板蓝根超微粉对大鼠免疫功能调节作用的研究
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摘要
为研究板蓝根经超微粉碎后对大鼠免疫功能调节的作用影响,利用环磷酰胺构建大鼠免疫低下模型,将80只SD大鼠随机分为4组,即模型组、普通粉组、超微粉组及空白对照组,每组20只。40 mg/kg环磷酰胺腹腔注射5 d制造免疫抑制大鼠模型,第6天开始给药,除空白组、模型组灌服生理盐水外,其他组给药5 g/kg。第7、14天采集血样及大鼠肝脏、脾脏、胸腺组织,测定免疫器官指数、白细胞数量、C3b受体花环形成率(C3b-R)、红细胞免疫复合物花环形成率(ICR)、吞噬百分率、吞噬指数等指标,实时荧光定量PCR检测肿瘤坏死因子α(TNF-α)、白细胞介素2(IL-2) mRNA转录水平,Western blotting法检测TNF-ɑ和IL-2蛋白表达水平。结果显示,模型组大鼠白细胞数量、体重、胸腺和脾脏免疫器官指数较空白对照组均显著降低(P<0.05)。第14天时,超微粉组巨噬细胞吞噬百分率达到54.75%,吞噬指数为0.58;C3b-R为9.44%,显著高于普通粉组(8.97%)(P<0.05),超微粉组ICR为6.09%,低于普通粉组(6.33%)(P>0.05)。与普通粉组相比,第14天板蓝根超微粉组肝脏中TNF-α、IL-2 mRNA转录水平和蛋白水平的表达提高。综上所述,板蓝根超微粉和普通粉均可显著增强大鼠非特异性免疫功能,但超微粉效果优于普通粉。
In order to study the effects of ultrafine powder of Radix Isatidis on immune regulation in rats,cyclophosphamide was used to construct the model of immunodeficiency.80 SD rats were randomly divided into 4 groups:Model group,ordinary powder group,ultrafine powder group and blank control group with 20 rats in each group.40 mg/kg cyclophosphamide was injected intraperitoneally for 5 d.From 6 th day,The blank control group and model group were given normal saline,and the other groups were given 5 g/kg ordinary powder or ultrafine powder.At 7 th and 14 th day,the blood samples and liver,spleen and thymus tissues were collected.The white blood cell count(WBC),organ indexes of spleen and thymus,C3 b-R,ICR,phagocytosis indexes were detected.Real-time PCR was used to detect the transcription levels of TNF-α and IL-2 mRNA.The expression of TNF-ɑ and IL-2 protein were detected by Western blotting.The results showed that the WBC,body weight,thymus and spleen indexes in model group were significantly lower than those in the blank control group(P<0.05).On the 14 th day,the percentage of macrophage phagocytosis of rat in ultrafine powder group reached 54.7%,the index of macrophage phagocytosis was 0.58.The C3 b-R of rat at 14 th day in ultrafine powder group was 9.44%,significantly higher than that of the ordinary power group(8.97%)(P<0.05).And the ICR of ultrafine powder group was 6.09%,lower than that of the ordinary power group(6.33%)(P>0.05).Compared with the ordinary powder group,the ultrafine powder of Radix Isatidis could increase the expression of TNF-ɑ and IL-2 in liver on the 14 th day.In conlusion,ordinary powder and ultrafine powder of Radix Isatidis could both enhance the nonspecific immune function of rats,and ultrafine powder was superior to the ordinary powder.
In order to study the effects of ultrafine powder of Radix Isatidis on immune regulation in rats,cyclophosphamide was used to construct the model of immunodeficiency.80 SD rats were randomly divided into 4 groups:Model group,ordinary powder group,ultrafine powder group and blank control group with 20 rats in each group.40 mg/kg cyclophosphamide was injected intraperitoneally for 5 d.From 6 th day,The blank control group and model group were given normal saline,and the other groups were given 5 g/kg ordinary powder or ultrafine powder.At 7 th and 14 th day,the blood samples and liver,spleen and thymus tissues were collected.The white blood cell count(WBC),organ indexes of spleen and thymus,C3 b-R,ICR,phagocytosis indexes were detected.Real-time PCR was used to detect the transcription levels of TNF-α and IL-2 mRNA.The expression of TNF-ɑ and IL-2 protein were detected by Western blotting.The results showed that the WBC,body weight,thymus and spleen indexes in model group were significantly lower than those in the blank control group(P<0.05).On the 14 th day,the percentage of macrophage phagocytosis of rat in ultrafine powder group reached 54.7%,the index of macrophage phagocytosis was 0.58.The C3 b-R of rat at 14 th day in ultrafine powder group was 9.44%,significantly higher than that of the ordinary power group(8.97%)(P<0.05).And the ICR of ultrafine powder group was 6.09%,lower than that of the ordinary power group(6.33%)(P>0.05).Compared with the ordinary powder group,the ultrafine powder of Radix Isatidis could increase the expression of TNF-ɑ and IL-2 in liver on the 14 th day.In conlusion,ordinary powder and ultrafine powder of Radix Isatidis could both enhance the nonspecific immune function of rats,and ultrafine powder was superior to the ordinary powder.
引文
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[10] LI Z,LI L,ZHOU H,et al.Radix Isatidis polysaccharides inhibit influenza a virus and influenza a virus-induced inflammation via suppression of host TLR3 signaling in vitro[J].Molecules,2017,22(1):116.
[11] 黄其春,何玉琴,李焰,等.银杏叶提取物的免疫调节作用研究进展[J].中国药理学通报,2010,26(2):278.HUANG Q C,HE Y Q,LI Y,et al.Advance in immuomodulatory effect of Ginkgo biloba extract[J].Chinese Pharmacological Bulletin,2010,26(2):278.(in Chinese)
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[13] 康慧琳,樊卫平,雷波,等.不同剂量环磷酰胺对小鼠免疫功能的影响[J].免疫学杂志,2018,34(4):308-312.KANG K L,FAN W P,LEI B.et al.The immunosuppression and immunoenhancement effects of cyclophosphamide on normal mice[J].Immunological Journal,2018,34(4):308-312.(in Chinese)
[14] 许桂红.板蓝根多糖化学与溶液黏度及免疫调节活性的研究[D].扬州:扬州大学,2006.XU G H.Study on chemical and solution viscosity and immunomodulatory activity of Radix Isatidis[D].Yangzhou:Yangzhou University,2006.(in Chinese)
[15] 张俊,胡安君,毕亚楠,等.板蓝根多糖对环磷酰胺造模大鼠的双向免疫调节作用[J].药物评价研究,2016,39(4):531-535.ZHANG J,HU A J,BI Y N,et al.Dual-directional immune regulation of Radix Isatidis polysaccharides on rats treated with cytoxan[J].Drug Evaluation Research,2016,39(4):531-535.(in Chinese)
[16] 李秀萍.硒化淫羊藿多糖和硒化板蓝根多糖增强免疫和抗氧化活性的研究[D].南京:南京农业大学,2015.LI X P.Antioxidant and immune-enhancing activities of selenium epimedium polysaccharide and selenizing Isatis root polysaccharide[D].Nanjing:Nanjing Agricutural University,2015.(in Chinese)
[17] 史自涛,姚焰础,江山,等.粪肠球菌替代抗生素对断奶仔猪生长性能、腹泻率、血液生化指标和免疫器官的影响[J].动物营养学报,2015,27(6):1832-1840.SHI Z T,YAO Y H,JIANG S,et al.Effects of Enterococcus faecalis substitute for antibiotic on growth performance,diarrhea rate,blood biochemical parameters and immune organs of weaner piglets [J].Chinese Journal of Animal Nutrition,2015,27(6):1832-1840.(in Chinese)
[18] 赵现敏,崔保安,张红英,等.4种中药多糖对猪脾淋巴细胞增殖的影响[J].河北农业大学学报,2007,30(3):82-85.ZHAO X M,CUI B A,ZHANG H Y,et al.Effects of four Chinese herbal medicinal polysaccharides on proliferation of swine splenic lymphocytes[J].Journal of Agricutural University of Hebei,2007,30(3):82-85.(in Chinese)
[19] 耿婵娟,刘志明,迟晓星,等.板蓝根粗多糖对免疫抑制小鼠的免疫调节作用[J].农产品加工,2012,4:36-39.GENG C J,LIU Z M,CHI X X,et al.Immunoregulatory effects of Isatis indigotica fortune raw polysaccharide on immunosuppressed mice[J].Academic Periodical of Farm Products Processing,2012,4:36-39.(in Chinese)
[20] 赵泽军,王志旺,郭玫,等.甘肃产不同生态型板蓝根对小鼠抗炎、免疫调节作用的比较[J].中国应用生理杂志,2018,34(1):57-60.ZHAO Z J,WANG Z W,GUO M,et al.Comparative study on anti-inflammatory and immunomodulation of different Radix Isatidis strains growing in Gansu province[J].Chinese Journal of Applied Physiology,2018,34(1):57-60.(in Chinese)
[2] 郭妍婷,黄雪,陈曼,超微粉碎技术的应用研究进展[J].广东化工,2016,43(16):276-277.GUO Y T,HUANG X,CHEN M,et al.Progress in research on the use of superfine grinding technology[J].Guangdong Chemical Industry,2016,43(16):276-277.(in Chinese)
[3] 赵国巍,梁新丽,廖正根,等.超微粉碎技术对中药粉体性质的影响[J].世界中医药,2015,3:315-317.ZHAO G W,LIANG X L,LIAO Z G.Effect of superfine grinding on properties of traditional Chinese medicine powder[J].World Chinese Medicine,2015,3:315-317.(in Chinese)
[4] 马毅敏,李娜,刘承伟,等.板蓝根不同提取部位抗炎镇痛活性比较研究[J].中草药,2014,45(17):2517-2521.MA Y M,LI N,LIU C W,et al.Comparative studies on anti-inflammatory and analgesic activities of different fractions from Radix Isatidis[J].Chinese Traditional and Herbal Drugs,2014,45(17):2517-2521.(in Chinese)
[5] 谭敏洁,江勋,彭婷婷,等.复方板蓝根颗粒质量标准提升研究[J].中国药业,2018,27(15):20-23.TAN M J,JIANG X,PENG T T,et al.Study on improvement of quality standard of compound Banlangen granules[J].China Pharmaceuticals,2018,27(15):20-23.(in Chinese)
[6] LIU Y H,LI J,XIE W,et al.Anti-endotoxic effects of o-aminobenzoic acid from Radix Isatidis[J].Central South Pharmacy,2005,26(5):593-597.
[7] LIU Y H,FENG J G,LI T,et al.Anti-endotoxic effects of syringic acid of Radix Isatidis[J].Journal of Huazhong University of Science and Technology:Medical Sciences,2003,23(2):206-208.
[8] 马丽娜,章从恩,鄢丹,等.超滤质谱技术筛选板蓝根中抗流感病毒的活性成分[J].中国中药杂志,2014,39(5):812-816.MA L N,ZHANG C E,YAN D,et al.Screening bioactive compounds inhibiting influenza virus from Isatidis Radix by ultrafiltration mass spectrometry [J].China Journal of Chinese Materia Medica,2014,39(5):812-816.(in Chinese)
[9] LIU Z,TANG Y,ZHOU R,et al.Bi-directional solid fermentation products of Trametes robiniophila Murr with Radix Isatidis inhibit proliferation and metastasis of breast cancer cells[J].Journal of the Chinese Medical Association,2018,81(6):520-530.
[10] LI Z,LI L,ZHOU H,et al.Radix Isatidis polysaccharides inhibit influenza a virus and influenza a virus-induced inflammation via suppression of host TLR3 signaling in vitro[J].Molecules,2017,22(1):116.
[11] 黄其春,何玉琴,李焰,等.银杏叶提取物的免疫调节作用研究进展[J].中国药理学通报,2010,26(2):278.HUANG Q C,HE Y Q,LI Y,et al.Advance in immuomodulatory effect of Ginkgo biloba extract[J].Chinese Pharmacological Bulletin,2010,26(2):278.(in Chinese)
[12] 魏恒,李娟娟.中药免疫增强剂在养鸡生产中的应用研究进展[J].山西农业科学,2018,46(11):1971-1974.WEI H,LI J J.Research progress of application of traditional Chinese medicine immunopotentiator in raising chicken[J].Journal of Shanxi Agricultural Sciences,2018,46(11):1971-1974.(in Chinese)
[13] 康慧琳,樊卫平,雷波,等.不同剂量环磷酰胺对小鼠免疫功能的影响[J].免疫学杂志,2018,34(4):308-312.KANG K L,FAN W P,LEI B.et al.The immunosuppression and immunoenhancement effects of cyclophosphamide on normal mice[J].Immunological Journal,2018,34(4):308-312.(in Chinese)
[14] 许桂红.板蓝根多糖化学与溶液黏度及免疫调节活性的研究[D].扬州:扬州大学,2006.XU G H.Study on chemical and solution viscosity and immunomodulatory activity of Radix Isatidis[D].Yangzhou:Yangzhou University,2006.(in Chinese)
[15] 张俊,胡安君,毕亚楠,等.板蓝根多糖对环磷酰胺造模大鼠的双向免疫调节作用[J].药物评价研究,2016,39(4):531-535.ZHANG J,HU A J,BI Y N,et al.Dual-directional immune regulation of Radix Isatidis polysaccharides on rats treated with cytoxan[J].Drug Evaluation Research,2016,39(4):531-535.(in Chinese)
[16] 李秀萍.硒化淫羊藿多糖和硒化板蓝根多糖增强免疫和抗氧化活性的研究[D].南京:南京农业大学,2015.LI X P.Antioxidant and immune-enhancing activities of selenium epimedium polysaccharide and selenizing Isatis root polysaccharide[D].Nanjing:Nanjing Agricutural University,2015.(in Chinese)
[17] 史自涛,姚焰础,江山,等.粪肠球菌替代抗生素对断奶仔猪生长性能、腹泻率、血液生化指标和免疫器官的影响[J].动物营养学报,2015,27(6):1832-1840.SHI Z T,YAO Y H,JIANG S,et al.Effects of Enterococcus faecalis substitute for antibiotic on growth performance,diarrhea rate,blood biochemical parameters and immune organs of weaner piglets [J].Chinese Journal of Animal Nutrition,2015,27(6):1832-1840.(in Chinese)
[18] 赵现敏,崔保安,张红英,等.4种中药多糖对猪脾淋巴细胞增殖的影响[J].河北农业大学学报,2007,30(3):82-85.ZHAO X M,CUI B A,ZHANG H Y,et al.Effects of four Chinese herbal medicinal polysaccharides on proliferation of swine splenic lymphocytes[J].Journal of Agricutural University of Hebei,2007,30(3):82-85.(in Chinese)
[19] 耿婵娟,刘志明,迟晓星,等.板蓝根粗多糖对免疫抑制小鼠的免疫调节作用[J].农产品加工,2012,4:36-39.GENG C J,LIU Z M,CHI X X,et al.Immunoregulatory effects of Isatis indigotica fortune raw polysaccharide on immunosuppressed mice[J].Academic Periodical of Farm Products Processing,2012,4:36-39.(in Chinese)
[20] 赵泽军,王志旺,郭玫,等.甘肃产不同生态型板蓝根对小鼠抗炎、免疫调节作用的比较[J].中国应用生理杂志,2018,34(1):57-60.ZHAO Z J,WANG Z W,GUO M,et al.Comparative study on anti-inflammatory and immunomodulation of different Radix Isatidis strains growing in Gansu province[J].Chinese Journal of Applied Physiology,2018,34(1):57-60.(in Chinese)