Changes of auditory function and mechanisms in tinnitus rats treated by Wuling Capsule

doi:10.3969/j.issn.2095-4344.1951

Abstract

Abstract:

BACKGROUND: There is no standard cure for tinnitus. Wuling Capsule is a pure Chinese medicine preparation extracted from Wuling Ginseng by modern biological technology. Its composition is Wuling powder, which has the functions of nourishing the heart and calming the nerves, nourishing the kidney and strengthening the brain.
OBJECTIVE: To explore the effect of Wuling Capsule on the auditory function of tinnitus rats and its neuronal plasticity related genes in auditory central ultrastructure and auditory pathway.
METHODS: Seventy-five SPF-level Wistar rats were selected. In addition to the normal group, the other groups of rats were prepared with tinnitus model. After successful modeling, the model group (n=15) and the normal group (n=15) were given normal saline, and the carbamazepine group (n=15) was given carbamazepine solution. The low- and high-dose Wuling Capsule groups were intragastrically administered with 40 and 80 g of Wuling Capsule for 4 weeks. The changes of drinking water inhibition rate Rb, open field behavioral changes, auditory brainstem response threshold, primary auditory cortical pathology, auditory cortex ultrastructural changes, auditory central neurotransmitter γ-aminobutyric acid, 5-hydroxytryptamine were observed in the cortex, cochlear nucleus, inferior colliculus and upper olive.
RESULTS AND CONCLUSION: (1) The mean Rb in the high- and low-dose Wuling Capsule groups and the carbamazepine group in the conditioned reflex period (within 7 days) was significantly lower than that in the model group (P < 0.05). (2) On day 15 after modeling, the total stroke, average speed and erecting times in the model group and each drug-administered group were significantly lower than those in the normal control group, and the number of fecal pellets was significantly higher than that in the normal control group (P < 0.05). After the treatment, the total stroke, average speed and erecting times in the drug-administered groups were significantly higher than those in the model group, and the number of fecal pellets was significantly lower than that in the model group (P < 0.05). (3) Compared with the model group, the levels of γ-aminobutyric acid and 5-hydroxytryptamine in the hypothalamus were significantly increased (P < 0.05), and expression of cytoskeletal proteins in the auditory cortex, inferior colliculus and cochlear nucleus decreased in the low-and high-dose Wuling Capsule groups (P < 0.05). (4) These results indicate that Wuling Capsule significantly reduces the degree of neuronal damage in the auditory pathway of tinnitus rats, improves the ultrastructure of the auditory center, and lowers the auditory brainstem response threshold, thereby improving the auditory function of tinnitus rats and improving the anxiety and depression. The mechanism may be increasing the levels of γ-aminobutyric acid and 5-hydroxytryptamine in the hypothalamus and improving the expression of neuronal plasticity-related genes in the auditory pathway of tinnitus rats. The study was approved by the Ethical Committee of Liuzhou General Hospital, approval No. TRH58689.

Key words: tinnitus, auditory central ultrastructure, auditory pathway, neuronal plasticity related genes, Wuling Capsule, γ-aminobutyric acid, growth response gene-1, nerve growth related protein, cytoskeletal protein

CLC Number:

Mo Binyu, Wei Tao, Li Jihui, Feng Haiyan, Chen Meiqiu, Wei Yumei. Changes of auditory function and mechanisms in tinnitus rats treated by Wuling Capsule[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(35): 5645-5651.

Figures/Tables 7

2.1 动物数量分析大鼠75只，15只为正常组，实验期间均正常饮食水，无死亡；60只耳鸣模型组大鼠中，53只造模成功，其中，5只造模过程中死亡，2只顺应性较差造模不成功，造模成功率为88%。后期实验补充了模型大鼠7只，每组均为15只。 2.2 大鼠耳鸣情况乌灵胶囊低、高剂量组及卡马西平组大鼠在条件反射消减期(7 d内)的反射抑制率Rb的均值显著低于模型组(P < 0.05)。提示乌灵胶囊及卡马西平对水杨酸钠造成耳鸣的症状具有一定的改善作用，且乌灵胶囊组的Rb值恢复速度慢于卡马西平组，提示乌灵胶囊对耳鸣的改善作用优于卡马西平组，见图1。 2.3 各组大鼠旷场行为学变化实验前，各组大鼠的运动总行程、速度、直立次数、中央格停留时间及粪便数量比较差异均无显著性意义(P > 0.05)。造模后15 d，模型组及各给药组的运动总行程、平均速度和直立次数均显著少于正常组，粪便粒数显著多于正常组(P < 0.05)。治疗结束后，各给药组运动总行程、平均速度和直立次数均显著多于模型组，粪便粒数显著少于模型组(P < 0.05)，见表2。 2.4 不同声音刺激频率下各组大鼠ABR阈值情况和正常组相比，不同刺激频率下模型组大鼠ABR阈值上升；和模型组相比，不同刺激频率下卡马西平组及乌灵胶囊低、高剂量组大鼠ABR阈值显著降低(P < 0.05)，见表3。 2.5 各组大鼠大脑初级听皮质病理情况正常组大鼠神经元细胞紧密、规则排列；模型组大鼠神经元细胞分布不均匀，细胞膜为空泡状，毛细血管变形弯曲，部分细胞发生皱缩；乌灵胶囊低、高剂量组大鼠神经元细胞不同程度好转，且乌灵胶囊高剂量组大鼠改善效果最佳；卡马西平组大鼠神经元细胞受损改善情况不明显，见图2。 2.6 各组大鼠听皮质超微结构情况和正常组相比，模型组大鼠突触活性区长度、突触前囊泡、突触曲率、突触间隙及突触后致密区厚度增加；和模型组相比，乌灵胶囊低、高剂量组大鼠突触活性区长度、突触前囊泡、突触曲率、突触间隙及突触后致密区厚度显著降低(P < 0.05)，而卡马西平组听皮质突触参数改善不明显。详见表4，图3。 2.7 各组大鼠脑组织中γ-氨基丁酸及5-羟色胺的水平变化与正常组相比，模型组大鼠下丘脑γ-氨基丁酸及5-羟色胺水平显著降低(P < 0.05)。与模型组相比，乌灵胶囊低、高剂量组大鼠下丘脑γ-氨基丁酸及5-羟色胺水平显著增加(P < 0.05)，而卡马西平组大鼠下丘脑γ-氨基丁酸及5-羟色胺水平水平变化不明显(P > 0.05)，见表5。"

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