滋肾泻青汤对ADHD模型大鼠前额叶多巴胺受体脱敏-复敏信号通路的影响

孙继超; 张碧霞; 李伟伟

doi:10.14148/j.issn.1672-0482.2022.1137

滋肾泻青汤对ADHD模型大鼠前额叶多巴胺受体脱敏-复敏信号通路的影响

doi: 10.14148/j.issn.1672-0482.2022.1137

广西中医药大学第一附属医院, 广西南宁 530023

基金项目:

国家自然科学基金青年科学基金项目 81804146

国家自然科学基金地区科学基金项目 82260953

广西自然科学基金青年基金项目 2017JJB140344y

广西自然科学基金面上项目 2020JJA140275

广西高校中青年教师基础能力提升项目 2018KY0281

广西中医药大学引进博士科研启动基金项目 2017BS046

详细信息

作者简介:
孙继超, 男, 主治医师, E-mail: sunjichao881177@163.com

通讯作者:
李伟伟, 男，主任中医师, 博士生导师, 主要从事中医儿科相关疾病的研究, E-mail: 13878161612@163.com

中图分类号: R285.5
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出版历程
- 收稿日期: 2022-04-25
- 网络出版日期: 2022-12-15
- 发布日期: 2022-12-10

Effects of Zishen Xieqing Tang on Dopamine Signaling Pathway of Desensitization/Resensitization in Attention Deficit Hyperactivity Disorder Rats' Prefrontal Cortex

The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530023, China

摘要

摘要: 目的探讨中药方滋肾泻青汤对注意缺陷多动障碍模型大鼠(Spontaneously hypertensive rat, SHR)行为学表现及前额叶多巴胺受体脱敏-复敏信号通路的影响。方法 50只SHR大鼠随机分为模型组、利他林组(2 mg·kg^-1)、滋肾泻青汤低剂量组、中剂量组、高剂量组(6.0、12.1、24.1 g·kg^-1), 每组10只, 另设Wistar京都大鼠(Wistar Kyoto rat, WKY)为正常对照组。每日灌胃2次, 在此期间进行相应的行为学检测, 治疗4周后取大鼠前额叶组织。分别用Western blot及qPCR检测各组大鼠前额叶组织中GRK-6、β-arrestin2、NCS-1、PP2A及PSD-95等蛋白和mRNA表达水平。结果利他林及滋肾泻青汤能改善SHR大鼠的行为学表现。与正常组相比, 模型组大鼠前额叶中β-arrestin2、NCS-1、PP2A、PSD-95蛋白和mRNA表达水平有下调趋势或显著下调(P < 0.05), 而GRK-6的表达水平上调(P < 0.05);经治疗后, 与模型组相比, 利他林组、滋肾泻青汤各剂量组前额叶中β-arrestin2、NCS-1、PSD-95、PP2A蛋白和mRNA表达水平有上调趋势或明显上调(P < 0.05), 而GRK-6的表达水平明显降低(P < 0.05)。结论利他林及滋肾泻青汤能增加模型大鼠前额叶多巴胺受体的活性, 多巴胺受体脱敏-复敏信号通路可能是滋肾泻青汤治疗ADHD的潜在途径之一。
- 滋肾泻青汤 /
- 注意缺陷多动障碍 /
- 多巴胺受体 /
- 脱敏-复敏 /
- 中医药
Abstract: OBJECTIVE To investigate the effects of Zishen Xieqing Tang(ZXT)on attention deficit hyperactivity disorder (ADHD) related behavior and dopamine signaling pathway of desensitization and resensitization in spontaneously hypertensive rats' prefrontal cortex. METHODS 10 Wistar Kyoto rats were set as normal control group. 50 SHR rats were randomly divided into 5 groups including model group, methylphenidate (MPH) group (2 mg·kg^-1by gavage), and high, middle, low dose of ZXT groups(ig ZXT with the crude drug dosage 6.0, 12.1, 24.1 g·kg^-1 respectively). Rats were orally given the drugs twice a day, then, behavior experiment was done to test the ADHD related behavior changes. After 4 weeks of treatment, the prefrontal cortex of rats were collected for further study. Real-time PCR and Western blot were used to test the protein or mRNA expression of GRK-6, β-arrestin2, NCS-1, PP2A and PSD-95 in SHR rats' prefrontal cortex. RESULTS MPH group and ZXT groups could improve the ADHD related behavioral performance of SHR. Statistic difference of β-arrestin2, GRK-6, NCS-1, PSD-95, PP2A and PKA expression were observed among different groups (P < 0.05). Comparing with normal control group, the β-arrestin2, NCS-1, PSD-95 and PP2A expression of SHR rats' prefrontal cortex in model group were much lower (P < 0.05). After treatment with ZXT, comparing the model group, the expression of β-arrestin2, NCS-1, PSD-95 and PP2A in MPH group and every-dosage group of ZXT were much higher in both parts (P < 0.05). At the meanwhile, the expression of GRK-6 was observed on the contrary behavior. CONCLUSION Both MPH and Zishen Xieqing Tang can affect the function of dopamine receptors by regulating desensitization and re-sensitization, thus improve the ADHD related symptom.
- Zishen Xieqing Tang /
- attention deficit hyperactivity disorder /
- dopamine receptor /
- desensitization-resensitization /
- Chinese medicine

HTML全文

图 1 滋肾泻青汤方对各组大鼠前额叶多巴胺受体信号通路中β-arrestin2、PSD-95、NCS-1、GRK-6、PP2A蛋白表达的影响

注: WKY+DDW.正常对照组; SHR+DDW.模型组; SHR+MPH. 利他林组; SHR+ZXT-L、M、H.滋肾泻青汤低、中、高剂量组。与WKY+DDW组比较, ^*P < 0.05;与SHR+DDW组比较, ^#P < 0.05。x±s, n=6。

Figure 1. The regulatory effects of Zishen Xieqing Tang on dopamine receptor signaling pathway related proteinβ-arrestin2, PSD-95, NCS-1, GRK-6, PP2A in rats' prefrontal cortex

下载: 全尺寸图片幻灯片

图 2 滋肾泻青汤方对各组大鼠前额叶多巴胺受体信号通路中β-arrestin2、PSD-95、NCS-1、GRK-6、PP2A mRNA表达的影响

注: WKY+DDW.正常对照组; SHR+DDW.模型组; SHR+MPH.利他林组; SHR+ZXT-L、M、H.滋肾泻青汤低、中、高剂量组。与正常组比较, ^*P < 0.05;与模型组比较, ^#P < 0.05。x±s=6。

Figure 2. The regulatory effects of Zishen Xieqing Tang on dopamine receptor signaling pathway related mRNA expression in rats' prefrontal cortex

下载: 全尺寸图片幻灯片

表 1 引物序列

Table 1. Primer sequences

基因	引物序列(5'→3')	产物长度/bp
GAPDH	Forward: GACATGCCGCCTGGAGAAAC	92
	Reverse: AGCCCAGGATGCCCTTTAGT
GRK-6	Forward: CAGTCCAGATGAGCAAGCAG	103
	Reverse: CACAACAGACAGGCACGAGT
β-arrestin2	Forward: AGGGCAGTGGGATACAGGT	108
	Reverse: AGGGCAAACAAAGCAAACAG
NCS-1	Forward: GCTGTGATGTGTGGGAACC	101
	Reverse: GCTTTGTGGAGACGAGGAAG
PSD-95	Forward: CACTGACAACCCGCACATC	136
	Reverse: CTCCCGAACATCCACTTCAT
PP2A	Forward: TTACCGAGAGCGTATCACCA	171
	Reverse: ATCTGCCCATCCACCAAG

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表 2 滋肾泻青汤对SHR大鼠开场实验活动距离的影响(x±s, m, n=10)

Table 2. The effects of Zishen Xieqing Tang on the SHR rats' moving distances in open field test (x±s, m, n=10)

组别	给药前	给药2周	给药4周
正常组	11.14±1.83	13.21±2.02	11.96±1.68
模型组	31.11±3.31^**	33.81±2.63^**	33.14±3.57^**
利他林组	30.68±3.11^**	28.32±3.49^**#	23.09±2.63^**##
滋肾泻青汤低剂量组	29.59±3.73^**	27.64±2.51^**#	24.48±1.73^**##
滋肾泻青汤中剂量组	33.58±1.89^**	24.37±1.58^**##	25.63±1.73^**##
滋肾泻青汤高剂量组	31.74±3.26^**	26.93±2.30^**##	24.38±2.15^**##
注: 与正常组比较, ^{* *}P < 0.01;与模型组比较, ^#P < 0.05, ^##P < 0.01。

下载: 导出CSV

表 3 滋肾泻青汤对SHR大鼠水迷宫隐蔽站台实验潜伏期的影响(x±s, s, n=10)

Table 3. The effects of Zishen Xieqing Tang on the SHR rats' escape latency in Morris water maze test(x±s, s, n=10)

组别	第1天	第2天	第3天	第4天	第5天
正常组	81.22±7.51	83.72±7.25	79.56±9.32	81.24±6.96	83.65±8.14
模型组	70.33±6.57^**	66.35±8.24^**	55.32±4.97^**	46.84±3.75^**	32.05±5.63^**
利他林组	71.24±7.25^**	62.35±4.56^**	50.36±5.83^**	39.52±4.35^**#	20.31±3.52^**##
滋肾泻青汤低剂量组	73.64±6.35^**	67.56±7.52^**	51.36±6.21^**	33.65±5.46^**##	25.52±3.24^**##
滋肾泻青汤中剂量组	69.65±6.53^**	60.67±5.54^**#	53.25±3.54^**	41.36±4.13^**	23.25±4.68^**##
滋肾泻青汤高剂量组	72.34±8.63^**	62.36±5.63^**	49.36±4.25^**#	36.65±5.72^**##	21.35±4.61^**##
注: 与正常组比较, ^{* *}P < 0.01;与模型组比较, ^#P < 0.05, ^##P < 0.01。

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