苦參止痢膠囊對(duì)犬細(xì)小病毒病的治療作用
發(fā)布時(shí)間：2019-06-09 21:06
【摘要】：犬細(xì)小病毒病因其高傳染性和高死亡率,成為危害犬最大的疫病之一。由于用于輔助治療的各種化學(xué)藥物會(huì)帶來(lái)耐藥性等一系列問(wèn)題,使得研發(fā)新型高效、無(wú)毒無(wú)副作用的抗犬細(xì)小病毒病中藥制劑成為新課題�？鄥⒅沽∧z囊是根據(jù)中獸醫(yī)理論和衛(wèi)氣營(yíng)血辨證施治的原則,選用苦參、白芍、木香等組成的中藥復(fù)方膠囊制劑,具有清熱燥濕,涼血止痢的功效。本文通過(guò)細(xì)胞試驗(yàn)、動(dòng)物試驗(yàn)和臨床用藥來(lái)探討苦參止痢膠囊對(duì)犬細(xì)小病毒病的治療作用。試驗(yàn)一高效液相色譜法測(cè)定苦參止痢膠囊有效成分的含量采用反向高效液相色譜法測(cè)定苦參止痢膠囊中苦參堿、氧化苦參堿、芍藥苷、去氫木香內(nèi)酯和木香烴內(nèi)酯的含量。采用Phenemonex C18(250mm×4.6mm,5μm)色譜柱,苦參堿和氧化苦參堿用10mmol/L的醋酸胺水溶液(用氨水調(diào)節(jié)pH8)-甲醇梯度洗脫的流動(dòng)相,紫外檢測(cè)波長(zhǎng)為210nm,流速:1mL/min;芍藥苷用乙腈-0.1%磷酸溶液(14:86)的流動(dòng)相,紫外檢測(cè)波長(zhǎng)為230nm,流速:1mL/min;去氫木香內(nèi)酯和木香烴內(nèi)酯用甲醇-水(65:35)的流動(dòng)相,紫外檢測(cè)波長(zhǎng)為225nm,流速:0.65mL/min。結(jié)果苦參止痢膠囊中苦參堿為2.2mg/g、氧化苦參堿為1.05mg/g、芍藥苷為9.1ng/g和去氫木香內(nèi)酯為13.5mg/g,木香烴內(nèi)酯由于含量過(guò)低未測(cè)出。試驗(yàn)二苦參止痢膠囊在細(xì)胞上對(duì)犬細(xì)小病毒的作用研究為了觀察苦參止痢膠囊體外對(duì)犬細(xì)小病毒的作用,以FK8]細(xì)胞為載體,通過(guò)間接免疫熒光試驗(yàn)(IFA),用熒光素標(biāo)記進(jìn)入細(xì)胞的犬細(xì)小病毒,在熒光倒置顯微鏡下觀察,用ImageJ軟件分析熒光。結(jié)果顯示先加中藥再感染病毒組和中藥與病毒作用后感染組都能顯著減少FK81細(xì)胞中犬細(xì)小病毒的表達(dá)量,先加中藥再感染病毒組平均減少量為36%,中藥與病毒作用后感染組平均減少量為46%;先感染病毒再加中藥組FK81細(xì)胞中病毒表達(dá)量與陽(yáng)性組無(wú)顯著差異。表明苦參止痢膠囊溶液在細(xì)胞上能有效阻止犬細(xì)小病毒的吸附和侵入,對(duì)于在細(xì)胞內(nèi)復(fù)制的犬細(xì)小病毒無(wú)明顯抑制作用。試驗(yàn)三中西藥治療犬細(xì)小病毒病的臨床試驗(yàn)為了觀察苦參止痢膠囊對(duì)犬細(xì)小病毒病的治療作用,用水平感染的7只病犬作為動(dòng)物模型,分為中藥組、西藥組和陽(yáng)性組,分別為3只、3只和1只,1只健康犬為空白,用苦參止痢膠囊和西藥治療,通過(guò)臨床癥狀、病理解剖和組織病理學(xué)切片光鏡及電鏡觀察。結(jié)果顯示:苦參止痢膠囊和西藥都能控制犬細(xì)小病毒病,其中苦參止痢膠囊組沒(méi)有出現(xiàn)血痢的癥狀,組織病理學(xué)切片觀察腸道組織也沒(méi)有明顯的出血,并且維持了腸絨毛結(jié)構(gòu)的完整性,表明苦參止痢膠囊能抑制犬細(xì)小病毒引起的腸黏膜損傷和出血性病變,從而對(duì)犬細(xì)小病毒病達(dá)到治愈作用。試驗(yàn)四苦參止痢膠囊治療犬細(xì)小病毒病的擴(kuò)大臨床試驗(yàn)為了進(jìn)一步探討苦參止痢膠囊防治犬細(xì)小病毒病的效果。方法:利用動(dòng)物醫(yī)院門(mén)診收治的犬細(xì)小病毒病病例,采用純西藥治療與中西藥聯(lián)合治療的對(duì)比試驗(yàn),統(tǒng)計(jì)兩種藥的治愈率、治愈療程等情況。結(jié)果.:用純西藥治療的32只犬細(xì)小病毒病病例的治愈率為75%;應(yīng)用苦參止痢膠囊的中西藥聯(lián)合治療的32只犬細(xì)小病毒病病例的治愈率為87.5%;且中西藥聯(lián)合治療的平均治愈療程為5.32d,低于純西藥治療的平均治愈療6.29d。中西藥聯(lián)合治療組治愈犬平均止住血痢天數(shù)為3.86d,顯著低于西藥組的5.38d(P0.05)。結(jié)論:苦參止痢膠囊對(duì)犬細(xì)小病毒病有良好輔助治療作用。
[Abstract]:The cause of canine parvovirus is its high infectivity and high mortality, which is one of the most dangerous diseases of the dog. The novel high-efficiency, non-toxic and side-effect anti-canine parvovirus disease traditional Chinese medicine preparation has become a new problem due to a series of problems such as drug resistance and the like caused by various chemical drugs used for adjuvant treatment. Radix Sophorae Flavescentis anti-dysentery capsule is prepared from Radix Sophorae Flavescentis, Radix Paeoniae Alba, and Radix Aucklandiae, and has the effects of clearing away heat, eliminating dampness, and cold blood and stopping dysentery, and is prepared from Radix Sophorae Flavescentis, Radix Paeoniae Alba, and Radix Aucklandiae. In this paper, the therapeutic effect of Radix Sophorae Flavescentis on the canine parvovirus disease was discussed by cell test, animal test and clinical medication. A high performance liquid chromatography (HPLC) was used to determine the content of matrine, oxymatrine, Paeonia lactiflora, dehydrocostunolide and costunolide in the capsule of Radix Sophorae Flavescentis with high performance liquid chromatography. The mobile phase of a 10 mmol/ L aqueous solution of acetic acid (pH 8)-methanol gradient was used with a Phenemonex C18 (250 mm, 4.6 mm,5. mu.m) column, and a mobile phase with a wavelength of 210 nm and a flow rate of 1 mL/ min with an ultraviolet detection wavelength of 210 nm and a flow rate of 1 mL/ min; and a mobile phase of a paeonol-0.1% phosphoric acid solution (14:86), The ultraviolet detection wavelength is 230 nm, the flow rate is 1 mL/ min, the mobile phase of the dehydro-costunolide and the costunolide with methanol-water (65:35), the ultraviolet detection wavelength is 225nm, the flow rate is 0.65 mL/ min. Results The matrine was 2.2 mg/ g, the oxymatrine was 1.05 mg/ g, the peony root was 9.1 ng/ g and the dehydro-costunolide was 13.5 mg/ g, and the content of the costunolide was not measured. in ord to observe that effect of the radix sophorae flavescentis anti-dysentery capsule on the canine parvovirus in the cell, in order to observe the effect of the radix sophorae flavescentis anti-dysentery capsule on the canine parvovirus in vitro, the FK8] cell is taken as a carrier, and the canine parvovirus which enters the cell is labeled with a fluorescein by an indirect immunofluorescence test (IFA), The fluorescence was analyzed with ImageJ software under a fluorescence inversion microscope. The results showed that the expression of the canine parvovirus in the FK81 cells was significantly reduced by the combination of the traditional Chinese medicine reinfected group and the infection group of the traditional Chinese medicine and the virus, the average reduction of the first Chinese medicine reinfected virus group was 36%, and the average reduction of the infection group after the effect of the traditional Chinese medicine and the virus was 46%; No significant difference was found between the expression of the virus and the positive group of the virus plus the group of FK81 in the traditional Chinese medicine group. It is shown that the solution of the anti-dysentery capsule of the radix sophorae flavescentis can effectively prevent the adsorption and the invasion of the canine parvovirus, and has no obvious inhibiting effect on the canine parvovirus which is reproduced in the cell. In order to observe the therapeutic effect of Radix Sophorae Flavescentis anti-dysentery capsule on canine parvovirus disease, the three-and-western medicine in the treatment of canine parvovirus disease was used as animal model, and it was divided into three groups: the traditional Chinese medicine group, the western medicine group and the positive group. 3,3 and 1,1 healthy dog were blank, and were treated with flavescent sophora root, antipullorum and western medicine, and were observed by light and electron microscope with clinical symptoms, pathological anatomy and tissue pathology. The results showed that both the anti-dysentery capsule and the western medicine of the radix sophorae flavescentis can control the canine parvovirus disease, wherein the radix sophorae flavescentis antidiarrheal capsule group does not have the symptoms of the blood pullorum, the pathological sections of the tissue do not have obvious bleeding in the intestinal tissue, and the integrity of the intestinal villi structure is maintained, It is shown that the anti-diarrhea capsule of the radix sophorae flavescentis can inhibit the intestinal mucosa injury and the hemorrhagic lesion caused by the canine parvovirus, thereby achieving the cure effect on the canine parvovirus disease. In order to further study the effect of Radix Sophorae Flavescentis for preventing and treating canine parvovirus disease, the clinical trial was conducted to study the effect of Radix Sophorae Flavescentis for preventing and treating canine parvovirus disease. Methods: In the case of canine parvovirus disease treated by the outpatient department of the animal hospital, the comparison between the treatment of pure western medicine and the combination of the Chinese and Western medicine was adopted, and the cure rate and the course of treatment of the two kinds of drugs were counted. As a result. the cure rate of 32 canine parvovirus disease cases treated with pure western medicine is 75%, the cure rate of 32 canine parvovirus disease cases treated with the combination of the Chinese and western medicines of the radix sophorae flavescentis anti-diarrhea capsule is 87.5%, and the average curing course of the combination therapy of the Chinese and western medicines is 5.32d, The mean cure was 6.29 d, which was lower than that of pure western medicine. The average number of patients cured by the combination of Chinese and Western medicine was 3.86d, which was significantly lower than that of the western medicine group (P0.05). Conclusion: The anti-dysentery capsule of Radix Sophorae Flavescentis has good adjuvant treatment effect on canine parvovirus disease.
【學(xué)位授予單位】：揚(yáng)州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S858.292
本文編號(hào)：2495900
[Abstract]:The cause of canine parvovirus is its high infectivity and high mortality, which is one of the most dangerous diseases of the dog. The novel high-efficiency, non-toxic and side-effect anti-canine parvovirus disease traditional Chinese medicine preparation has become a new problem due to a series of problems such as drug resistance and the like caused by various chemical drugs used for adjuvant treatment. Radix Sophorae Flavescentis anti-dysentery capsule is prepared from Radix Sophorae Flavescentis, Radix Paeoniae Alba, and Radix Aucklandiae, and has the effects of clearing away heat, eliminating dampness, and cold blood and stopping dysentery, and is prepared from Radix Sophorae Flavescentis, Radix Paeoniae Alba, and Radix Aucklandiae. In this paper, the therapeutic effect of Radix Sophorae Flavescentis on the canine parvovirus disease was discussed by cell test, animal test and clinical medication. A high performance liquid chromatography (HPLC) was used to determine the content of matrine, oxymatrine, Paeonia lactiflora, dehydrocostunolide and costunolide in the capsule of Radix Sophorae Flavescentis with high performance liquid chromatography. The mobile phase of a 10 mmol/ L aqueous solution of acetic acid (pH 8)-methanol gradient was used with a Phenemonex C18 (250 mm, 4.6 mm,5. mu.m) column, and a mobile phase with a wavelength of 210 nm and a flow rate of 1 mL/ min with an ultraviolet detection wavelength of 210 nm and a flow rate of 1 mL/ min; and a mobile phase of a paeonol-0.1% phosphoric acid solution (14:86), The ultraviolet detection wavelength is 230 nm, the flow rate is 1 mL/ min, the mobile phase of the dehydro-costunolide and the costunolide with methanol-water (65:35), the ultraviolet detection wavelength is 225nm, the flow rate is 0.65 mL/ min. Results The matrine was 2.2 mg/ g, the oxymatrine was 1.05 mg/ g, the peony root was 9.1 ng/ g and the dehydro-costunolide was 13.5 mg/ g, and the content of the costunolide was not measured. in ord to observe that effect of the radix sophorae flavescentis anti-dysentery capsule on the canine parvovirus in the cell, in order to observe the effect of the radix sophorae flavescentis anti-dysentery capsule on the canine parvovirus in vitro, the FK8] cell is taken as a carrier, and the canine parvovirus which enters the cell is labeled with a fluorescein by an indirect immunofluorescence test (IFA), The fluorescence was analyzed with ImageJ software under a fluorescence inversion microscope. The results showed that the expression of the canine parvovirus in the FK81 cells was significantly reduced by the combination of the traditional Chinese medicine reinfected group and the infection group of the traditional Chinese medicine and the virus, the average reduction of the first Chinese medicine reinfected virus group was 36%, and the average reduction of the infection group after the effect of the traditional Chinese medicine and the virus was 46%; No significant difference was found between the expression of the virus and the positive group of the virus plus the group of FK81 in the traditional Chinese medicine group. It is shown that the solution of the anti-dysentery capsule of the radix sophorae flavescentis can effectively prevent the adsorption and the invasion of the canine parvovirus, and has no obvious inhibiting effect on the canine parvovirus which is reproduced in the cell. In order to observe the therapeutic effect of Radix Sophorae Flavescentis anti-dysentery capsule on canine parvovirus disease, the three-and-western medicine in the treatment of canine parvovirus disease was used as animal model, and it was divided into three groups: the traditional Chinese medicine group, the western medicine group and the positive group. 3,3 and 1,1 healthy dog were blank, and were treated with flavescent sophora root, antipullorum and western medicine, and were observed by light and electron microscope with clinical symptoms, pathological anatomy and tissue pathology. The results showed that both the anti-dysentery capsule and the western medicine of the radix sophorae flavescentis can control the canine parvovirus disease, wherein the radix sophorae flavescentis antidiarrheal capsule group does not have the symptoms of the blood pullorum, the pathological sections of the tissue do not have obvious bleeding in the intestinal tissue, and the integrity of the intestinal villi structure is maintained, It is shown that the anti-diarrhea capsule of the radix sophorae flavescentis can inhibit the intestinal mucosa injury and the hemorrhagic lesion caused by the canine parvovirus, thereby achieving the cure effect on the canine parvovirus disease. In order to further study the effect of Radix Sophorae Flavescentis for preventing and treating canine parvovirus disease, the clinical trial was conducted to study the effect of Radix Sophorae Flavescentis for preventing and treating canine parvovirus disease. Methods: In the case of canine parvovirus disease treated by the outpatient department of the animal hospital, the comparison between the treatment of pure western medicine and the combination of the Chinese and Western medicine was adopted, and the cure rate and the course of treatment of the two kinds of drugs were counted. As a result. the cure rate of 32 canine parvovirus disease cases treated with pure western medicine is 75%, the cure rate of 32 canine parvovirus disease cases treated with the combination of the Chinese and western medicines of the radix sophorae flavescentis anti-diarrhea capsule is 87.5%, and the average curing course of the combination therapy of the Chinese and western medicines is 5.32d, The mean cure was 6.29 d, which was lower than that of pure western medicine. The average number of patients cured by the combination of Chinese and Western medicine was 3.86d, which was significantly lower than that of the western medicine group (P0.05). Conclusion: The anti-dysentery capsule of Radix Sophorae Flavescentis has good adjuvant treatment effect on canine parvovirus disease.
【學(xué)位授予單位】：揚(yáng)州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：S858.292
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