阅读说明：本技术 春桑叶水提取物在制备治疗亨廷顿病药物中的应用 (Application of spring mulberry leaf water extract in preparation of medicine for treating Huntington disease ) 是由 支德娟 徐帅帅 崔伟薇 张响 汤杨婧 张瑞 付献瑶 于 2020-06-28 设计创作，主要内容包括：本发明属于医药领域,具体涉及春桑叶水提取物在制备治疗亨廷顿病药物中的应用。本发明发现初春采摘的春桑叶水提取物,能够显著抑制突变多聚谷氨酰胺蛋白(PolyQ蛋白)的聚集,这表明春桑叶水提取物显著减轻了亨廷顿病样病理特征,可被用于制备治疗亨廷顿病药物。(The invention belongs to the field of medicines, and particularly relates to an application of a spring mulberry leaf water extract in preparation of a medicine for treating Huntington disease. The invention discovers that the water extract of the mulberry leaves picked in early spring can obviously inhibit the aggregation of mutant polyglutamine protein (PolyQ protein), which indicates that the water extract of the mulberry leaves in spring obviously relieves the pathological characteristics of Huntington diseases and can be used for preparing the medicine for treating the Huntington diseases.)

1. Application of water extract of folium Mori in preparing medicine for treating Huntington disease is provided.

2. The use of claim 1, wherein the preparation method of the water extract of mulberry leaf in spring comprises: adding 7.5 volumes of boiling water into the spring mulberry leaves, soaking for 30min, and filtering by filter paper to obtain the water extract of the spring mulberry leaves.

3. The application of claim 1, wherein the preparation of the water extract of folium mori is any one of tablets, capsules, powder, syrup, granules, pills, mixture, dripping pills and pharmaceutically acceptable forms.

4. Application of a water extract of folium mori in preparing a medicine for inhibiting aggregation of mutant Poly Q protein.

5. The use of claim 4, wherein the water extract of mulberry leaf in spring is prepared by: adding 7.5 volumes of boiling water into the spring mulberry leaves, soaking for 30min, and filtering by filter paper to obtain the water extract of the spring mulberry leaves.

6. The use of claim 4, wherein the water extract of mulberry leaf in spring is in the form of any one of tablets, capsules, powders, syrups, granules, pills, mixtures, dripping pills and pharmaceutically acceptable forms.

Technical Field

The invention belongs to the field of medicines, and particularly relates to an application of a spring mulberry leaf water extract in preparation of a medicine for treating Huntington disease.

Background

Huntington's Disease (HD) is a fully-dominant genetic disease with a single autosomal mutation. The disease is a delayed neurodegenerative disease in which accumulation of mutant huntingtin protein (Htt) causes cytotoxicity, mainly damaging basal ganglia and cerebral cortex. Its clinical symptoms are complex and include three major categories, namely motor symptoms, cognitive dysfunction and mental disorders. Patients usually develop disease around 40 years of age, with worsening disease over time, and die after about 15-20 years (Zhushu et al, 2017; Jean-MarcBurgunder et al, 2015).

It is now well recognized that the development of HD is caused by the excessive expansion of the glutamine sequence repeated at the amino terminus of Htt, but the specific pathogenic mechanism of mutant Htt remains to be explored. The Htt gene is located on human chromosome IV and is responsible for encoding the Htt protein, and a polyglutamine (Poly Q) fragment encoded by the repeated codon CAG is present at the amino terminus. The normal repeat sequence is 6-35, and when the repeat sequence is abnormally larger than 36, HD will be caused. The insoluble protein resulting from aggregation of the N-terminal fragment of mutated Htt is found in the brain of patients, and toxicity affects the development of the nervous system of the brain and the function of nervous tissues. The mutant protein has aggregation forms of polymer and nuclear inclusion body, the tissue forms and histochemical properties of the mutant protein are the same, the mutant protein is only different from the occurrence part, and the amino-terminal Poly Q segment of the protein is the main component of the polymer. Mutant Htt interferes with nuclear gene expression, affects neural development and induces cell death, is associated with mitochondrial dysfunction, and toxicity occurs due to accumulation of protein resulting from clearance disorders (chenvirejie et al, 2019; qin zheng hong et al, 2004).

Currently, no effective treatment method for significantly preventing or reversing the course of disease exists for HD, and drugs are mainly developed for relieving clinical symptoms. Clinically, drugs for HD treatment include drugs for alleviating excitotoxicity, such as reserpine and amantadine; drugs that inhibit the activity of Caspases (a class of cysteine proteases that cleave mutated Htt to form more toxic Htt fragments), such as minocycline; aggregation-reducing agents, such as rapamycin; drugs that improve mitochondrial function, such as coenzyme Q10; drugs that regulate gene transcription abnormalities, such as suberoylanilide hydroxamic acid and the like (Seattle et al, 2011). These drugs can only improve various symptoms to a certain extent, but all have deficiencies, and the effective therapeutic drugs are still lacking clinically. Studies have shown that Poly Q fragment of Htt mutant protein is an important factor in pathogenesis, and therefore, the development of anti-huntington's disease should be initiated by reducing the expression of Poly Q protein.

The mulberry leaf is derived from dry leaves of Morus alba L, a traditional Chinese medicine in China, and is recorded in Shen nong Ben Cao Jing earliest. Modern pharmacological experimental studies prove that the mulberry leaves have various pharmacological effects of reducing blood sugar, reducing blood pressure, regulating blood fat, resisting inflammation, resisting virus, resisting oxidation, resisting aging and the like. In addition, mulberry leaves have the functions of tonifying and stabilizing the nervous system, promoting nerve cell growth factors, protecting nerve cells and preventing damage of nerve cells, and active ingredients in the mulberry leaves comprise flavone, flavone glycosides, alkaloids, polysaccharides, sterols, volatile oils, amino acids, vitamins and trace elements (euyanzhen et al, 2003). Generally, mulberry leaves are classified into spring mulberry, frost mulberry and mulberry leaves picked between them. At present, spring mulberries are mostly eaten, silkworms are bred after picking, and frost mulberries are collected after first frost and then put into medicines. The frosted mulberry is obtained by removing impurities after picking and drying in the sun, is a wind-heat dispersing medicine and can be taken orally or applied externally. It is sweet in taste, neutral in nature and cold in nature, has the effects of dispelling wind and heat, clearing lung-heat, moistening dryness, benefiting liver, ventilating, nourishing yin, replenishing blood and the like, and is a medicine and food dual-purpose article (Zhao Li jade and the like, 2004). The invention provides application of spring mulberry leaves in preparation of a medicine for treating Huntington diseases, and a large number of documents show that although the mulberry leaves are frosted to be frost mulberry which is the best to be used as the medicine, the spring mulberry leaves picked at the beginning of spring have obvious treatment effect in treatment of the Huntington diseases, which indicates that the spring mulberry leaves have the potential for treating the Huntington diseases and can be applied in preparation of the medicine for treating the Huntington diseases.

The Caenorhabditis elegans (Caenorhabditis elegans) is used as a model organism, and is a multifunctional medicine screening and medicine action mechanism research tool. It is cheap and easy to culture; the generation period is short, the number of offspring is large, a large number of individuals with consistent backgrounds can be obtained, the experimental repeatability is ensured, and the experiment is carried out by adopting a large sample amount, so that the influence of individual difference is eliminated; and higher organisms are highly conserved in gene and molecular signal pathways (zeia, et al, 2008), so that the method is widely applied to the molecular mechanism of neurodegenerative diseases and the initial screening research of related drugs. polyQ and Yellow Fluorescent Protein (YFP) are fused and expressed in muscle cells of the body wall of the nematode, highly repeated polyQ presents dense fluorescent bright spots on the body wall of the nematode, the HD nematode is treated by adopting a tested drug, and the change of the fluorescence aggregation degree of polyQ in the muscle cells of the body wall is observed to judge whether the tested drug has the effect of treating HD (Kaletta and Hengartner, 2006).

Reference to the literature

[1] Zhushu, Tang Tie shan DNA damage repair and Huntington's disease mechanism [ J ] biochemical and biophysical progress, 2017,44:727-736.

[2] Jean-MarcBurgunder, Voluoyang Huntington chorea teaching review [ J ] J. China J. Neuromental diseases 2015,41: 577-.

[3] Clinical research progress of early diagnosis and treatment of ChenvireJie, Fomin, Huntington's disease [ J ] Chongqing medical university school newspaper, 2019,44: 531-.

[4] The molecular pathological study progress of Qinzhong, Gu Sha Lun, Lin Fang Huntington chorea [ J ]. Chinese pharmacological Notification, 2004,4: 378-one 382.

[5] Yan ya Ping, Zhang Bao Rong, Huntington's disease pathogenesis and treatment progress [ J ]. J.2011, 11:30-35, China's journal of modern neurological disease.

[6] Zhao Liyu jade, Qifeng lan, Dian Xiao Hua and Chen Xiang, the nutritive and health-care function of mulberry leaf and comprehensive utilization of its (J) Chinese food and nutrients (2004, 2: 23-26).

[7] The research progress of chemical components and pharmacological actions of the euyang, Chenjun, mulberry leaves [ J ]. the university of Jiangsu bulletin (Nature science edition), 2003,6:39-44.

[8] Chua, Wen Xin, Huangjianhua, Shenyi, Epimedium, and Epimedium general flavone for delaying the aging of caenorhabditis elegans [ J ] an experimental study of the Chinese & Western medicine integration journal, 2008,28:522 + 525.

[9]Kaletta T,Hengartner MO.Finding function in novel targers:C.elegans as a model organism.Nature Reviews Drug Discovery,2006,5:387-399.

[10] Zhang Qiang, Sunwang, Zhang Jing, content determination of polysaccharide and total flavone in mulberry leaf [ J ]. Jilin agriculture, 2011,3:73-74.

Disclosure of Invention

The invention aims to provide application of a spring mulberry leaf water extract in preparation of a medicine for treating Huntington disease.

Further, the preparation method of the water extract of the mulberry leaves in spring comprises the following steps: adding 7.5 volumes of boiling water into the spring mulberry leaves, soaking for 30min, and filtering by filter paper to obtain the water extract of the spring mulberry leaves.

Further, the dosage form of the water extract of the mulberry leaf in spring is any one of tablets, capsules, powder, syrup, granules, medicinal granules, pills, mixture, dropping pills and the pharmaceutically achievable dosage form.

The invention aims to provide application of a water extract of mulberry leaves in spring in preparation of a medicine for inhibiting aggregation of mutant Poly Q protein.

Further, the preparation method of the water extract of the mulberry leaves in spring comprises the following steps: adding 7.5 volumes of boiling water into the spring mulberry leaves, soaking for 30min, and filtering by filter paper to obtain the water extract of the spring mulberry leaves.

Further, the dosage form of the water extract of the mulberry leaf in spring is any one of tablets, capsules, powder, syrup, granules, medicinal granules, pills, mixture, dropping pills and the pharmaceutically achievable dosage form.

The mulberry leaves adopted by the invention are the mulberry leaves picked in the early spring, and are provided by Nanchong good tea Co. Preparing a water extract of spring mulberry leaves: adding 30mL of boiling water into 4g of spring mulberry leaves, soaking for 30min, and filtering by using filter paper to obtain 10mL of water extract of the spring mulberry leaves. The mass preparation of the water extract of the mulberry leaves in spring can be amplified according to the proportion, and the water extract of the mulberry leaves in spring is obtained by using corresponding heating and filtering equipment, which can be understood by those skilled in the art. The total flavonoids of mulberry leaves are used as the marker components of the water extract of the mulberry leaves in spring, and the content of the total flavonoids of the mulberry leaves is determined by a sodium nitrite-aluminum nitrate determination method.

The caenorhabditis elegans is taken as a model organism, the Poly Q segment is fused with YFP in nematode muscle cells through a gene recombination technology to be expressed as a caenorhabditis elegans HD pathological model, dense fluorescent bright spots can be observed under a fluorescent microscope, and the HD nematode can be remarkably reduced by treating the HD nematode with a drug candidate with anti-HD activity.

The beneficial effects of the invention are that the water extract of the mulberry leaves in spring is adopted to treat the tested caenorhabditis elegans, and the results show that: the water extract of the mulberry leaves in spring has obvious therapeutic effect on caenorhabditis elegans suffering from Huntington disease, which is shown in the obvious inhibiting effect on fluorescence aggregation caused by excessive aggregation of Poly Q protein in muscles of HD caenorhabditis elegans, which indicates that the water extract of the mulberry leaves in spring has the potential of treating the Huntington disease and can be applied to the preparation of medicines for treating the Huntington disease.

Drawings

FIG. 1 effect of different aqueous extracts of mulberry leaves on fluorescence aggregation of HD C. Note: c is a sterile water blank control; p is positive control group, 200 μ M epigallocatechin gallate (EGCG); CS is a spring mulberry leaf water extract experimental group; SS is the water extract experimental group of frost mulberry leaf; SF is the water extract of mulberry powder experimental group.

Detailed Description

The technical solution of the present invention is further described in detail with reference to the following specific examples, but the scope of the present invention is not limited to the following.