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1.南京中医药大学 药学院,南京 210023
2.中国中医科学院 中药资源中心,北京 100700
3.中国中医科学院 中药研究所,北京 100700
4.安徽中医药大学 药学院,合肥 230012
席蕴文,在读硕士,从事中药学研究,E-mail:2932088632@qq.com
李洪梅,研究员,从事中药药理研究及新药开发,E-mail:lihm2006@sina.cn
黄璐琦,研究员,博士生导师,从事中药资源与分子生药学研究,E-mail:huangluqi01@126.com
收稿日期:2021-01-17,
网络出版日期:2021-04-13,
纸质出版日期:2021-06-20
移动端阅览
席蕴文,康利平,孙建辉等.赤箭和天麻调节免疫功能和肠道菌群作用比较[J].中国实验方剂学杂志,2021,27(12):117-124.
XI Yun-wen,KANG Li-ping,SUN Jian-hui,et al.Comparison of Effect of Chijian and Gastrodiae Rhizoma on Regulating Immune Function and Intestinal Microflora[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(12):117-124.
席蕴文,康利平,孙建辉等.赤箭和天麻调节免疫功能和肠道菌群作用比较[J].中国实验方剂学杂志,2021,27(12):117-124. DOI: 10.13422/j.cnki.syfjx.20211449.
XI Yun-wen,KANG Li-ping,SUN Jian-hui,et al.Comparison of Effect of Chijian and Gastrodiae Rhizoma on Regulating Immune Function and Intestinal Microflora[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(12):117-124. DOI: 10.13422/j.cnki.syfjx.20211449.
目的
2
对赤箭(天麻地上部分)和天麻中腺苷、天麻素、对羟基苯甲醇、对羟基苯甲醛、巴利森苷B和A 的成分含量,以及赤箭水提液和天麻水提液调节免疫功能和肠道菌群作用进行比较,评价赤箭进一步研究和开发的可行性。
方法
2
采用超高效液相色谱法(UPLC)测定6种成分的含量,流动相0.1%甲酸水溶液(A)-乙腈(B)梯度洗脱(0~4 min,0.5%B; 4~5 min,0.5%~2%B;5~10 min,2%~15%B;10~12 min,15%~20%B; 12~15 min,20%~95%B;15~17 min,95%B;17~17.5 min,95%~0.5%B;17.5~20 min,0.5%B),流速0.5 mL·min
-1
,检测波长270 nm。采用环磷酰胺致小鼠免疫功能低下模型,测定廓清指数、校正廓清指数及外周血象,采用体外淋巴细胞转化试验测定B淋巴细胞增殖,基于16S rDNA技术测定肠道菌群并进行生物信息学分析,对赤箭水提液和天麻水提液进行药理活性比较。
结果
2
赤箭粉末、醇提液中6个成分总量分别高于天麻粉末,醇提液,赤箭水提液和天麻水提液中6个成分含量总和相近,且赤箭粉末和天麻粉末中天麻素和对羟基苯甲醇总量均符合2020年版《中华人民共和国药典》的相关要求。与空白组比较,赤箭水提液中剂量(10 g·kg
-1
)组和天麻水提液中、低剂量(10,5 g·kg
-1
)组可明显增加免疫功能低下小鼠的廓清指数(
P
<
0.05);赤箭水提液、天麻水提液高剂量组(20 g·kg
-1
)均可明显增高外周血中红细胞和红细胞压积水平(
P
<
0.05,
P
<
0.01);400 g·L
-1
天麻水提液和100 g·L
-1
赤箭水提液对脂多糖诱导的B淋巴细胞增殖具有促进作用。肠道菌群研究显示,与空白组比较,在门水平上,赤箭水提液和天麻水提液增加拟杆菌门的相对丰度,降低厚壁菌门的相对丰度;在属水平上,赤箭水提液、天麻水提液均可增加
Prevotellaceae
_UCG
-
001和
Ruminococcaceae
_UCG
-
005的相对丰度,降低厌氧棍状菌属,unclassified_
f
_
Erysipelotrichaceae
和
Candidatus
_
Stoquefichus
的相对丰度,这些肠道菌与免疫系统、细胞增殖、调节代谢等相关
。
结论
2
赤箭粉末、醇提液、水提液所测腺苷、天麻素、对羟基苯甲醇、对羟基苯甲醛、巴利森苷B和A总量高于或者接近于天麻相应样品;赤箭水提液与天麻水提液药理活性相似,提示赤箭值得进一步研究与开发。
Objective
2
To compare the contents of adenosine, gastrodin,
p
-hydroxybenzyl alcohol,
p
-hydroxybenzaldehyde, parisinin B and parisinin A in Chijian (the aerial part of
Gastrodia elata
) and Gastrodiae Rhizoma, and compare their effects on immune function and intestinal microflora, evaluating whether it is necessary to study and develop Chijian.
Method
2
The contents of these six constituents were determined by ultra performance liquid chromatography (UPLC), the mobile phase was 0.1% formic acid aqueous solution (A)-acetonitrile (B) for gradient elution (0-4 min, 0.5%B; 4-5 min, 0.5%-2%B; 5-10 min, 2%-15%B; 10-12 min, 15%-20%B; 12-15 min, 20%-95%B; 15-17 min, 95%B; 17-17.5 min, 95%-0.5%B; 17.5-20 min, 0.5%B), the flow rate was 0.5 mL·min
-1
, the detection wavelength was 270 nm. The difference of pharmacological activity of water extracts of Chijian and Gastrodiae Rhizoma was compared, the clearance index, corrected clearance index and peripheral blood were measured in mice model with low immune function induced by cyclophosphamide, B lymphocyte proliferation was determined by lymphocyte transformation test
in vitro
, intestinal microflora was analyzed by 16S rDNA technology and bioinformatics was conducted.
Result
2
The total contents of these six components in powder and ethanol extract of Chijian were higher than that of Gastrodiae Rhizoma, but the total contents of these six components in their water extract were similar, and the total contents of gastrodin and
p
-hydroxybenzyl alcohol met the requirements of the 2020 edition of
Chinese Pharmacopoeia
. Compared with the blank group, the clearance index of immunocompromised mice was significantly increased in the middle-dose (10 g·kg
-1
) group of Chijian water extract, middle- and low-dose (10, 5 g·kg
-1
) groups of Gastrodiae Rhizoma water extract (
P
<
0.05), the levels of erythrocyte and hematocrit in peripheral blood were significantly increased in the high-dose (20 g·kg
-1
) groups of water extracts of Chijian and Gastrodiae Rhizoma (
P
<
0.05,
P
<
0.01), water extract of Gastrodiae Rhizoma with concentration of 400 g·L
-1
and the water extract of Chijian with the concentration of 100 g·L
-1
could promote the proliferation of B lymphocytes induced by lipopolysaccharide. Studies on intestinal microflora showed that compared with the blank group, at the phylum level, the water extracts of Chijian and Gastrodiae Rhizoma increased the relative abundance of Bacteroidetes and decreased the relative abundance of Firmicutes, at the genus level, they increased the relative abundance of
Prevotellaceae
_UCG-001 and
Ruminococcaceae
_UCG-005, and decreased the relative abundance of
Anaerotruncus
, unclassified_
f
_
Erysipelotrichaceae
and
Candidatus
_
Stoquefichus
.
These intestinal bacteria were related to the immune system, cell proliferation, and metabolism regulation.
Conclusion
2
The total contents of 6 components in the powder, the ethanol and the water extracts of Chijian are higher than or close to those of the corresponding samples of Gastrodiae Rhizoma, the pharmacological activity of Chijian water extract is similar to that of Gastrodiae Rhizoma water extract, indicating that Chijian is worthy of further research and development.
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