TW201900195A - Herbal composition for preventing or alleviating ischemic stroke - Google Patents
Herbal composition for preventing or alleviating ischemic stroke Download PDFInfo
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本發明提供一種中草藥組合物,其包含川芎,白芷,雞血藤,木賊,川紅花,綠茶或其組合;本發明更提供製備該種中草藥組合物以用於預防或改善血栓或血栓相關疾病之用途,以及該種中草藥組合物之製備方法。 The present invention provides a Chinese herbal medicine composition comprising Chuanxiong, Angelica, Spatholobus, Equisetum, Saffron, Green Tea or a combination thereof; the present invention further provides for preparing the Chinese herbal composition for preventing or ameliorating thrombosis or thrombosis-related diseases. Use, and a preparation method of the Chinese herbal medicine composition.
腦中風也稱作急性腦血管病變,為台灣僅次於癌症的第二大死因,若將癌症依照不同的種類來排名統計,則腦中風為所有單一類疾病致死率的第一位。腦中風屬血管性疾病,當腦血管被血液栓塊塞住而造成血液無法正常供應氧氣及養分至腦部時,就會發生腦中風的現象;一旦腦部發生血液供應不足的情形,在短短數秒內,腦部的細胞就會開始死亡。腦中風的種類主要分為缺血性和出血性兩大類。根據統計,有百分之八十的中風病患屬缺血性腦中風,出血性腦中風約佔百分之二十。 Stroke is also called acute cerebrovascular disease, which is the second leading cause of death in Taiwan after cancer. If cancer is ranked according to different types, stroke is the first cause of death for all single diseases. Brain stroke is a vascular disease. When the blood vessels of the brain are blocked by blood plugs and the blood cannot supply oxygen and nutrients to the brain normally, a stroke will occur. Once the blood supply to the brain is insufficient, it is short. Within a few seconds, the cells in the brain begin to die. The types of stroke are mainly divided into two categories: ischemic and hemorrhagic. According to statistics, 80% of stroke patients have ischemic stroke, and hemorrhagic stroke accounts for about 20%.
川芎對於治療頭痛有一定的效果。《神農本草經》曰:「主中風入腦頭痛,寒痹」。金元四大家之李東元亦云川芎可:「補血治血虛頭痛」。然而川芎辛燥,只適合用於偏寒者。 Chuanxiong has a certain effect on the treatment of headache. "Shen Nong's Herbal Classic" said: "The main stroke has a headache, cold and cold." Li Dongyuan, a member of the Jinyuan Four, is also a member of Yunchuan. However, Chuanxiong is dry and only suitable for people with partial cold.
川芎與白芷相配治病歷史久遠。《本草經集注》已指出「白芷為之(川芎)使」。白芷與川芎同為祛風止痛藥,白芷並可除濕,與川芎共用可加強川芎的藥效。 Chuanxiong and white peony have a long history of treatment. "Materia Medica" has pointed out that "white 芷 is the (Chuan-chuan)." Both Baiqi and Chuanxiong are hurricane painkillers, white peony and dehumidification, and sharing with Chuanxiong can enhance the efficacy of Chuanxiong.
除非另外特別加以定義,於此敘述中所用的名詞將具有熟知此技藝者可瞭解之普通且常見之含意。如整個申請案中所使用,下列的術語具有下面所述之含意:術語「個體」意指一動物。較佳地,該動物係一哺乳動物,例如小鼠、大鼠、人、狗、貓等等。在一較佳實施例中,該個體係人類。 Unless otherwise specifically defined, the terms used in this description will have the ordinary and common meanings that are known to those skilled in the art. As used throughout the application, the following terms have the meanings set forth below: The term "individual" means an animal. Preferably, the animal is a mammal, such as a mouse, a rat, a human, a dog, a cat, and the like. In a preferred embodiment, the system is human.
本發明將川芎與白芷先行炮製,再加上活血組合物等中草藥組合以組成全新配方,以腦部為主要目標,並以動物實驗評估其預防缺血性腦中風之療效。 The invention combines Chuanxiong and Baiqi, and adds a combination of Chinese herbal medicines such as activating blood composition to form a new formula, taking the brain as the main target, and evaluating the curative effect of preventing ischemic stroke by animal experiments.
本發明之目的係提供一種中草藥組合物,其中包含以乾材料重量計之川芎約0.5~1.5重量分,白芷約0.5~1.5重量分,雞血藤約1.5~4.5重量分,木賊約0.5~1.5重量分,川紅花約0.2~1重量分,綠茶約0.5~1.5重量分或其組合。較佳地,該中草藥組合物包含以乾材料重量計之川芎約1重量分,白芷約1重量分,雞血藤約3重量分,木賊約1重量分,川紅花約0.5重量分,以及綠茶約1重量分。 The object of the present invention is to provide a Chinese herbal medicine composition comprising about 0.5 to 1.5 parts by weight of the dry weight of the material, about 0.5 to 1.5 parts by weight of the white peony, about 1.5 to 4.5 parts by weight of the spatholobus, and about 0.5 to 1.5 for the thief. The weight is about 0.2 to 1 part by weight of the safflower, and about 0.5 to 1.5 parts by weight of the green tea or a combination thereof. Preferably, the Chinese herbal medicine composition comprises about 1 part by weight of Rhizoma Chuanxiong, about 1 part by weight of white peony, about 3 parts by weight of spatholobus, about 1 part by weight of squid, about 0.5 weight of safflower, and green tea. About 1 part by weight.
於本發明一具體實施例中,該中草藥組合物係用於預防或改善血栓形成或血栓形成相關疾病。於另一具體實施例中,該血栓形成相關疾病係指心血管疾病或腦血管疾病。於一更具體實施例中,該腦血管疾病係指缺血性腦中風。 In a specific embodiment of the invention, the herbal composition is used to prevent or ameliorate thrombosis or thrombosis related diseases. In another specific embodiment, the thrombosis-related disease refers to a cardiovascular disease or a cerebrovascular disease. In a more specific embodiment, the cerebrovascular disease refers to an ischemic stroke.
本發明另提供一種製備中草藥組合物以預防或改善血栓形成或血栓形成相關疾病之用途,其中該中草藥組合物包含以乾材料重量計之川芎約0.5~1.5重量分,白芷約0.5~1.5重量分,雞血藤約1.5~4.5重量分,木賊約 0.5~1.5重量分,川紅花約0.2~1重量分,綠茶約0.5~1.5重量分或其組合。較佳地,該中草藥組合物包含以乾材料重量計之川芎約1重量分,白芷約1重量分,雞血藤約3重量分,木賊約1重量分,川紅花約0.5重量分,綠茶約1重量分。 The present invention further provides a use for preparing a Chinese herbal medicine composition for preventing or ameliorating a thrombosis or a thrombosis-related disease, wherein the Chinese herbal medicine composition comprises about 0.5 to 1.5 parts by weight of the sorghum, and about 0.5 to 1.5 parts by weight of the white peony. The spatholobus is about 1.5~4.5 weight points, the wood thief is about 0.5~1.5 weight points, the safflower is about 0.2~1 weight, the green tea is about 0.5~1.5 weight points or a combination thereof. Preferably, the Chinese herbal medicine composition comprises about 1 part by weight of Chuanxiong, about 1 part by weight of white peony, about 3 parts by weight of spatholobus, about 1 part by weight of squid, about 0.5 weight of safflower, and about about 50 parts by weight of green tea. 1 weight point.
本發明更提供一種一種製備中草藥組合物之方法,該方法包含:將0.5~1.5重量分川芎與0.5~1.5重量分白芷一起蒸煮半小時而成第一鍋煮液;將1.5~4.5重量分雞血藤與0.5~1.5重量分木賊煎煮二小時而成第二鍋煮液;將0.2~1重量分川紅花與0.5~1.5重量分綠茶加熱半小時而成第三鍋煮液;以及將第一鍋,第二鍋,以及第三鍋煮液混合而成一中草藥組合物之溶液。 The invention further provides a method for preparing a Chinese herbal medicine composition, which comprises: cooking 0.5~1.5 weight of Chuanxiong with 0.5~1.5 parts by weight of white peony for half an hour to form a first pot of cooking liquor; 1.5~4.5 weighting of spatholobus sinensis The second pot of cooking liquid is boiled with 0.5~1.5 weight of wood thief for two hours; the second pot cooking liquid is heated by adding 0.2~1 weight of safflower and 0.5~1.5 weight green tea for half an hour; and the first pot The second pot and the third pot of cooking liquid are mixed to form a solution of the Chinese herbal medicine composition.
於一具體實施例中,其中該中草藥組合物之溶液進一步加以濃縮而成一中草藥組合物之濃縮液。較佳地,其中該中草藥組合物之濃縮液進一步加以噴霧乾燥而成一中草藥組合物之粉末。 In one embodiment, the solution of the Chinese herbal composition is further concentrated to form a concentrate of a Chinese herbal composition. Preferably, the concentrate of the Chinese herbal medicine composition is further spray-dried to form a powder of a Chinese herbal medicine composition.
將本發明之中草藥組合物以腦動脈血管阻塞/再灌流中風模式小鼠進行試驗,腦中風試驗結果顯示本發明之中草藥組合物可有效預防缺血性腦中風所帶來的腦部傷害(圖一),以及改善中風所帶來之神經缺陷行為(圖二)。 The Chinese herbal medicine composition of the present invention was tested in a cerebral artery occlusion/reperfusion stroke mode mouse, and the results of the stroke test showed that the herbal composition of the present invention can effectively prevent brain damage caused by ischemic stroke. a), as well as improving the neurological deficits caused by stroke (Figure 2).
本發明的組合物可以以固體,溶液,乳劑,分散體,微膠粒,脂質體,以及其他如含有本發明中的一種或多種成分作為活性成分的組合物產物,或與有機或無機載體或賦形劑混合以適用於腸內或腸胃外的施用。活性成分可以被混合,例如,藥學上可接受的通常無毒性的載具如片劑,丸劑,膠囊,栓劑,溶液,乳液,懸浮液以及其他任何合適的形式以供使用。可使用的載體包括葡萄糖,乳糖,阿拉伯樹膠,明膠,甘露醇,澱粉糊,三矽酸 鎂,滑石,玉米澱粉,角蛋白,膠體二氧化矽,馬鈴薯澱粉,尿素,中等鏈長的甘油三酯,葡聚醣,以及合適用於製備製劑,固體,半固體或液體形式的其他載體。另外,亦可以使用穩定劑,增稠劑和著色劑和香料作為輔助。 The composition of the present invention may be in the form of a solid, a solution, an emulsion, a dispersion, a micelle, a liposome, and other composition products containing, as an active ingredient, one or more of the ingredients of the present invention, or with an organic or inorganic carrier or The excipients are mixed for application to enteral or parenteral administration. The active ingredient may be mixed, for example, in a pharmaceutically acceptable usually non-toxic vehicle such as a tablet, pill, capsule, suppository, solution, emulsion, suspension, and any other suitable form for use. Carriers which may be used include glucose, lactose, gum arabic, gelatin, mannitol, starch paste, magnesium tricaprate, talc, corn starch, keratin, colloidal cerium oxide, potato starch, urea, medium chain length triglycerides. , dextran, and other carriers suitable for the preparation of the formulation, solid, semi-solid or liquid form. In addition, stabilizers, thickeners and colorants and perfumes may also be used as an aid.
本發明之組合物可以口服的形式,例如作為片劑,錠劑,錠劑,水性或油性懸浮液,可分散粉末或顆粒,乳劑,硬或軟膠囊,或糖漿或酏劑。口服使用的組合物可根據各種已知的醫藥組合物製備方法加以製備,而此組合物可含有一種或多種如蔗糖,乳糖或糖精等甜味劑,如薄荷,冬青油或櫻桃等調味劑,著色劑和防腐劑以提供製藥上的美觀和口感。混合有活性成分與藥學上可接受無毒賦形劑的片劑也可藉由已知方法加以製造。可使用的賦形劑如:(1)惰性稀釋劑,如碳酸鈣,乳糖,磷酸鈣或磷酸鈉;(2)成粒劑和崩解劑,如玉米澱粉,馬鈴薯澱粉或海藻酸;(3)粘合劑,如黃蓍膠,玉米澱粉,明膠或阿拉伯膠,和(4)潤滑劑,如硬脂酸鎂,硬脂酸或滑石。片劑可為未包衣,或可以藉由已知技術進行包衣以延遲胃腸道中的崩解與吸收,從而提供較長時間的持續作用。例如延時材料如單硬脂酸甘油酯或二硬脂酸甘油酯均可採用,亦可藉由如美國專利案號4256108;4160452;及4,265,874所描述的技術包衣,以生成控制藥效釋放的滲透性治療片劑。 The compositions of the present invention may be administered orally, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions for oral use can be prepared according to various known pharmaceutical composition preparation methods, and the compositions may contain one or more sweeteners such as sucrose, lactose or saccharin, such as mint, wintergreen or cherry. Colorants and preservatives provide pharmaceutical aesthetics and mouthfeel. Tablets mixed with the active ingredient and a pharmaceutically acceptable non-toxic excipient can also be produced by known methods. Excipients which may be used are, for example, (1) inert diluents such as calcium carbonate, lactose, calcium phosphate or sodium phosphate; (2) granulating agents and disintegrating agents such as corn starch, potato starch or alginic acid; Adhesives such as tragacanth, corn starch, gelatin or gum arabic, and (4) lubricants such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period of time. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed, or may be coated by techniques such as those described in U.S. Patent Nos. 4,256,108; 4,160,452; and 4,265,874, to produce controlled release. Osmotic treatment tablets.
在某些情況下,口服使用的組合物可為硬明膠膠囊的形式,其中活性成分與惰性固體稀釋劑混合,例如碳酸鈣,磷酸鈣或高嶺土。它們也可為軟明膠膠囊的形式,其中活性成分與水或油介質混合,例如花生油,液體石蠟或橄欖油。 In some cases, the compositions for oral use can be in the form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin. They may also be in the form of soft gelatin capsules in which the active ingredient is mixed with water or oil medium, such as peanut oil, liquid paraffin or olive oil.
本發明之組合物實施例亦可為無菌注射用懸浮液的形式。此懸浮液可根據已知的方法使用適合的分散或濕潤劑和懸浮劑來配製。無菌注射用製劑亦可為無菌可注射溶液或懸浮液溶於無毒的腸道外可接受稀釋劑或溶劑中,例如作為在1,3-丁二醇中的溶液。無菌的不揮發油通常用作溶劑或懸浮介質。為了這個目的,任何溫和的固定油都可以使用,包括合成的單或二甘油酯,脂肪酸(包括油酸),天然存在的植物油如芝麻油,椰子油,花生油,棉籽油等,或合成的脂肪酸載體如油酸乙酯或類似物。緩衝劑,防腐劑,抗氧化劑等可以根據需要進行結合。 Embodiments of the compositions of the invention may also be in the form of a sterile injectable suspension. This suspension may be formulated according to known methods using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Sterile, fixed oils are usually employed as a solvent or suspension medium. For this purpose, any mild fixed oil can be used, including synthetic mono- or diglycerides, fatty acids (including oleic acid), naturally occurring vegetable oils such as sesame oil, coconut oil, peanut oil, cottonseed oil, etc., or synthetic fatty acid carriers. Such as ethyl oleate or the like. Buffering agents, preservatives, antioxidants, and the like can be combined as needed.
本發明之組合物實施例亦可以栓劑的形式於直腸腔投藥。透過將藥物與合適的非刺激性賦形劑,例如可可脂或聚乙二醇之合成甘油酯,混合而成組合物,此組合物於常溫下為固體,但在直腸腔液化和/或溶解以釋放藥物。 Embodiments of the compositions of the invention may also be administered in the form of a suppository in the rectal cavity. By combining the drug with a suitable non-irritating excipient, such as a synthetic glyceride of cocoa butter or polyethylene glycol, the composition is solid at room temperature but liquefied and/or dissolved in the rectal cavity. To release the drug.
由於個體受試者可能在症狀的嚴重程度呈現廣泛變化,而每種藥物都有其獨特的治療特徵,本發明之組合物實施應由醫師確定受試者對治療的反應,並相應地改變劑量。 Since individual subjects may exhibit broad variations in the severity of the symptoms, and each drug has its own unique therapeutic profile, the composition of the present invention should be performed by the physician to determine the subject's response to the treatment and to vary the dosage accordingly. .
圖一為本發明中草藥組合物治療中風後腦部傷害程度評估-梗塞面積暨水腫程度分析。事先餵食C57BL/6小鼠溶媒(CMC助懸劑)(對照組,CT)或不同倍數人體劑量(二倍:每天每公斤1.2公克,2x;或五倍:每天每公斤3公克,5x)之本發明中草藥組合物一個月後,將小鼠進行中腦動脈梗塞手術,24小時後利用影像系統評估其(A及B)腦部梗塞面積與(C)水腫程度之變化。每組動物數為4-6隻。***代表P值小於0.001。 Figure 1 is an analysis of the degree of brain injury after treatment of stroke in Chinese herbal medicine composition - infarct size and edema degree analysis. Pre-fed C57BL/6 mouse vehicle (CMC suspending agent) (control group, CT) or different multiple human dose (two times: 1.2 grams per kilogram per day, 2x; or five times: 3 grams per kilogram per day, 5x) One month after the herbal composition of the present invention, the mice were subjected to middle cerebral artery infarction, and 24 hours later, the changes in the area of brain infarction (A and B) and (C) the degree of edema were evaluated using an imaging system. The number of animals in each group is 4-6. *** represents a P value of less than 0.001.
圖二為本發明中草藥組合物治療中風後腦部傷害程度評估--動物行為模式分析。事先餵食C57BL/6小鼠溶媒(CMC助懸劑)(對照組,CT)或不同倍數人體劑量(二倍:每天每公斤1.2公克,2x;或五倍:每天每公斤3公克,5x)之本發明中草藥組合物一個月後,將小鼠進行中腦動脈梗塞手術,分別於1小時及24小時後評估其神經缺陷行為之程度。每組動物數為5-10隻。 Figure 2 is an evaluation of the degree of brain injury after treatment of stroke in Chinese herbal medicine composition of the present invention--analysis of animal behavior patterns. Pre-fed C57BL/6 mouse vehicle (CMC suspending agent) (control group, CT) or different multiple human dose (two times: 1.2 grams per kilogram per day, 2x; or five times: 3 grams per kilogram per day, 5x) One month after the herbal composition of the present invention, the mice were subjected to middle cerebral artery infarction, and the degree of neurological deficit behavior was evaluated after 1 hour and 24 hours, respectively. The number of animals in each group is 5-10.
本發明之目的係提供一種中草藥組合物,其用於預防或改善血栓或血栓相關疾病之用途,以及其製備方法。其中,該中草藥組合物包含川芎,白芷,雞血藤,木賊,川紅花,綠茶或其組合。 It is an object of the present invention to provide a Chinese herbal medicine composition for use in preventing or ameliorating a thrombosis or a thrombosis-related disease, and a preparation method thereof. Wherein, the Chinese herbal medicine composition comprises Chuanxiong, Baiji, Spatholobus, Equisetum, Chuanhonghua, green tea or a combination thereof.
以下之實施例非為限定用途,僅用以呈現此發明之多種面向。 The following examples are not intended to be limiting, but merely to present various aspects of the invention.
實施例一、中草藥組合物之製備方法。 Example 1. Preparation method of Chinese herbal medicine composition.
將川芎10克與白芷10克一起蒸煮半小時成第一鍋煮液;將雞血藤30克與木賊10克煎煮二小時成第二鍋煮液;川紅花5克與綠茶10克加熱半小時成第三鍋煮液。將第一鍋,第二鍋,以及第三鍋煮液混合後,進入濃縮,噴霧乾燥成粉末。 10 grams of Chuanxiong and 10 grams of white peony are cooked for half an hour into the first pot of cooking liquid; 30 grams of spatholobus and 10 grams of squid are boiled for 2 hours into a second pot of cooking liquid; 5 grams of safflower and 10 grams of green tea are heated for half an hour. The third pot of cooking liquid. The first pot, the second pot, and the third pot of cooking liquor are mixed, concentrated, and spray dried to a powder.
實施例二、治療中風後腦部傷害程度評估。 Example 2: Evaluation of the degree of brain injury after stroke treatment.
小鼠發中腦動脈血管阻塞/再灌流中風模式。 The middle cerebral artery occlusion/reperfusion stroke pattern in mice.
將六周大之C57BL/6品系小鼠(體重20~25公克)以3%異氟醚(isofluran,溶於95% O2和5% CO2)氣體混合器麻醉並以2%異氟醚(isoflurane)維持麻醉。頸部皮膚由中線切開,找出右側總頸動脈(right common carotid artery)、外頸動脈(external carotid artery)、與內頸動脈 (internal carotid artery),將線栓經由外頸動脈插入至內頸動脈,即造成右側中大腦動脈(right middle cerebral artery)之阻塞(ischemia),然後縫合傷口,使老鼠自然甦醒。利用行為測試確認小鼠處於成功的單一側的腦部缺血。二十四小時後,將腦組織取下做進一步的分析。手術過程中老鼠體溫均維持在溫度37℃。 Six-week-old C57BL/6 strain mice (body weight 20-25 g) were anesthetized with a 3% isoflurane (isofluran, dissolved in 95% O 2 and 5% CO 2 ) gas mixer and 2% isoflurane (isoflurane) ) Maintain anesthesia. The skin of the neck is cut from the midline to find the right common carotid artery, the external carotid artery, and the internal carotid artery. The wire plug is inserted through the external carotid artery. The carotid artery, which causes the ischemia of the right middle cerebral artery, then sutures the wound, allowing the mouse to naturally wake up. Behavioral testing was used to confirm that the mouse was on a successful single side of the brain ischemia. Twenty-four hours later, brain tissue was removed for further analysis. The body temperature of the mice was maintained at 37 °C during the operation.
腦梗塞區域(infarct size)之測定。 Determination of the infarct size.
經二十四小時的血液再灌流後,使用chloral hydrate每公斤200毫克麻醉老鼠,於低溫及含氧下將大腦組織取出,將之冠狀切片成2毫米厚度。以2% 2,3,5-三苯基氯化四氮唑(2,3,5-triphenyltetrazolium chloride,TTC)在溫度37℃下染色,細胞若存活則會將此染劑代謝成紅色並累積在細胞內,反之死亡的細胞則呈現白色,30分鐘後,隨即放入10%甲醛溶液(formaldehyde solution)固定。隔天,腦切片用數位相機照相(Nikon Coolpix 5000),最後以影像分析軟體(Image-Pro Plus)計算腦梗塞體積之百分比(% infarction volume),本實驗未受損的對側半腦為控制組,並換算出腦細胞的存活率,以評估藥物治療缺血性腦梗塞引發腦部傷害之療效。A:同側腦半球(右腦)未受損面積;B:對側腦半球(左腦)面積;校準後的腦梗塞率:(B-A)/B×100%。 After 24 hours of blood reperfusion, 200 mg of anesthetized mice per kiloliter of chloral hydrate were used to remove the brain tissue under low temperature and oxygen, and the crown was sliced to a thickness of 2 mm. Dyeing with 2% 2,3,5-triphenyltetrazolium chloride (TTC) at 37 ° C, the cells will metabolize to red and accumulate if the cells survive. In the cells, the cells that died in the opposite direction showed white color, and after 30 minutes, they were fixed in a 10% formaldehyde solution. The next day, brain slices were photographed with a digital camera (Nikon Coolpix 5000), and finally the percentage of cerebral infarction volume (% infarction volume) was calculated by Image-Pro Plus. The contralateral hemisphere of the experiment was controlled. Group, and converted the survival rate of brain cells to evaluate the efficacy of drugs in the treatment of brain damage caused by ischemic cerebral infarction. A: undamaged area of the ipsilateral cerebral hemisphere (right brain); B: area of the contralateral hemisphere (left brain); cerebral infarction rate after calibration: (B-A) / B × 100%.
神經缺陷分級(neurological deficits)。 Neurological deficits.
為評估老鼠中風後運動神經受損的程度。分別在第一小時與第二十四小時的血液再灌流時,請旁觀者先判定小鼠的行為模式,再將小鼠處以其他的實驗。此分級制度詳述如下,若老鼠呈現較嚴重的等級,以較嚴重的等級分類。神經學檢查過程歷時3~5分鐘。實驗步驟為:首先溫和地握住 老鼠尾巴使其懸掛距離地面一公尺,觀察前掌彎曲的現象。若老鼠的兩前掌能均衡延展向地面,並且沒有觀察到其他的神經缺損現象,為正常的老鼠,表示0級。有腦梗塞的老鼠會將前肢一致性地往腦損傷半球的對側(contralateral)收縮;而前肢收縮隨著損傷的程度不同而改變,輕度的收縮如腕彎曲、肩內收和肘僵直。較嚴重收縮則造成腕、肘完全彎曲與肩的內收和向內旋轉。老鼠有上述程度的前肢收縮且沒有其他異常時,歸類為第1級。老鼠被放置於軟質塑膠塗覆的大紙(counter protection paper,Kimberly Clarke)上時,會以爪子緊緊抓住紙張。以手抓住老鼠的尾巴,從肩後施以力量直到老鼠的前肢滑動數英吋。依此步驟從不同的方向操作數次,正常或輕微中風的老鼠對於不同方向的拉力有相同的抵抗力,嚴重中風的老鼠對朝麻痺體側的橫向施力會減低抵抗力,歸類為第2級。接著讓老鼠自由活動,觀察其繞圈的行為,若老鼠往麻痺的體側移動,歸類為第3級。若老鼠喪失走路或困難於行走,歸類為第4級。 To assess the extent of motor nerve damage after stroke in mice. When the blood is reperfused for the first hour and the twenty-fourth hour, the bystander first determines the behavior pattern of the mouse, and then the mouse is subjected to other experiments. This grading system is detailed below. If mice present a more severe grade, they are classified by a more severe grade. The neurological examination process lasted 3 to 5 minutes. The experimental procedure is as follows: firstly, hold the mouse tail gently and hang it one meter away from the ground to observe the bending of the forefoot. If the two forefoot of the mouse can be extended to the ground in a balanced manner, and no other nerve defects are observed, it is a normal mouse, indicating level 0. Mice with cerebral infarction contract the forelimbs concentrically to the contralateral side of the brain injury hemisphere; while forelimb contractions vary with the degree of injury, such as wrist flexion, shoulder adduction, and elbow stiffness. More severe contraction causes the wrist and elbow to bend completely and the shoulder to adduct and rotate inward. When the mouse has the above-mentioned degree of forelimb contraction and no other abnormalities, the mouse is classified as the first level. When the mouse is placed on a counter-protection paper (Kimberly Clarke), the paper is held tightly by the claws. Grab the tail of the mouse with your hand and apply force from behind the shoulder until the forelimb of the mouse slides a few inches. According to this procedure, the operation is performed several times in different directions. Rats with normal or slight stroke have the same resistance to the pulling force in different directions. The lateral force exerted by the severely stroked mouse on the side of the paralysis body will reduce the resistance, and is classified as the first level 2. Then let the mouse move freely and observe its behavior around the circle. If the mouse moves to the side of the paralysis, it is classified as level 3. If the mouse loses walking or is difficult to walk, it is classified as level 4.
本實驗之給藥設計為預先口服給予小鼠不同劑量之本發明中草藥組合物一個月後再進行手術;實驗分組分為:(1)未進行手術控制組(sham group)、(2)進行手術但未給藥組、(3)給予低劑量本發明中草藥組合物組、(4)給予高劑量本發明中草藥組合物組觀察其腦部傷害面積大小及腦梗塞的各項指標。 The administration of the experiment was designed to be administered orally to mice in different doses of the Chinese herbal medicine composition of the present invention for one month before the operation; the experimental grouping was divided into: (1) no operation control group (sham group), (2) surgery However, the unadministered group, (3) the low dose of the Chinese herbal medicine composition group of the present invention, and (4) the high dose of the Chinese herbal medicine composition group of the present invention were observed for the brain injury area and the indicators of cerebral infarction.
實驗數據皆以平均值±標準誤差(mean±S.E.M.)表示,控制組與實驗組之間的差異,實驗的統計則以單因子變異數分析(one-way ANOVA)作統計分析。若有意義差別,再以紐柯檢定法(Newman-Keuls)比較各組間的差異性,若P值小於0.05則表示有意義的差別。 The experimental data were expressed as mean ± standard error (mean ± S. E. M.), the difference between the control group and the experimental group, and the experimental statistics were analyzed by one-way ANOVA. If there is a meaningful difference, then the Newman-Keuls method is used to compare the differences between the groups. If the P value is less than 0.05, it means a meaningful difference.
實驗結果發現,以每天每公斤1.2公克或每天每公斤3公克劑量餵食小鼠本發明中草藥組合物一個月後,可有效預防缺血性腦中風手術所帶來的腦部傷害(圖一),及改善中風所帶來之神經缺陷行為(圖二)。 The experimental results showed that feeding the Chinese herbal composition of the present invention to 1.2 gram per kilogram per day or 3 gram per kilogram per day can effectively prevent brain damage caused by ischemic stroke surgery (Fig. 1). And improve the neurological deficits caused by stroke (Figure 2).
為使此發明所屬技術領域中具有通常知識者得以了解製作以及使用這項技藝的方法,此發明已描述並已充分詳細舉例說明,然而,各式各樣的變體,修改或改進應被視為無異於此項發明之精神與範圍。 This invention has been described and illustrated in sufficient detail to enable a person of ordinary skill in the art to which this invention pertains. The various modifications, modifications, and improvements should be considered. It is no different from the spirit and scope of this invention.
本發明所屬技術領域中具有通常知識者易於理解並實現本發明之目的,並獲得先前所提到之結果及優點。本發明所使用之動物,材料以及生產它們的過程和方法乃代表最佳實施例,乃示例性質,而不作為限制本發明的範圍用途。本領域的技術人員與製作或使用此項技藝時所將產生之修改或其他用途皆涵蓋於本發明的精神內,並且由權利範圍所限定。 It is obvious to those skilled in the art that the present invention can readily understand and achieve the objects of the present invention and obtain the results and advantages mentioned. The animals, materials, and processes and methods for producing the same are representative of the preferred embodiments and are exemplary in nature and are not intended to limit the scope of the invention. Modifications or other uses that may occur to those skilled in the art and in the art of making or using the art are encompassed within the spirit of the invention and are defined by the scope of the invention.
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