果表明,高剂量辐射结束后第7天,空白对照组大鼠的平均逃避潜伏期(AEL)比四生汤组、安多霖组延长,安多霖组大鼠的AEL比四生汤组延长(均P<0.05)。中剂量辐射结束后第3天,空白对照组大鼠的AEL比四生汤组、安多霖组延长(均P<0.05);第14、28天时,空白对照组和安多霖组大鼠的AEL比四生汤组延长(均P<0.05)。低剂量辐射结束后,空白对照组大鼠的AEL则与四生汤组、安多霖组差异均无统计学意义(均P>0.05)。细胞凋亡检测结果证实,高剂量和中剂量比低剂量辐射的大鼠海马神经元凋亡率增大(均P<0.05)。高、中、低剂量辐射后,安多霖组和四生汤组大鼠的神经元凋亡率均低于空白对照组(均P<0.05)。高剂量辐射后,四生汤组大鼠的神经元凋亡率低于安多霖组(P<0.05)。ROS含量检测结果表明,高、中、低剂量辐射后大鼠的海马神经元ROS含量差异无统计学意义(P>0.05),但安多霖组和四生汤组大鼠海马神经元ROS含量均比空白对照组降低(均P<0.05)。中剂量辐射后,四生汤对大鼠海马神经元ROS的抑制作用比安多霖增强(P<0.05)。结论 四生汤能抑制电磁辐射导致的大鼠海马神经元凋亡、降低其细胞中ROS含量,从而减轻电磁辐射对神经元的损伤,缓解大鼠学习记忆力的下降。
关键词:  电磁辐射  四生汤  海马  神经损伤
DOI:10.16781/j.CN31-2187/R.20220592
投稿时间:2022-07-15修订日期:2022-08-09
基金项目:军事训练(单个人员训练)创新能力培养项目(2020SZ06-2).
Inhibitory effect of Sisheng decoction on hippocampal neuron injury induced by electromagnetic radiation in rats
QIN Haoran1,LUO Huajie2,HUANG Shuai3,ZHOU Xibin4,ZHENG Guoyin4,WANG Lina5,GU Wei6,HUANG Nian7*,LING Changquan1*
(1. Department of Traditional Chinese Medicine Oncology, The First Affiliated Hospital of Naval Medical University (Second Military Medical University), Shanghai 200433, China;
2. Department of Otolaryngology, Renji Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200001, China;
3. Department of Orthopaedics, Pudong Hospital, Shanghai 201399, China;
4. Department of Classical Work Office, School of Traditional Chinese Medicine, Naval Medical University (Second Military Medical University), Shanghai 200433, China;
5. Department of Internal Traditional Chinese Medicine, School of Traditional Chinese Medicine, Naval Medical University (Second Military Medical University), Shanghai 200433, China;
6. Department of Military Traditional Chinese Medicine, School of Traditional Chinese Medicine, Naval Medical University (Second Military Medical University), Shanghai 200433, China;
7. Department of Integrative Medicine, The Third Affiliated Hospital of Naval Medical University (Second Military Medical University), Shanghai 201805, China
*Corresponding authors)
Abstract:
Objective To evaluate the inhibitory effect of Sisheng decoction on the hippocampal neuron injury induced by electromagnetic radiation in rats. Methods Rats were irradiated with electromagnetic radiation sources for 4 weeks, with low-, medium-, and high-dose radiation power densities of 10, 20, and 40 mW/cm2, respectively. The Morris water maze was used to evaluate the effect of Sisheng decoction on the learning and memory decline caused by electromagnetic radiation in rats, with physiological saline as the blank control and anduolin capsule as the positive control. TUNEL kit and flow cytometry were used to detect the level of apoptosis in hippocampal neurons of rats with electromagnetic radiation injury, and reactive oxygen species (ROS) kit and flow cytometry were used to detect the ROS content in hippocampal neurons of rats. Results The results of the Morris water maze test showed that on the 7th day after high-dose radiation, the average escape latency (AEL) of the rats in the blank control group was significantly longer than that in the Sisheng decoction group and anduolin group, and the AEL of the rats in the anduolin group was significantly longer than that in the Sisheng decoction group (all P<0.05). On the 3rd day after medium-dose radiation, the AEL of the rats in the blank control group was significantly longer than that in the Sisheng decoction group and anduolin group (both P<0.05); on the 14th and 28th days, the AEL of the rats in the blank control group and anduolin group was significantly longer than that in the Sisheng decoction group (all P<0.05). After exposed to low-dose radiation, there was no significant difference in AEL between the blank control group and the Sisheng decoction or anduolin groups (both P>0.05). The apoptosis test result confirmed that the apoptosis rates of hippocampal neurons in rats exposed to high- and medium-dose radiation were significantly higher than that exposed to low-dose radiation (both P<0.05). After exposed to high-, medium-, and low-dose radiation, the neuronal apoptosis rates of the rats in the anduolin group and Sisheng decoction group were significantly lower than those in the blank control group (all P<0.05). After exposed to high-dose radiation, the apoptosis rate of neurons of the rats in the Sisheng decoction group was significantly lower than that in the anduolin group (P<0.05). There were no significant differences in ROS contents in hippocampal neurons among rats under high-, medium-, or low-dose radiation (P>0.05). However, the ROS contents in neurons of the anduolin group and Sisheng decoction group were significantly lower than that in the blank control group (both P<0.05). After exposed to medium-dose radiation, the ROS inhibition effect of the Sisheng decoction group was stronger than that in the anduolin group (P<0.05). Conclusion Sisheng decoction can inhibit the apoptosis of hippocampal neurons in rats, reduce the ROS content in their cells, and alleviate the injury to learning and memory caused by electromagnetic radiation in rats.
Key words:  electromagnetic radiation  Sisheng decoction  hippocampus  nerve injury