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1.中国中医科学院 中药研究所 生物安全实验室,北京 100700
2.武汉健民大鹏药业有限公司,武汉 430000
赵荣华,助理研究员,从事中药药理抗病毒抗菌药效机制研究,E-mail:rhzhao@icmm.ac.cn
时宇静,副研究员,从事中药药理抗病毒机制研究,E-mail:yjshi@icmm.ac.cn
纸质出版日期:2021-01-20,
网络出版日期:2020-11-23,
收稿日期:2020-10-23,
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赵荣华,孙静,郭姗姗等.体外培育牛黄对人冠状病毒肺炎疫毒袭肺证小鼠病证结合模型的效用特点[J].中国实验方剂学杂志,2021,27(02):66-73.
ZHAO Rong-hua,SUN Jing,GUO Shan-shan,et al.Evaluation Effectiveness of in Vitro Cultivation of Bezoar on Mouse Model Combining Disease with Syndrome of Coronavirus Pneumonia with Yidu Xifei Syndrome[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(02):66-73.
赵荣华,孙静,郭姗姗等.体外培育牛黄对人冠状病毒肺炎疫毒袭肺证小鼠病证结合模型的效用特点[J].中国实验方剂学杂志,2021,27(02):66-73. DOI: 10.13422/j.cnki.syfjx.20210143.
ZHAO Rong-hua,SUN Jing,GUO Shan-shan,et al.Evaluation Effectiveness of in Vitro Cultivation of Bezoar on Mouse Model Combining Disease with Syndrome of Coronavirus Pneumonia with Yidu Xifei Syndrome[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(02):66-73. DOI: 10.13422/j.cnki.syfjx.20210143.
目的
2
研究药物—体外培育牛黄对人冠状病毒肺炎疫毒袭肺证动物模型的药理作用特点。
方法
2
按体质量等级将48只Balb/c小鼠随机分为6组,分别为空白组,人冠状病毒229E感染组,寒湿组,人冠状病毒肺炎疫毒袭肺复合模型组,体外培育牛黄高、低剂量组,每组8只。采用寒湿环境诱导+人冠状病毒229E感染形成复合动物病证结合模型,在感染当天给予体外培育牛黄(0.128,0.064 g‧kg
-1
),灌胃给药,连续3 d,观察小鼠一般状态,小鼠肺指数和肺指数抑制率;采用实时荧光定量聚合酶链式反应(Real-time PCR)检测小鼠肺组织病毒载量;采用酶联免疫吸附测定(ELISA)检测小鼠血清胃动素(MTL),胃泌素(GAS)含量,以及小鼠肺组织细胞因子白细胞介素(IL)-10,IL-6,肿瘤坏死因子(TNF)-
α
及
γ-
干扰素(IFN-
γ
)含量;采用流式细胞术检测小鼠血中CD4
+
T淋巴细胞,CD8
+
T淋巴细胞及B淋巴细胞。
结果
2
体外培育牛黄高、低剂量组可明显改善模型小鼠的一般状态,与空白组比较,模型组小鼠肺指数显著升高(
P
<
0.01),小鼠肺组织核酸表达显著升高(
P
<
0.01),小鼠血清中MTL含量显著增高,GAS含量明显降低(
P
<
0.05,
P
<
0.01),肺部组织细胞免疫因子TNF-
α
,IL-10和IL-6均显著增高(
P
<
0.01),小鼠外周血淋巴细胞CD4
+
T细胞,CD8
+
T细胞及B细胞均显著降低(
P
<
0.01);与模型组比较,体外培育牛黄高、低剂量组小鼠肺指数均显著降低(
P
<
0.01),小鼠肺组织核酸表达显著降低(
P
<
0.01),MTL含量显著降低(
P
<
0.01),小鼠肺组织中IL-6,IL-10,TNF-
α
,IFN-
γ
含量均显著降低(
P
<
0.01),体外培育牛黄高剂量组可显著升高CD4
+
T细胞(
P
<
0.05),体外培育牛黄可一定程度减轻模型小鼠肺组织炎性渗出,肺间质水肿及瘀血等病理表现。
结论
2
体外培育牛黄对人冠状病毒感染致疫毒袭肺证小鼠复合模型的药理作用有明显的改善效用,可显著改善小鼠行为及排泄物状态、降低模型小鼠肺指数、提高肺指数抑制率、降低小鼠肺组织核酸表达、减轻肺组织病理损伤、调节免疫机能和抑制炎性因子释放等作用环节发挥作用,为临床用药提供依据。
Objective
2
To determine the therapeutic effect of
in vitro
cultivation of bezoar on a mouse model adding disease with syndrome of coronavirus pneumonia with Yidu Xifei syndrome.
Method
2
BALB/c mice were randomly divided into six groups according to their weight grade: normal group, HCoV-229E infection group, cold and damp group, a mouse model combining disease with syndrome of coronavirus pneumonia with Yidu Xifei syndrome, and high and low dose group of
in vitro
cultivation of bezoar. The combination model of human coronavirus pneumonia with Yidu Xifei syndrome mice was established by the method of cold dampness condition stimulation+coronavirus HCoV-229E infection.
In vitro
cultivation of bezoar (0.128,0.064 g·kg
-1
) was administrated by gavage for 3 days from the day of infection. The observation indexes included: general state observation of mice, inhibition rate of lung index and lung index of mice. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the viral load in the lung tissues of mice. Serum levels of motilin(MTL), gastrin (GAS), and cytokines interleukin(IL)-10,IL-6, tumor necrosis factor-
α
(TNF-
α
)and interferon-
γ
(IFN-
γ
) in lung tissue of mice were determined by enzyme-linked immunosorbent assay(ELISA). The percentages of CD4
+
T lymphocytes,CD8
+
T lymphocytes and B lymphocytes in the blood of mice were determined by flow cytometry.
Result
2
The high and low dose group of
in vitro
cultivation of bezoar can significantly improve the general condition of model mice. Compared with blank group, model group mice lung index increased significantly (
P
<
0.01), nucleic acids significantly increased expression of lung tissue in mice (
P
<
0.01), significantly higher serum MTL content in mice, GAS content significantly decreased (
P
<
0.05,
P
<
0.01), lung tissue cells in the immune factor TNF-
α
, IL-10 and IL-6 were significantly increased (
P
<
0.01), peripheral blood lymphocyte CD4
+
T cells in mice, The percentages of CD8
+
T cells and B cells were significantly decreased (
P
<
0.01). Compared with model group,
in vitro
cultivation bezoar mice lung index of high and low dose group were significantly lower (
P
<
0.01), the lung tissue of mice express nucleic acid decreased significantly (
P
<
0.01), MTL content decreased significantly (
P
<
0.01), the lung tissue of mice in the IL-6, IL-10, the TNF-
α
, IFN-
γ
levels were significantly lower (
P
<
0.01),
in vitro
cultivation bezoar high dose group can significantly increase the CD4
+
T cell percentage (
P
<
0.05),
in vitro
cultivation bezoar can to a certain extent reduce model mice lung inflammatory exudation, pulmonary interstitial edema, as well as blood stasis symptoms.
Conclusion
2
In vitro
cultivation of bezoar has a significant therapeutic effect on a mice model adding disease with syndrome of coronavirus pneumonia with Yidu Xifei syndrome. It can be treated by reducing the lung index of the model mice, improving the pathological damage of the lung tissue, adjusting the immune effective and inhibiting the clearing of inflammatory factors, and to provide a laboratory basis for clinical medication.
新型冠状病毒肺炎体外培育牛黄中医病机疫毒袭肺证免疫机能炎性因子
corona virus disease-2019in vitro cultivation of bezoarpathogenesis of traditional Chinese medicineYidu Xifei syndromeimmune functioninflammatory factors
国家卫生健康委员会,国家中医药管理局.《新型冠状病毒肺炎诊疗方案(试行第七版)》解读[EB/OL]. http://www.nhc.gov.cn/yzygj/s7652m/202003/a31191442e29474b98bfed5579d5af95.shtmlhttp://www.nhc.gov.cn/yzygj/s7652m/202003/a31191442e29474b98bfed5579d5af95.shtml,2020-03-03/2020-03-04.
北京市中医管理局.北京市新型冠状病毒感染的肺炎防治方案(第二版)[EB/OL].http://zyj.beijing.gov.cn/sy/tzgg/202001/t20200130_1621630.htmlhttp://zyj.beijing.gov.cn/sy/tzgg/202001/t20200130_1621630.html,2020-01-29/2020-02-28.
石岩,孙冬梅,熊婧,等.人工牛黄多组分测定及其质量差异标志物的研究[J].中国中药杂志,2018,43(4):659-664..
刘灵,白丽,陈慧婷.牛黄对慢阻肺大鼠抗炎、抗凋亡的实验研究[J].光明中医,2018,33(6):795-798.
刘小雪,王莹,陈鹏,等.体外培育牛黄联合甲泼尼龙琥珀酸钠注射剂治疗难治性支原体肺炎的临床观察[J].中国中西医结合杂志,2019,39(8):960-964.
LYU X,MA Y,WU F,et al.LncRNA NKILA inhibits retinoblastoma by downregulating lncRNA XIST[J].Curr Eye Res,2019,44(9),975-979.
AURELIO B,BRUCE D Z,DAVID E W,et al. Identification of a receptor-binding domain of the spike glycoprotein of human coronavirus HCoV-229E[J].J Virol,2003,77(4):2530-2538.
WILLINGER T,FLAVELL R A.Canonical autophagy dependent on the class Ⅲ phosphoinositide-3 kinase Vps34 is required for naive T-cell homeostasis[J].Proc Natl Acad Sci USA,2012,109(22):8670-8675.
吴伟,温敏勇,詹少锋,等.基于中医瘟疫火热病机探讨新型冠状病毒肺炎辨证论治[J].中国中西医结合杂志,2020,40(3):272-274.
陈光敏.新型冠状病毒性肺炎的病因病位病机、治法方药[J].中医临床研究,2020,12(8):24-27.
范逸品,张华敏,王燕平,等.新型冠状病毒肺炎中医疾病属性归类简析[J].中医杂志,2020,61(11):921-927.
王琦,谷晓红,刘清泉.新型冠状病毒肺炎中医诊疗手册[M].北京:中国中医药出版社,2020:9-12.
胡家光,蒋忠胜,李旭,等.新型冠状病毒肺炎患者16例外周血T淋巴细胞亚群的变化及意义[J].广东医学,2020,41(8):781-783.
宋霞,陈涛,孙晶晶,等.PCT与IL-6联合检测对新冠病毒感染肺炎的临床价值探讨[J].国际检验医学杂志,2020,dio:50.1176.R.20200309.1926.002.
张益明,高世定,王占科,等.3例新型冠状病毒肺炎患者外周血CD4+ T淋巴细胞和CD4+/CD8+比值变化[J].国际检验医学杂志,2020,41(17):2173-2176.
何秀玲,李培锋,关红,等.牛磺酸鹅去氧胆酸对小鼠免疫功能的影响[J].中药材,2005,28(12):1089-1092.
贺雯茜,张程亮,向东,等.基于血清药理学技术研究体外培育牛黄抑制肝细胞脂质沉积的作用机制[J].中国中药杂志,2019,44(17):3780-3785.
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