锦灯笼有效成分抗炎作用实验研究
|
摘要
咽炎是影响人类健康和生活质量的疾病之一。急、慢性咽炎所导致的病理改变,不仅造成咽局部的机能障碍,也可波及邻近器官组织,甚至影响全身其它系统,损害人体的健康。现代社会生活、工作压力的增加,空气环境的污染,尤其在我国东北地区,气候干燥,咽炎的发病率有升高的趋势。目前临床上治疗各种急、慢性咽炎的药物大多为抗生素类药物,近年来由于微生物对抗生素耐药越来越严重,这使抗生素类药物的临床应用收到限制。中药治疗咽炎具有毒副作用低、作用持久的特点,近年来迅速成为医学和药学研究者的关注热点。
本课题研究选用盛产于长白山区的锦灯笼,经水提醇沉有机溶剂萃取得到粗提物(Extracts of Physalis alkekengi L var. franchetii (Mast.) Makino, EPAFM)。
采用体内外实验方法,分别从毒理学、药效学和免疫学等方面研究了EPAFM对小鼠的体内毒性、体内外抗菌活性、抗炎活性、祛痰活性和免疫调节活性。实验结果表明:EPAFM对小鼠的MTD>20g/kg。EPAFM体外抗金黄色葡萄球菌和乙型溶血性链球菌的活性较高,其中对乙型溶血性链球菌的抗菌活性要高于金黄色葡萄球菌。EPAFM体内能有效抑制大鼠咽炎模型的诱发菌的生长。EPAFM能减轻二甲苯所致的炎性水肿,降低咽炎动物外周血中炎细胞数量,减少炎症部位炎细胞的浸润,抑制纤维组织增生。小鼠酚红实验结果表明,EPAFM有一定的祛痰效果。EPAFM对免疫器官重量和T淋巴细胞转化能力影响不大,但对血清抗体IgG有很强的促进作用。提示:EPAFM可通过抗菌、降低毛细血管通透性、提高血清免疫球蛋白含量发挥其治疗咽炎的作用。
The acute and chronic pharyngitis is a throat familiar and frequently-occurring disease. Pathological Changes induced by pharyngitis can not merely cause dysfunction of thoat, but affect peripheral organs,function, even the whole body, It has a bad influence on human health. At present, drugs for treatment pharyngitis mainly in clinical application are of antibiotics, but they have lower effect and drug resistances. Consequently, it is urgent to find new drugs that have high performance and low toxicities. Traditional Chinese Medicine poses low toxicities and persistent action, becoming the hot spot of researching new drugs against pharyngitis.
Physalis alkekengi L var.franchetii (Mast.) Makino has an effect of heatclearing and detoxifying, often used to treat throat gall and hoarseness. In this thesis, we got extraction from Physalis alkekengi L var.franchetii (Mast.) Makino (EPAFM) in Changbai Mountains by means of water extracting, alcohol precipitation and organic solvent extracting. Toxicities, anti- inflammation activity and reducing sputum function, immunoregulatory activity were measured in vitro and in vivo.
1. Toxicity of EPAFM in vivo : the toxicity of EPAFM in vivo was measured by the method of maximum tolerated dose testing. The result showed that maximal tolerance dose of EPAFM is higher than 20g/kg.
2. Antimicrobial activity of EPAFM in vitro and in vivo: Antimicrobial activity of EPAFM in vitro and in vivo are separately measured by paper piece method and bacterial culture of pharyngo swab method. The results showed that EPAFM is sensitive to S . aureus and str.Hemolyticus in vitro,and high dose group of EPAFM has a better inhibition than medium、low dose goups and watermelon frost control group.
3. Anti-inflammatory activity of EPAFM: The anti-inflammatory effect of EPAFM was evaluated using the model of dimethylbenzene-induced mouse ear swelling and rat model of acute bacterial Pharyngitis. The results showed that EPAFM can significantly decrease the ear swelling degree and alleviate inflammation. In the rat model of acute bacterial Pharyngitis, The decrease of white blood cell counts in peripheral blood and inflammatory cells in the mucosa were observed. Hint: EPAFM exerted significant antiinflammatory activity
4. Expectorant function of EPAFM: Expectorant experiment was conducted with methods of washing mice's respiratory tract with phenol red. The result showed that EPAFM has a nice expectorant function at the dose of 2.1g/kg.
5. Immunoregulatory activities of EPAFM: Immunoregulatory activities of EPAFM were measured by the methods of organ weighter, T Lymphocyte transformation test and Level of Serum IgG. The results showed that the stimulation on immune organs proliferation and promotion of T Lymphocyte transformation function is not obvious, while its promotion of Serum IgG production.
Conlusion In summary, our results indicate that the MTD of EPAFM is higher than 20g/kg. EPAFM has a nice effect of antimicrobial activity, which can effevtively inhibit the growth and propogation of many bacteria. It also has a anti-inflammation effect , restraining permeability of capillary pressure and decreasing inflammatory cells an the lesion. EPAFM plays a role immunomodulation, it can increase the content of IgG in serum and activate anti-infection immune response.
本课题研究选用盛产于长白山区的锦灯笼,经水提醇沉有机溶剂萃取得到粗提物(Extracts of Physalis alkekengi L var. franchetii (Mast.) Makino, EPAFM)。
采用体内外实验方法,分别从毒理学、药效学和免疫学等方面研究了EPAFM对小鼠的体内毒性、体内外抗菌活性、抗炎活性、祛痰活性和免疫调节活性。实验结果表明:EPAFM对小鼠的MTD>20g/kg。EPAFM体外抗金黄色葡萄球菌和乙型溶血性链球菌的活性较高,其中对乙型溶血性链球菌的抗菌活性要高于金黄色葡萄球菌。EPAFM体内能有效抑制大鼠咽炎模型的诱发菌的生长。EPAFM能减轻二甲苯所致的炎性水肿,降低咽炎动物外周血中炎细胞数量,减少炎症部位炎细胞的浸润,抑制纤维组织增生。小鼠酚红实验结果表明,EPAFM有一定的祛痰效果。EPAFM对免疫器官重量和T淋巴细胞转化能力影响不大,但对血清抗体IgG有很强的促进作用。提示:EPAFM可通过抗菌、降低毛细血管通透性、提高血清免疫球蛋白含量发挥其治疗咽炎的作用。
The acute and chronic pharyngitis is a throat familiar and frequently-occurring disease. Pathological Changes induced by pharyngitis can not merely cause dysfunction of thoat, but affect peripheral organs,function, even the whole body, It has a bad influence on human health. At present, drugs for treatment pharyngitis mainly in clinical application are of antibiotics, but they have lower effect and drug resistances. Consequently, it is urgent to find new drugs that have high performance and low toxicities. Traditional Chinese Medicine poses low toxicities and persistent action, becoming the hot spot of researching new drugs against pharyngitis.
Physalis alkekengi L var.franchetii (Mast.) Makino has an effect of heatclearing and detoxifying, often used to treat throat gall and hoarseness. In this thesis, we got extraction from Physalis alkekengi L var.franchetii (Mast.) Makino (EPAFM) in Changbai Mountains by means of water extracting, alcohol precipitation and organic solvent extracting. Toxicities, anti- inflammation activity and reducing sputum function, immunoregulatory activity were measured in vitro and in vivo.
1. Toxicity of EPAFM in vivo : the toxicity of EPAFM in vivo was measured by the method of maximum tolerated dose testing. The result showed that maximal tolerance dose of EPAFM is higher than 20g/kg.
2. Antimicrobial activity of EPAFM in vitro and in vivo: Antimicrobial activity of EPAFM in vitro and in vivo are separately measured by paper piece method and bacterial culture of pharyngo swab method. The results showed that EPAFM is sensitive to S . aureus and str.Hemolyticus in vitro,and high dose group of EPAFM has a better inhibition than medium、low dose goups and watermelon frost control group.
3. Anti-inflammatory activity of EPAFM: The anti-inflammatory effect of EPAFM was evaluated using the model of dimethylbenzene-induced mouse ear swelling and rat model of acute bacterial Pharyngitis. The results showed that EPAFM can significantly decrease the ear swelling degree and alleviate inflammation. In the rat model of acute bacterial Pharyngitis, The decrease of white blood cell counts in peripheral blood and inflammatory cells in the mucosa were observed. Hint: EPAFM exerted significant antiinflammatory activity
4. Expectorant function of EPAFM: Expectorant experiment was conducted with methods of washing mice's respiratory tract with phenol red. The result showed that EPAFM has a nice expectorant function at the dose of 2.1g/kg.
5. Immunoregulatory activities of EPAFM: Immunoregulatory activities of EPAFM were measured by the methods of organ weighter, T Lymphocyte transformation test and Level of Serum IgG. The results showed that the stimulation on immune organs proliferation and promotion of T Lymphocyte transformation function is not obvious, while its promotion of Serum IgG production.
Conlusion In summary, our results indicate that the MTD of EPAFM is higher than 20g/kg. EPAFM has a nice effect of antimicrobial activity, which can effevtively inhibit the growth and propogation of many bacteria. It also has a anti-inflammation effect , restraining permeability of capillary pressure and decreasing inflammatory cells an the lesion. EPAFM plays a role immunomodulation, it can increase the content of IgG in serum and activate anti-infection immune response.
引文
1 叶京英,韩德民.慢性咽炎研究进展[J].中国医学文摘·耳鼻咽喉科学,2004,19(5):268
2 康厚墉,谭斌.慢性咽炎中西医结合治疗近况[J].中西医结合杂志,2003,11(3):154
3 江苏新医学院.中药大辞典[M],上海:上海科技出版社,1977:2532-2533
4 王明东, 杨松松. 锦灯笼化学成分及药理作用综述[J].辽宁中医学院学报, 2005, 7(4): 341-342
5 李娟,李静,李德坤. 锦灯笼化学成分的研究(Ⅱ) [J]. 中草药, 2002,33(9):788-789
6 李娟,李静,李德坤. 锦灯笼化学成分的研究(Ⅰ) [J]. 中草药, 2002,33(8):692-693
7 Mstusura T, Kawai M, Nakashima R, et al. Bitter principles of physalis alkekengi L.Var.franchetii;Structure of Physalin A [J]. Tetrahedron lett, 1969, 14, 1083一1086.
8 M.Kawai, T.Matsuura, S.Kyuno, et al. A new physalin from physalis alkekengi structure of physain L [J]. Phyto-chemistry,1987, 26:3313-3317
9 M. Kawai, T. Ogura, M. Nakamishi, et al. Structure of Physalin isolated from physalis alkekengi Var. Francheti [J]. Bull Chem. Soc. Jpn, 1988,261(7): 2696-2698
10 M. Kawai, A. Matsumoto, B. Makino, et al. The structure of physalin P, a new physalin from physalis alkekengi[J]. Bull.Chem.Sco.Jpn,1993, 66:1299-1301
11 L. R. Row, N,S. Sarma, T. Matsuura, and R. Nakashima. Physalins E and H, new physalins from physalis angulata and physalis lancifolia[J]. Phytochemistry,1993,17:1641-1643
12 L. R. Row, N,S. Sarma, K. S. Reddy, N,S. Sarma, T. Matsuura,R. Nakashima. New physalins from physalis angulata and physalis lancifolia structure and reactions of physalins D,I,G and K. Phytochemistry[J]. 1980,19:1175-1175
13 Partha Neogi, Mahendra Sahai , Anil B. Ray. Withaperuvins F and G, two withanolides of physalis peruviona roots[J]. Phytochemistry, 1987, 26(1): 243-245
14 郑虎占. 中药现代研究与应用(第六卷)[M], 学苑出版社, 1999: 5885-5889
15 周小平. 酸浆果实与宿萼无机元素的比较分析[J]. 白求恩医科大学学报,2001, 27(4): 363-384
16 杨晓虹, 田丁丁, 周小平等. 酸浆宿萼无机元素和氨基酸含量的测定[J]. 白求恩医科大学学报,2001, 27 (1 ) :17-18
17 《全国中草药汇编》编写组.全国中草药汇编(上册)[M].1976:886
18 中华人民共和国卫生部药典委员会. 中华人民共和国药典(2005 年版一部. 2005: 250-251
19 北京市卫生局编,中草药制剂技术[M],化学工业出版社,1978:352
20 北京中医院儿科.新医药学杂志,1973,(1):17-19.
21 刘清贞,崔文成,王延泉等.山东中医杂志[J],1997,16(8):344-345
22 李萍,盛巡,王晓中等. 酸浆苦素B对活化多形核中性粒细胞化学发光及H20:产生的影响[J].中草药1996,27(15):284-286
23 李萍,盛巡,王晓中等. 酸浆苦素B对活化多性核中性细胞化学发光及H20:产生的影响[J].中国病理生理杂志,1996,12(6):626-627.
24 李萍,盛巡,王晓中等. 活化嗜中性粒细胞[Ca2+]的变化及锦灯笼酸浆苦素B的影响[J].中国病理生理杂志,1996,12(6):660-662
25 宋立人.现代中药学大词典(下册)[M].北京:人民卫生出版社,1999:2311-2314
26 袁昌衡,周启贵,杨飞中.80 种中药水煎液对淋球菌的抑制试验[J].国医院药学杂志,1997,17(11):508-509
27 何丽娜,何素冰,杨军. 木犀草素体外抗柯萨奇B3病毒作用[J]. 中国现代应用药学杂,2000,17(5):362 -365
28 刘勇明编著.维吾尔药志(下册)[M].新疆:新疆科技卫生出版社,1999:918-922
29 龚珊,单立冬. 挂金灯镇痛作用的实验观察[J].苏州大学学报(医学版), 2002,22(4):380-382
30 王和平,徐美术,孙亮等.锦灯笼降血糖作用的实验研究[J]. 中医药息,2004,21(1):53-54
31 李德坤. 锦灯笼化学成分的研究.吉林大学硕士学位论文.2001
32 Antoun MD, Abramson D, Tyson RL, et al. Potential antitumor agents. XVII. Physalin B and 25,26-epidihydrophysalin C from Witheringia coccoloboides[J]. J Nat Prod 1981,44:579-585
33 Han R.Recent progress in the study of anti-cancer drugs originating from plants and traditional medicines in China[J].Chin Med Sci J,1994,9:61-69
34 Hartwell JL.Plants used against cancer.A survey.Lawrence,CA:Quarterman Publications, 1982
35 Dornberger K.The potential antineoplastic acting constituents of Physalisalkekengi L. var franchetii Mast. Pharmazie,1986,41(4):265-268.
36 刘寿山. 中药文献研究摘要(1920-1961)[M]. 1963: 742
37 伊津野吉亮.日本药理学杂志,1957, 53:400-403
38 Vessal M, Mehrani HA, Omrani GH.Effects of an aqueous extract of Physalis alkekengi fruit on estrus cycle, reproduction and uterine creatine kinase BB-isozymein rats[J].Ethnopharmacol,1991,34(1):69-78
39 张昊.灯笼草对致敏小鼠肝损伤的保护作用.延边大学医院.2007
40 李仪奎主编.中药药理实验方法学[M].上海:上海科学技术出版社,1991:299-310
41 徐叔云、卞如濂、陈修主编.药理实验方法学[M].北京:人民卫生出版社,1992:719
42 龚守良主编.实用基础医学实验技术[M].长春:吉林科学技术出版社,1991:170-171
43 王迪浔. 病理生理学[M]. 北京:人民卫生出版社,1994:109
44 高学敏. 中药学[M]. 北京:人民卫生出版社,2000
45 杨春燕, 杨兴海, 刘英等. 开口箭祛痰、抗炎及抑菌实验研究[J]. 中国民族民间医药杂志, 2005, 73: 102-105
46 陈慰峰. 医学免疫学(第三版)[M], 2001:9-12
47 Janeway CA, et al. Immunobiology,4th ed, chapter 8. Current Biology Publications, 1999:263-305
48 杨贵贞主编. 医学免疫学(第五版)[M]. 北京:高等教育出版 社,2003:68
49 李良庆.玉屏风合施尔康对小儿呼吸道感染免疫球蛋白的影响[J]. 现 代中西医结合杂志,2000, 9(18):1797
50 Avanzini MA, Bjorkander J, Soderstrom R, et al. Qualitative and quantitative analyses of the antibody response elicited by Haemophilusinfluenza type b capsular polysaccharide-tetanus toxoid conjugates in adults with IgG subclass deficiencies and frequent infections [ J ]. Clin Exp Immunol, 1994, 96 (1) : 540 - 581
51 Kapellerova, Kasanicra A, Ponee J, et al. IgG subclasses in children with recurrent respiratory diseases[ J ]. Cesk Pediatr, 1993, 48 (12) :697-700
2 康厚墉,谭斌.慢性咽炎中西医结合治疗近况[J].中西医结合杂志,2003,11(3):154
3 江苏新医学院.中药大辞典[M],上海:上海科技出版社,1977:2532-2533
4 王明东, 杨松松. 锦灯笼化学成分及药理作用综述[J].辽宁中医学院学报, 2005, 7(4): 341-342
5 李娟,李静,李德坤. 锦灯笼化学成分的研究(Ⅱ) [J]. 中草药, 2002,33(9):788-789
6 李娟,李静,李德坤. 锦灯笼化学成分的研究(Ⅰ) [J]. 中草药, 2002,33(8):692-693
7 Mstusura T, Kawai M, Nakashima R, et al. Bitter principles of physalis alkekengi L.Var.franchetii;Structure of Physalin A [J]. Tetrahedron lett, 1969, 14, 1083一1086.
8 M.Kawai, T.Matsuura, S.Kyuno, et al. A new physalin from physalis alkekengi structure of physain L [J]. Phyto-chemistry,1987, 26:3313-3317
9 M. Kawai, T. Ogura, M. Nakamishi, et al. Structure of Physalin isolated from physalis alkekengi Var. Francheti [J]. Bull Chem. Soc. Jpn, 1988,261(7): 2696-2698
10 M. Kawai, A. Matsumoto, B. Makino, et al. The structure of physalin P, a new physalin from physalis alkekengi[J]. Bull.Chem.Sco.Jpn,1993, 66:1299-1301
11 L. R. Row, N,S. Sarma, T. Matsuura, and R. Nakashima. Physalins E and H, new physalins from physalis angulata and physalis lancifolia[J]. Phytochemistry,1993,17:1641-1643
12 L. R. Row, N,S. Sarma, K. S. Reddy, N,S. Sarma, T. Matsuura,R. Nakashima. New physalins from physalis angulata and physalis lancifolia structure and reactions of physalins D,I,G and K. Phytochemistry[J]. 1980,19:1175-1175
13 Partha Neogi, Mahendra Sahai , Anil B. Ray. Withaperuvins F and G, two withanolides of physalis peruviona roots[J]. Phytochemistry, 1987, 26(1): 243-245
14 郑虎占. 中药现代研究与应用(第六卷)[M], 学苑出版社, 1999: 5885-5889
15 周小平. 酸浆果实与宿萼无机元素的比较分析[J]. 白求恩医科大学学报,2001, 27(4): 363-384
16 杨晓虹, 田丁丁, 周小平等. 酸浆宿萼无机元素和氨基酸含量的测定[J]. 白求恩医科大学学报,2001, 27 (1 ) :17-18
17 《全国中草药汇编》编写组.全国中草药汇编(上册)[M].1976:886
18 中华人民共和国卫生部药典委员会. 中华人民共和国药典(2005 年版一部. 2005: 250-251
19 北京市卫生局编,中草药制剂技术[M],化学工业出版社,1978:352
20 北京中医院儿科.新医药学杂志,1973,(1):17-19.
21 刘清贞,崔文成,王延泉等.山东中医杂志[J],1997,16(8):344-345
22 李萍,盛巡,王晓中等. 酸浆苦素B对活化多形核中性粒细胞化学发光及H20:产生的影响[J].中草药1996,27(15):284-286
23 李萍,盛巡,王晓中等. 酸浆苦素B对活化多性核中性细胞化学发光及H20:产生的影响[J].中国病理生理杂志,1996,12(6):626-627.
24 李萍,盛巡,王晓中等. 活化嗜中性粒细胞[Ca2+]的变化及锦灯笼酸浆苦素B的影响[J].中国病理生理杂志,1996,12(6):660-662
25 宋立人.现代中药学大词典(下册)[M].北京:人民卫生出版社,1999:2311-2314
26 袁昌衡,周启贵,杨飞中.80 种中药水煎液对淋球菌的抑制试验[J].国医院药学杂志,1997,17(11):508-509
27 何丽娜,何素冰,杨军. 木犀草素体外抗柯萨奇B3病毒作用[J]. 中国现代应用药学杂,2000,17(5):362 -365
28 刘勇明编著.维吾尔药志(下册)[M].新疆:新疆科技卫生出版社,1999:918-922
29 龚珊,单立冬. 挂金灯镇痛作用的实验观察[J].苏州大学学报(医学版), 2002,22(4):380-382
30 王和平,徐美术,孙亮等.锦灯笼降血糖作用的实验研究[J]. 中医药息,2004,21(1):53-54
31 李德坤. 锦灯笼化学成分的研究.吉林大学硕士学位论文.2001
32 Antoun MD, Abramson D, Tyson RL, et al. Potential antitumor agents. XVII. Physalin B and 25,26-epidihydrophysalin C from Witheringia coccoloboides[J]. J Nat Prod 1981,44:579-585
33 Han R.Recent progress in the study of anti-cancer drugs originating from plants and traditional medicines in China[J].Chin Med Sci J,1994,9:61-69
34 Hartwell JL.Plants used against cancer.A survey.Lawrence,CA:Quarterman Publications, 1982
35 Dornberger K.The potential antineoplastic acting constituents of Physalisalkekengi L. var franchetii Mast. Pharmazie,1986,41(4):265-268.
36 刘寿山. 中药文献研究摘要(1920-1961)[M]. 1963: 742
37 伊津野吉亮.日本药理学杂志,1957, 53:400-403
38 Vessal M, Mehrani HA, Omrani GH.Effects of an aqueous extract of Physalis alkekengi fruit on estrus cycle, reproduction and uterine creatine kinase BB-isozymein rats[J].Ethnopharmacol,1991,34(1):69-78
39 张昊.灯笼草对致敏小鼠肝损伤的保护作用.延边大学医院.2007
40 李仪奎主编.中药药理实验方法学[M].上海:上海科学技术出版社,1991:299-310
41 徐叔云、卞如濂、陈修主编.药理实验方法学[M].北京:人民卫生出版社,1992:719
42 龚守良主编.实用基础医学实验技术[M].长春:吉林科学技术出版社,1991:170-171
43 王迪浔. 病理生理学[M]. 北京:人民卫生出版社,1994:109
44 高学敏. 中药学[M]. 北京:人民卫生出版社,2000
45 杨春燕, 杨兴海, 刘英等. 开口箭祛痰、抗炎及抑菌实验研究[J]. 中国民族民间医药杂志, 2005, 73: 102-105
46 陈慰峰. 医学免疫学(第三版)[M], 2001:9-12
47 Janeway CA, et al. Immunobiology,4th ed, chapter 8. Current Biology Publications, 1999:263-305
48 杨贵贞主编. 医学免疫学(第五版)[M]. 北京:高等教育出版 社,2003:68
49 李良庆.玉屏风合施尔康对小儿呼吸道感染免疫球蛋白的影响[J]. 现 代中西医结合杂志,2000, 9(18):1797
50 Avanzini MA, Bjorkander J, Soderstrom R, et al. Qualitative and quantitative analyses of the antibody response elicited by Haemophilusinfluenza type b capsular polysaccharide-tetanus toxoid conjugates in adults with IgG subclass deficiencies and frequent infections [ J ]. Clin Exp Immunol, 1994, 96 (1) : 540 - 581
51 Kapellerova, Kasanicra A, Ponee J, et al. IgG subclasses in children with recurrent respiratory diseases[ J ]. Cesk Pediatr, 1993, 48 (12) :697-700