Study of the mechanism of Baizhu-Huangqi-Zhishi Formula in the treatment of constipation in Parkinson disease based on network pharmacology and its experimental verification
BAO Zhongming1,2 WEN Daiwei3 LU Pingang4 HUANG Wubiao1 TANG Guowen1
1.Department of Encephalopathy, Traditional Chinese Medicine Hospital of Wuzhou City, Guangxi Zhuang Autonomous Region, Wuzhou 543002, China; 2.Department of Neurology, the Sixth Affiliated Hospital of Guangxi University of Chinese Medicine, Guangxi Zhuang Autonomous Region, Wuzhou 543002, China;
3.Department of Spleen and Stomach, Traditional Chinese Medicine Hospital of Wuzhou City, Guangxi Zhuang Autonomous Region, Wuzhou 543002, China; 4.Department of Encephalopathy, Traditional Chinese Medicine Hospital of Wuzhou City, Guangxi Zhuang Autonomous Region, Wuzhou 543002, China
Abstract：Objective To explore the mechanism of Baizhu-Huangqi-Zhishi Formula in the treatment of constipation in Parkinson disease based on network pharmacology and to conduct its experimental verification. Methods The main active components of Rhizoma Atractylodis Macrocephalae, Radix Astragali seu Hedysari and Fructus Aurantii Immaturus were screened by network pharmacology, and the functional analysis and signal pathway analysis were carried out to analyze the molecular mechanism of the active compound-target network in treating constipation in Parkinson disease. Twenty healthy male SD rats aged six weeks were selected, and 15 of them were double injected with 6-hydroxydopamine to establish the model of Parkinson disease. After success, the constipation model of Parkinson disease was established by gavage with Loperamide 3 mg/kg. After successful modeling, the rats were divided into model group, positive control group and experimental group according to random number table method, with five rats in each group and other five rats as the normal group. The experimental group was given 10 ml/kg concentrated solution of Baizhu-Huangqi-Zhishi Formula by gavage. The positive control group was given Metobar 50 mg/kg and polyethylene glycol 1 mg/kg by gavage. The normal group and the model group were given the same amount of normal saline, once a day, for two weeks. Two weeks later, the gene and protein expressions of mitogen-activation protein kinase 8 (MAPK8), interleukin-6 (IL-6), MYC, and cysteine aspartic acid specific protease 3 (caspase 3) in colon of the four groups were detected by quantitative real-time PCR (qRT-PCR) and Western blot. Results Through database and literature screening, a total of 50 active molecules were obtained from Rhizoma Atractylodis Macrocephalae, Radix Astragali seu Hedysari and Fructus Aurantii Immaturus, and 116 action targets were identified, 85 of which were constipation in Parkinson disease. The main nodes of protein-protein interaction network were MAPK8, epidermal growth factor receptor, Cyclin D1, IL-6, MYC, and vascular endothelial growth factor A, etc. Functional analysis and signal pathway analysis showed that Baizhu-Huangqi-Zhishi Formula was involved in constipation in Parkinson disease mainly through biological processes such as ubiquitin-like protein ligase binding and ubiquitin protein ligase binding. At the same time, it may be possible to treat constipation in Parkinson disease through PI3K-Akt, MAPK, neurodegeneration-multiple diseases and other signaling pathways. The results of qRT-PCR and Western blot showed that the expression levels of MAPK8 gene and protein in the model group were lower than those in the normal group, and the expression levels of IL-6, MYC, caspase 3 gene and protein in the model group were higher than those in the normal group. The expression levels of MAPK8 gene and protein in the experimental group were higher than those in the model group, and the expression levels of IL-6, MYC, caspase 3 gene and protein in the experimental group were lower than that in the model group, with statistical significance (all P ＜ 0.05). Conclusion Baizhu-Huangqi-Zhishi Formula can achieve the therapeutic effect constipation in Parkinson disease well through multiple targets and multiple pathways.
包仲明1,2 文黛薇3 陆品刚4 黄舞标1 唐国文1. 基于网络药理学研究白术-黄芪-枳实组方治疗帕金森病便秘的作用机制及实验验证[J]. 中国医药导报, 2021, 18(19): 4-12.
BAO Zhongming1,2 WEN Daiwei3 LU Pingang4 HUANG Wubiao1 TANG Guowen1. Study of the mechanism of Baizhu-Huangqi-Zhishi Formula in the treatment of constipation in Parkinson disease based on network pharmacology and its experimental verification. 中国医药导报, 2021, 18(19): 4-12.
[1] Global Parkinson’s Disease Survey（GPDS）Steering Committee. Factors impacting on quality of life in Parkinson disease：results from an international survey [J]. Mov Disord，2002，17（1）：60-67.
[2] 韦晓芸，胡玉英，张诗敏，等.帕金森病所致便秘的中西医治疗进展[J].湖南中医杂志，2020，36（11）：201-203.
[3] 李悠悠，姜希娟，王阳雪，等.中医药治疗帕金森病伴功能性便秘的研究进展[J].中西医结合心脑血管病杂志，2020，18（5）：769-772.
[4] 贡钰霞，王浩，谷云飞.基于运脾理论探讨慢性功能性便秘的证治特点[J].南京中医药大学学报，2019，35（5）：567-569.
[5] 王晓晓，付文玉，庄文欣，等.改良纹状体内两点注射6-OHDA法制备大鼠帕金森病模型[J].神经解剖学杂志，2015，31（2）：187-192.
[6] 章洪鹏，徐家明，汤东，等.洛哌丁胺、硫糖铝和限水法制作便秘大鼠模型的效果对比研究[J].中国现代普通外科进展，2018，21（8）：594-598.
[7] 裴媛，马进，李姗，等.独活和独活香豆素对帕金森病模型大鼠脑黑质CHOP和Caspase12表达的影响[J].中华中医药学刊，2019，37（11）：2579-2582.
[8] More S，Choi DK. Neuroprotective Role of Atractylenolide-Ⅰ in an In Vitro and In Vivo Model of Parkinson’s Disease [J]. Nutrients，2017，9（5）：451.
[9] 何英杰，刘东波，唐其，等.酸橙类中药材枳实和枳壳化学成分研究进展[J].中药材，2017，40（6）：1488-1494.
[10] 张红，孙明江，王凌.枳实的化学成分及药理作用研究进展[J].中药材，2009，32（11）：1787-1790.
[11] 张蔷，高文远，满淑丽.黄芪中有效成分药理活性的研究进展[J].中国中药杂志，2012，37（21）：3203-3207.
[12] 张晓娟，左冬冬.白术化学成分及药理作用研究新进展[J].中医药信息，2018，35（6）：101-106.
[13] 李金容，陈倩，陶文强，等.二白汤治疗帕金森病相关便秘的临床疗效观察[J].河北中医，2017，39（9）：1337-1339.
[14] Liu J，Liu W，Lu Y，et al. Piperlongumine restores the balance of autophagy and apoptosis by increasing BCL2 phosphorylation in rotenone-induced Parkinson disease models [J]. Autophagy，2018，14（5）：845-861.
[15] Sliter DA，Martinez J，Hao L，et al. Parkin and PINK1 mitigate STING-induced inflammation [J]. Nature，2018， 561（7722）：258-262.
[16] 曾奇，刘智斌，王渊，等.帕金森病便秘研究进展[J].辽宁中医药大学学报，2020，22（8）：40-44.
[17] 李悠悠，姜希娟，王阳雪，等.中医药治疗帕金森病伴功能性便秘的研究进展[J].中西医结合心脑血管病杂志，2020，18（5）：769-772.
[18] 刘志强，王博龙.中药网络药理学药效成分筛选与靶标预测的研究进展[J].中成药，2019，41（1）：171-178.
[19] Chi J，Xie Q，Jia J，et al. Integrated Analysis and Identification of Novel Biomarkers in Parkinson’s Disease [J]. Front Aging Neurosci，2018，10：178.
[20] Herrán E，Requejo C，Ruiz-Ortega JA，et al. Increased antiparkinson efficacy of the combined administration of VEGF- and GDNF-loaded nanospheres in a partial lesion model of Parkinson’s disease [J]. Int J Nanomedicine，2014，9：2677-2687.
[21] O’Flanagan CH，Morais VA，Wurst W，et al. The Parkinson’s gene PINK1 regulates cell cycle progression and promotes cancer-associated phenotypes [J]. Oncogene，2015，34（11）：1363-1374.
[22] Yu MY，Yeo JH，Park JS，et al. Long-term efficacy of oral calcium polystyrene sulfonate for hyperkalemia in CKD patients [J]. PLoS One，2017，12（3）：e0173542.
[23] Wang R，Yang S，Nie T，et al. Transcription Factors：Potential Cell Death Markers in Parkinson’s Disease [J]. Neurosci Bull，2017，33（5）：552-560.
[24] Hampe C，Ardila-Osorio H，Fournier M，et al. Biochemical analysis of Parkinson’s disease-causing variants of Parkin，an E3 ubiquitin-protein ligase with monoubiquitylation capacity [J]. Hum Mol Genet，2006，15（13）：2059-2075.
[25] Blanca Ramírez M，Madero-Perez J，Rivero-Rios P，et al. LRRK2 and Parkinson’s Disease：From Lack of Structure to Gain of Function [J]. Curr Protein Pept Sci，2017，18（7）：677-686.
[26] Yan T，Sun Y，Gong G，et al. The neuroprotective effect of schisandrol A on 6-OHDA-induced PD mice may be related to PI3K/AKT and IKK/IκBα/NF-κB pathway [J]. Exp Gerontol，2019，128：110743.
[27] 曾蓉，喻斌，徐寅，等.灭幽汤对幽门螺杆菌相关性胃炎脾胃湿热证小鼠PTEN-PI3K-Akt-FoxO凋亡信号通路的影响[J].现代中西医结合杂志，2020，29（26）：2869-2875.
[28] Yang J，Jia M，Zhang X，et al. Calycosin attenuates MPTP-induced Parkinson’s disease by suppressing the activation of TLR/NF-κB and MAPK pathways [J]. Phytother Res，2019，33（2）：309-318.
[29] Singh S，Mishra A，Srivastava N，et al. MK-801（Dizocilpine）Regulates Multiple Steps of Adult Hippocampal Neurogenesis and Alters Psychological Symptoms via Wnt/β-Catenin Signaling in Parkinsonian Rats [J]. ACS Chem Neurosci，2017，8（3）：592-605.
[30] Choi HY，Mai TH，Kim KA，et al. Association between viral hepatitis infection and Parkinson’s disease：A population-based prospective study [J]. J Viral Hepat，2020， 27（11）：1171-1178.
[31] 唐莉莉.基于脑肠轴理论探讨温肾养肝方调节肠道菌群治疗帕金森病的临床观察和实验研究[D].南京：南京中医药大学，2020.
[32] West AB，Kapatos G，O’Farrell C，et al. N-myc regulates parkin expression [J]. J Biol Chem，2004，279（28）：28896-28902.
[33] 陈林，张勇.加味白术汤治疗功能性便秘80例疗效观察[J].浙江中医杂志，2020，55（8）：578-579.
[34] 姜亚欣.白术内酯Ⅰ对慢传输型便秘大鼠结肠水通道蛋白3、4表达的调节作用和机制[D].青岛：青岛大学，2017.
[35] 田雨，丁艳平，邵宝平，等.黄芪等药食同源类中药作为功能性食品与肠道菌群的相互作用[J].中国中药杂志，2020，45（11）：2486-2492.
[36] 谢绍锋，黄莉吉，狄红杰，等.中医药基于肠道菌群调节防治肥胖研究进展[J].世界科学技术-中医药现代化，2019，21（11）：2474-2479.
京公网安备 11010502046598号