
浏览全部资源
扫码关注微信
1.甘肃中医药大学 甘肃省中药新产品创制工程实验室,甘肃省中医方药挖掘与创新转化重点实验室,兰州 730000
2.兰州大学 第一医院,兰州 730013
戴丽蓉,硕士,讲师,从事中西医结合基础研究,E-mail:549635490@qq.com
陈启明,硕士,主治医师,从事中西医结合重症医学研究,E-mail:81355875@163.com
收稿日期:2021-11-02,
网络出版日期:2022-03-28,
纸质出版日期:2022-06-05
移动端阅览
戴丽蓉,陈启明,刘喜平等.半夏泻心汤及其拆方对菌群紊乱幼鼠结肠黏膜免疫的影响[J].中国实验方剂学杂志,2022,28(11):42-50.
DAI Li-rong,CHEN Qi-ming,LIU Xi-ping,et al.Banxia Xiexintang and Its Disassembled Prescriptions Regulate Colonic Mucosal Immunity in Young Rats with Flora Disorder[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(11):42-50.
戴丽蓉,陈启明,刘喜平等.半夏泻心汤及其拆方对菌群紊乱幼鼠结肠黏膜免疫的影响[J].中国实验方剂学杂志,2022,28(11):42-50. DOI: 10.13422/j.cnki.syfjx.20221038.
DAI Li-rong,CHEN Qi-ming,LIU Xi-ping,et al.Banxia Xiexintang and Its Disassembled Prescriptions Regulate Colonic Mucosal Immunity in Young Rats with Flora Disorder[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(11):42-50. DOI: 10.13422/j.cnki.syfjx.20221038.
目的
2
探讨半夏泻心汤及其拆方对混合抗生素诱导的菌群紊乱幼鼠的治疗作用及可能的作用机制。
方法
2
将70只雄性BALB/c幼鼠随机分为7组,分别为空白组、模型组、双歧杆菌四联活菌片组(西药组,0.68 g·kg
-1
)及半夏泻心汤全方组、辛开组、苦降组、甘补组(9.1、3.19、1.82、4.1 g·kg
-1
),每组10只。除空白组外,其余各组予混合抗生素灌胃诱导肠道菌群紊乱,14 d后西药组予双歧杆菌四联活菌片灌胃,全方组、辛开组、苦降组及甘补组分别予半夏泻心汤及不同配伍药组灌胃,空白组、模型组给予等体积生理盐水,1次/d,共14 d。连续给药14 d后无菌采集粪便样本用于16SrDNA测序分析肠道菌群,并经腹腔注射脂多糖(LPS,10 mg·kg
-1
)诱导炎性反应,苏木素-伊红(HE)染色观察结肠黏膜组织形态,甲苯胺蓝染色和免疫组织化学观察结肠黏膜巨噬细胞浸润,实时荧光定量聚合酶链式反应(Real-time PCR)检测白细胞介素-1
β
(IL-1
β
)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-
α
(TNF-
α
)及白细胞介素-10(IL-10) mRNA的表达情况。
结果
2
与空白组比较,模型组幼鼠肠道微生物结构显著改变(
P
<
0.01),结肠黏膜损伤明显,巨噬细胞浸润减少,炎性因子IL-1
β
、IL-6、IL-8、TNF-
α
及IL-10 mRNA表达水平均显著降低(
P
<
0.01)。与模型组比较,西药组、半夏泻心汤及其拆方组均能增加菌群相对丰度和物种多样性,提高拟杆菌门、厚壁菌门等有益菌的比例(
P
<
0.01);同时西药组、半夏泻心汤及其拆方组均能改善结肠黏膜损伤(
P
<
0.05,
P
<
0.01),增加巨噬细胞浸润(
P
<
0.05,
P
<
0.01),升高IL-6、IL-8、TNF-
α
mRNA表达水平(
P
<
0.01);西药组、半夏泻心汤及苦降、甘补组升高IL-1
β
mRNA表达(
P
<
0.01);西药组、半夏泻心汤及辛开、甘补组IL-10 mRNA表达升高(
P
<
0.05,
P
<
0.01)。
结论
2
半夏泻心汤及拆方组能调整抗生素暴露幼鼠肠道菌群,保护肠道菌群紊乱后结肠黏膜免疫屏障,以半夏泻心汤全方为最佳配伍。
Objective
2
To explore the therapeutic effect and possible mechanism of Banxia Xiexintang and its disassembled prescriptions in regulating the flora disorder induced by mixed antibiotics in young rats.
Method
2
Seventy male BALB/C young rats were randomly assigned into 7 groups: blank group, model group, Bifidobacterium tetralogy viable tablets (0.68 g·kg
-1
) group, Banxia Xiexintang (9.1 g·kg
-1
) group, Xinkai (3.19 g·kg
-1
) group, Kujiang (1.82 g·kg
-1
) group, and Ganbu (4.1 g·kg
-1
) group, with 10 rats in each group. Except the blank group, the other groups were given mixed antibiotics by gavage to induce intestinal flora disorder. After 14 days, the rats in different drug groups were administrated with corresponding drugs by gavage, and those in the blank group and model group with the same amount of normal saline once a day for 14 days. After that, fecal samples were collected aseptically for 16S rDNA sequencing of intestinal flora, and lipopolysaccharide (LPS, 10 mg·kg
-1
) was injected intraperitoneally to induce inflammatory reaction. The tissue morphology of colonic mucosa was observed via hematoxylin-eosin (HE) staining, and the macrophage infiltration of colonic mucosa was observed via toluidine blue staining and immunohistochemistry. The expression of interleukin-1
β
(IL-1
β
), interleukin-6 (IL-6), interleukin-8 (IL-8), tumor necrosis factor-
α
(TNF-
α
), and interleukin-10 (IL-10) mRNA were determined by real-time fluorescence quantitative polymerase chain reaction (Real-time PCR).
Result
2
Compared with the blank group, the modeling changed the intestinal flora structure of the young rats (
P
<
0.01), damaged the colonic mucosa, reduced the macrophage infiltration, and down-regulated the mRNA levels of IL-1
β
, IL-6, IL-8, TNF-
α
, and IL-10 (
P
<
0.01). Compared with the model group, bifidobacterium quadruple viable tablets, Banxia Xiexintang and its disassembled prescriptions increased the diversity of intestinal flora and the relative abundance of beneficial bacteria such as Bacteroidetes and Firmicutes (
P
<
0.01). At the same time, they ameliorated colonic mucosal injury (
P
<
0.05,
P
<
0.01), increased macrophage infiltration (
P
<
0.05,
P
<
0.01), and up-regulated the mRNA levels of IL-6, IL-8, and TNF-
α
(
P
<
0.01). The mRNA level of IL-1
β
was up-regulated in Bifidobacterium tetralogy viable tablets, Banxia Xiexintang, Kujiang, and Ganbu groups (
P
<
0.01), and that of IL-10 was up-regulated in Bifidobacterium tetralogy viable tablets, Banxia Xiexintang, Xinkai, and Ganbu groups (
P
<
0.01).
Conclusion
2
Banxia Xiexintang and the disassembled prescriptions can adjust the intestinal flora of young rats exposed to antibiotics and protect the immune barrier of colonic mucosa after intestinal flora disorder. In particularly, the whole prescription of Banxia Xiexintang demonstrates the best performance.
詹妮 , 崔文静 , 金玉芬 , 等 . 肠道微生态失衡在代谢综合征发病机制中的研究 [J]. 中国实验诊断学 , 2021 , 25 ( 8 ): 1256 - 1259 .
郭宏伟 , 张妮妮 , 张伟 , 等 . 抗生素诱导的菌群紊乱对幼鼠结肠黏膜屏障及免疫反应的影响 [J]. 中华实用儿科临床杂志 , 2019 , 34 ( 7 ): 505 - 509 .
李玉丽 , 袁振仪 , 谭周进 . 泄泻证候相关肠道菌群的研究进展 [J]. 中国实验方剂学杂志 , 2021 , 27 ( 16 ): 209 - 217 .
张庚鑫 , 杜海洋 , 王平 , 等 . 基于16SrRNA测序研究葛根芩连汤对菌群失调性腹泻大鼠肠道菌群结构的影响 [J]. 中国实验方剂学杂志 , 2021 , 27 ( 11 ): 19 - 26 .
陈茹琴 . 半夏泻心汤合四逆散治疗菌群失调性肠炎12例 [J]. 中医杂志 , 2004 , 45 ( 1 ): 38 - 38 .
朱春亮 . 半夏泻心汤加黄芪治疗寒热错杂型腹泻型肠易激综合征的临床研究 [D]. 福州 : 福建中医药大学 , 2019 .
陈健 , 张梁坤 , 谷文超 , 等 . 半夏泻心汤对右旋葡聚糖硫酸钠诱导的溃疡性结肠炎小鼠肠道菌群的影响 [J]. 中国中药杂志 , 2021 , 46 ( 11 ): 2871 - 2880 .
崔国宁 , 刘喜平 , 王磊 , 等 . 半夏泻心汤治疗胃肠道疾病研究进展 [J]. 辽宁中医药大学学报 , 2019 , 21 ( 6 ): 87 - 90 .
张映红 , 戴丽蓉 , 陈启明 , 等 . 半夏泻心汤及不同配伍对抗生素诱导的菌群紊乱幼鼠结肠黏膜屏障的影响 [J]. 中国中医药信息杂志 , 2021 , doi: 10.19879/j.cnki.1005-5304.202110186 http://dx.doi.org/10.19879/j.cnki.1005-5304.202110186 .
邓中甲 . 方剂学 [M]. 北京 : 中国中医药出版社 , 2003 : 88 .
EDGAR R C . UPARSE:Highly accurate OTU sequences from microbial amplicon reads [J]. Nat Methods , 2013 , 10 ( 10 ): 996 - 998 .
WANG Q , GARRITY G M , TIEDJE J M , et al . Naive bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy [J]. Appl Environ Microbiol , 2007 , 73 ( 16 ): 5261 - 5267 .
张艳 . 半夏泻心汤及拆方对溃疡性结肠炎大鼠Th1/Th2细胞因子的影响 [D]. 青岛 : 山东中医药大学 , 2015 .
陈红苓 , 徐俊杰 . 肠道菌群变化对肠道免疫的影响及与小儿迁延性、慢性腹泻的关系 [J]. 中国妇幼保健 , 2019 , 34 ( 7 ): 1690 - 1692 .
王娟 , 范雅廷 , 董小英 . NICU新生儿院内感染危险因素和病原菌分布情况 [J]. 中国妇幼保健 , 2021 , 36 ( 5 ): 1136 - 1139 .
陈茹琴 . 半夏泻心汤合四逆散治疗菌群失调性肠炎12例 [J]. 中医杂志 , 2004 , 45 ( 1 ): 38 - 38 .
黄彦平 , 詹达法 , 黄海 , 等 . 半夏泻心汤人含药血清对HP感染GES-1细胞TGF- β /Smad信号通路的影响 [J]. 中国实验方剂学杂志 , 2018 , 24 ( 14 ): 91 - 96 .
LU W W , FU T X , WANG Q , et al . The effect of total glucoside of paeony on gut microbiota in NOD mice with Sjogren's syndrome based on high-throughput sequencing of 16SrRNA gene [J]. Chin Med , 2020 , 15 ( 7 ): 61 .
SHIN N R , WHON T W , BAE J W . Proteobacteria:Microbial signature of dysbiosis in gut microbiota [J]. Trends Biotechnol , 2015 , 33 ( 9 ): 496 - 503 .
YE M , JOOSSE M E , LIU L , et al . Deletion of IL-6 exacerbates colitis and induces systemic inflammation in IL-10-deficient mice [J]. J Crohns Colitis , 2020 , 14 ( 6 ): 831 - 840 .
杜小梅 , 潘薇 , 梁颖兰 , 等 . 参芪地黄汤加减治疗气阴两虚型糖尿病肾病疗效观察及对肠道菌群和炎症因子的影响 [J]. 中药新药与临床药理 , 2021 , 32 ( 4 ): 566 - 572 .
0
浏览量
18
下载量
5
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010802024621