辽宁中医药大学 药学院,辽宁 大连 116600
凌嘉阳,在读硕士,从事中药化学研究,E-mail:913661241@qq.com
才谦,博士,教授,博士生导师,从事中药化学研究,Tel:0411-85890125,E-mail:caiqianmail@sina.com.
收稿:2023-04-18,
网络出版:2023-07-06,
纸质出版:2024-04-05
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凌嘉阳,柏阳,曲扬等.基于药效学和代谢组学考察北苍术不同炮制品对脾虚大鼠的影响[J].中国实验方剂学杂志,2024,30(07):179-188.
LING Jiayang,BAI Yang,QU Yang,et al.Investigation of Effect of Different Processed Products of Atractylodes chinensis Rhizoma on Rats with Spleen Deficiency Based on Pharmacodynamics and Metabolomics[J].Chinese Journal of Experimental Traditional Medical Formulae,2024,30(07):179-188.
凌嘉阳,柏阳,曲扬等.基于药效学和代谢组学考察北苍术不同炮制品对脾虚大鼠的影响[J].中国实验方剂学杂志,2024,30(07):179-188. DOI: 10.13422/j.cnki.syfjx.20231262.
LING Jiayang,BAI Yang,QU Yang,et al.Investigation of Effect of Different Processed Products of Atractylodes chinensis Rhizoma on Rats with Spleen Deficiency Based on Pharmacodynamics and Metabolomics[J].Chinese Journal of Experimental Traditional Medical Formulae,2024,30(07):179-188. DOI: 10.13422/j.cnki.syfjx.20231262.
目的
2
通过北苍术生品、麸炒品和米泔水制品对脾虚大鼠药效学和代谢组学研究,明确北苍术不同炮制品的药效和作用机制的差异。
方法
2
将60只雄性SD大鼠随机分为空白组、模型组、生苍术组、麸炒苍术组、米泔水制苍术组和参苓白术散组,每组10只,采用疲劳过度加饮食不节法造成大鼠脾虚模型。除空白组和模型组外,上述给药组分别给予生苍术(3.75 g·kg
-1
)、麸炒苍术(3.75 g·kg
-1
)、米泔水制苍术(3.75 g·kg
-1
)和参苓白术散(6.7 g·kg
-1
)混悬液,考察各组大鼠的免疫器官系数,采用酶联免疫吸附测定法(ELISA)检测各组大鼠血浆中与消化道炎症有关指标白细胞介素-6(IL-6)、肿瘤坏死因子-
α
(TNF-
α
)、免疫球蛋白(Ig)G,与消化系统相关的指标淀粉酶(AMS)、胃动素(MTL)、胃泌素(GAS),与燥性相关的指标Na
+
-K
+
-三磷酸腺苷(ATP)酶、大鼠肾组织中水通道蛋白2(AQP2)、大鼠结肠组织中水通道蛋白3(AQP3)与水通道蛋白8(AQP8)的水平。采用超高效液相色谱-四极杆-飞行时间串联质谱法(UPLC-Q-TOF-MS)结合正交偏最小二乘法-判别分析(OPLS-DA)寻找脾虚大鼠血浆样品中的生物标志物[
P
<
0.05和变量重要性投影(VIP)值
>
1],比较3种炮制品对脾虚生物标志物的不同调节作用,并通过京都基因与基因组百科全书(KEGG)数据库进行代谢通路分析。
结果
2
与空白组比较,模型组大鼠胸腺指数和脾脏指数均明显降低(
P
<
0.05),IL-6、TNF-
α
、IgG与AQP2含量明显升高(
P
<
0.05),AMS、GAS、MTL、AQP3、AQP8与Na
+
-K
+
-ATP酶含量明显降低(
P
<
0.05)。与模型组比较,生苍术、麸炒苍术和米泔水制苍术均能提高胸腺指数和脾脏指数(
P
<
0.05),降低IL-6、TNF-
α
、IgG和AQP2(
P
<
0.05),不同程度地提高AMS、GAS、MTL、AQP3、AQP8和Na
+
-K
+
-ATP酶水平。与空白组比较,模型组共筛选出176个差异代谢物,生苍术、麸炒苍术和米泔水制苍术分别回调了其中的75、72、84个生物标志物(
P
<
0.05,
P
<
0.01)。生苍术主要影响了甘氨酸、丝氨酸和苏氨酸代谢。麸炒苍术主要影响了丙氨酸、天冬氨酸和谷氨酸代谢,
D
-精氨酸和
D
-鸟氨酸代谢。米泔水制苍术主要影响了甘氨酸、丝氨酸和苏氨酸代谢,丙氨酸、天冬氨酸和谷氨酸代谢,柠檬酸循环,硫胺素代谢,
D
-精氨酸和
D
-鸟氨酸代谢。
结论
2
生苍术、麸炒苍术和米泔水制苍术均有较好的健脾作用,其中麸炒苍术和米泔水制苍术的作用更强。同时,生苍术燥性最强,麸炒苍术其次,米泔水制苍术燥性最弱。3种苍术炮制品能够显著回调脾虚大鼠代谢异常,其机制可能与调节机体甘氨酸、丝氨酸和苏氨酸代谢,丙氨酸、天冬氨酸和谷氨酸代谢等氨基酸代谢有关。
Objective
2
To clarify the differences in the efficacy and mechanism of different processed products of
Atractylodes chinensis
rhizoma by the pharmacodynamics and metabolomics studies of raw, bran-fried and rice water-processed products on rats with spleen deficiency.
Method
2
Sixty male SD rats were randomly divided into blank group, model group, raw product group(3.75 g·kg
-1
), bran-fried product group(3.75 g·kg
-1
), rice water-processed product group(3.75 g·kg
-1
) and Shenling Baizhusan group(6.7 g·kg
-1
), with 10 rats in each group. The method of excessive fatigue+improper diet was used to establish a spleen deficiency model in rats. After the end of modeling, except for the blank and model groups, each dosing group was given the corresponding drug suspension, the immune organ coefficients of each group of rats were examined, the levels of interleukin-6(IL-6), tumor necrosis factor-
α
(TNF-
α
), immunoglobulin G(IgG), amylase(AMS), motilin(MTL), gastrin(GAS), Na
+
-K
+
-adenosine triphosphatase(ATPase), aquaporin 2(AQP2), AQP3 and AQP8 in rats were measured by enzyme-linked immunosorbent assay(ELISA). Ultra high performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS) combined with orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to search for biomarkers in the plasma samples of spleen-deficient rats by using two criteria[
P
<
0.05 and variable importance in the projection(VIP) value
>
1], and to compare the different modulatory effects of the three decoction pieces on the splenic-deficient biomarkers, and metabolic pathway analysis was conducted through the Kyoto Encyclopedia of Genes and Genomes(KEGG) database.
Result
2
Compared with the blank group, the thymus index and spleen index of rats in the model group were significantly decreased(
P
<
0.05), the levels of IL-6, TNF-
α
, IgG and AQP2 were significantly increased(
P
<
0.05), the levels of AMS, GAS, MTL, AQP3, AQP8 and Na
+
-K
+
-ATPase were significantly decreased(
P
<
0.05). Compared with the model group, raw products, bran-fried products and rice water-processed products all increased thymus index and spleen index(
P
<
0.05), decreased IL-6, TNF-
α
, IgG and AQP2 levels(
P
<
0.05), and increased AMS, GAS, MTL, AQP3, AQP8 and Na
+
-K
+
-ATPase levels to different degrees. A total of 176 differential metabolites were screened in the model group compared with the blank group, of which 75, 72 and 84 biomarkers were called back by the raw products, bran-fried products and rice water-processed products, respectively(
P
<
0.05,
P
<
0.01). Raw products of
A. chinensis
rhizoma mainly affected glycine, serine and threonine metabolism. Bran-fried products mainly affected alanine, aspartate and glutamate metabolism,
D
-arginine and
D
-ornithine metabolism. Rice water-processed products mainly affected glycine, serine and threonine metabolism, alanine, aspartate and glutamate metabolism, citrate cycle, thiamine metabolism,
D
-arginine and
D
-ornithine metabolism.
Conclusion
2
Raw products, bran-fried products and rice water-processed products of
A. chinensis
rhizoma all have good spleen strengthening effects, among which the effects of bran-fried products and rice water-processed products were stronger. Meanwhile, raw products has the strongest dryness, followed by bran-fried products, and the weakest dryness of rice water-processed products. The three decoction pieces are able to significantly modulate metabolic abnormalities in spleen-deficient rats, and the mechanism may be related to amino acid metabolism such as glycine, serine and threonine metabolism as well as alanine, aspartate and glutamate metabolism.
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