Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (4): 722-733.DOI: 10.19852/j.cnki.jtcm.20231226.001
• Original articles • Previous Articles Next Articles
YAN Jing1, FENG Huimin1, QIU Fang1, WANG Haijun1, YIN Luyun1, JIN Xiaofei1(), ZHAO Jiyu2, WANG Hongyang1, YAN Xiaoqin3
Received:
2023-05-23
Accepted:
2023-09-12
Online:
2024-08-15
Published:
2023-12-26
Contact:
JIN Xiaofei, Second Clinical College of Shanxi University of Traditional Chinese Medicine, Jinzhong 030619, China. Supported by:
YAN Jing, FENG Huimin, QIU Fang, WANG Haijun, YIN Luyun, JIN Xiaofei, ZHAO Jiyu, WANG Hongyang, YAN Xiaoqin. Effect on serum metabolomics of rats with premature ovarian insufficiency by Zhibian (BL54) through Shuidao (ST28) acupuncture[J]. Journal of Traditional Chinese Medicine, 2024, 44(4): 722-733.
Group | n | Before modeling | After modeling | Post intervention |
---|---|---|---|---|
Control | 12 | 245±7 | 266±11 | 316±15 |
POI | 12 | 244±10 | 253±11a | 286±13b |
SA+POI | 12 | 244±7 | 255±12a | 288±13 |
ZS+POI | 12 | 244±9 | 255±14a | 301±17c |
Table 1 Changes in body weight of rats (g, $\bar{x}±s$)
Group | n | Before modeling | After modeling | Post intervention |
---|---|---|---|---|
Control | 12 | 245±7 | 266±11 | 316±15 |
POI | 12 | 244±10 | 253±11a | 286±13b |
SA+POI | 12 | 244±7 | 255±12a | 288±13 |
ZS+POI | 12 | 244±9 | 255±14a | 301±17c |
Group | n | After modeling | Post-intervention | |||||
---|---|---|---|---|---|---|---|---|
FSH (IU/L) | LH (mIU/mL) | E2 (pmol/L) | FSH (IU/L) | LH (mIU/mL) | E2 (pmol/L) | |||
Control | 12 | 3.1±0.9 | 22.7±6.7 | 39.3±9.8 | 4.0±1.1 | 20.9±5.0 | 43.7±6.1 | |
POI | 12 | 6.0±1.3a | 36.7±5.4a | 26.0±8.7a | 7.4±1.4a | 36.3±4.2a | 28.0±6.0a | |
SA+POI | 12 | 5.8±0.9a | 38.8±5.1a | 29.6±6.5a | 7.3±0.9 | 35.2±4.4 | 30.6±6.1 | |
ZS+POI | 12 | 6.3±0.9a | 36.8±6.2a | 27.1±8.6a | 6.1±0.9b | 30.3±4.4b | 38.0±7.0b |
Table 2 Changes in serum hormone levels ($\bar{x}±s$)
Group | n | After modeling | Post-intervention | |||||
---|---|---|---|---|---|---|---|---|
FSH (IU/L) | LH (mIU/mL) | E2 (pmol/L) | FSH (IU/L) | LH (mIU/mL) | E2 (pmol/L) | |||
Control | 12 | 3.1±0.9 | 22.7±6.7 | 39.3±9.8 | 4.0±1.1 | 20.9±5.0 | 43.7±6.1 | |
POI | 12 | 6.0±1.3a | 36.7±5.4a | 26.0±8.7a | 7.4±1.4a | 36.3±4.2a | 28.0±6.0a | |
SA+POI | 12 | 5.8±0.9a | 38.8±5.1a | 29.6±6.5a | 7.3±0.9 | 35.2±4.4 | 30.6±6.1 | |
ZS+POI | 12 | 6.3±0.9a | 36.8±6.2a | 27.1±8.6a | 6.1±0.9b | 30.3±4.4b | 38.0±7.0b |
Figure 1 ovarian tissue morphology A: gross ovarian tissue specimen in control group; B: gross ovarian tissue specimen in POI group; C: gross ovarian tissue specimen in SA + POI group; D: gross ovarian tissue specimen in ZS + POI group; E: pathomorphological change ovarian tissue in control group; F: pathomorphological change ovarian tissue in POI group; G: pathomorphological change ovarian tissue in SA + POI group; H: pathomorphological change of ovarian tissue in ZS + POI group (HE, × 200). Control: normal feeding, without any treatment. POI: POI model was induced by intraperitoneal injection of CTX (2 mg/mL) for 15 d; SA+POI: sham acupuncture intervention in CTX-induced POI rats for 4 weeks; ZS + POI: Zhibian (BL54) through Shuidao (ST28) acupuncture intervention in CTX-induced POI rats for 4 weeks. PF: primary follicle; EA: early antral follicle; AF: atresia follicle; HE: hematoxylin-eosin staining; POI: premature ovarian insufficiency; CTX: cyclophosphamide; SA: sham acupuncture; ZS: Zhibian (BL54) through Shuidao (ST28) acupuncture.
Group | n | Left ovary | Right ovary | |||
---|---|---|---|---|---|---|
Weight (mg) | Coefficient (mg/g) | Weight (mg) | Coefficient (mg/g) | |||
Control | 12 | 73.333±6.513 | 0.233±0.023 | 71.667±12.673 | 0.227±0.037 | |
POI | 12 | 44.167±12.401a | 0.153±0.039a | 45.000±7.977a | 0.157±0.026a | |
SA+POI | 12 | 45.833±9.962 | 0.159±0.033 | 43.333±10.731 | 0.150±0.032 | |
ZS+POI | 12 | 56.667±6.513b | 0.188±0.020b | 58.333±11.146b | 0.194±0.037b |
Table 3 Changes in ovarian weight and coefficient ($\bar{x}±s$)
Group | n | Left ovary | Right ovary | |||
---|---|---|---|---|---|---|
Weight (mg) | Coefficient (mg/g) | Weight (mg) | Coefficient (mg/g) | |||
Control | 12 | 73.333±6.513 | 0.233±0.023 | 71.667±12.673 | 0.227±0.037 | |
POI | 12 | 44.167±12.401a | 0.153±0.039a | 45.000±7.977a | 0.157±0.026a | |
SA+POI | 12 | 45.833±9.962 | 0.159±0.033 | 43.333±10.731 | 0.150±0.032 | |
ZS+POI | 12 | 56.667±6.513b | 0.188±0.020b | 58.333±11.146b | 0.194±0.037b |
Figure 2 OPLS-DA score plots and model replacement test plots of rat serum samples A: OPLS-DA score plot and model replacement test plot of the control group vs the POI group in positive ion mode; B: OPLS-DA score plot and model replacement test plot of the control group vs the POI group in negative ion mode; C: OPLS-DA score plot and model replacement test plot of the POI group vs the ZS + POI group in positive ion mode; D: OPLS-DA score plot and model replacement test plot of the POI group vs the ZS+POI group in negative ion mode; A1, B1, C1, D1: OPLS-DA score plot; A2, B2, C2, D2: model replacement test plot. In the OPLS-DA score plots, the x-axis (t1) represents the prediction component (intergroup difference component), the y-axis (t2) represents the orthogonal component (intragroup difference component), and the percentage of the horizontal y-axis represents the proportion of this component in the total variance. In the model replacement test plots, the x-axis represents the correlation between the permutation group and the original model group, the y-axis represents the value of R2Y or Q2Y (where R2γ and Q2γ with 1 in the x-axis are the values of the original model), the blue point and the red point represent the R2γ and Q2γ of the replaced model, respectively, and the imaginary line is the fitted regression line. Control: normal feeding, without any treatment; POI: POI model was induced by intraperitoneal injection of CTX (2 mg/mL) for 15 d; ZS+POI: Zhibian (BL54) through Shuidao (ST28) acupuncture intervention in CTX-induced POI rats for 4 weeks. ESI: electron spray ionization; RMSEE: root mean square error, pre: number of predicted components, ort: number of orthogonal components; OPLS-DA: orthogonal partial least squares-discrimination analysis; POI: premature ovarian insufficiency; CTX: cyclophosphamide; ZS: Zhibian (BL54) through Shuidao (ST28) acupuncture.
Figure 3 Screening of metabolic differentials and KEGG pathway enrichment maps in serum of POI model intervened by acupuncture intervention A: volcano map in positive ion mode, B: volcano map in negative ion mode, C: KEGG pathway enrichment map in positive ion mode, D: KEGG pathway enrichment map in negative ion mode. In the volcano map, the x-axis represents the difference multiple, the y-axis represents the P-value, and the scatter size represents the VIP value. Blue dots indicate down-regulation, red dots indicate up-regulation, and gray dots indicate no significant difference. In the KEGG pathway enrichment map, the x-axis is the ratio of the number of differential metabolites to the total number of metabolites, and the y-axis is the name of the pathway. The larger the point, the more the number of metabolites enriched, and the redder the color of the point, the more significant the enrichment pathway. KEGG: Kyoto Encyclopedia of Genes and Genomes; POI: premature ovarian insufficiency.
Name | Formula | Mass-to-charge ratio (m/z) | RT (min) | Control vs POI | POI vs ZS+POI | |||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
FC | P value | VIP | Trend | FC | P value | VIP | Trend | |||||
Divanillyltetrahydrofuran ferulate | C30H32O8 | 1099.4293 | 11.4854 | 0.2409 | 0.0001 | 2.1464 | ↓a | 4.4102 | 0.0000 | 2.2110 | ↑c | |
Rolitetracycline | C27H33N3O8 | 1099.4546 | 13.5905 | 0.1087 | 0.0011 | 2.0491 | ↓b | 6.5169 | 0.0000 | 2.1883 | ↑c | |
Trans-ferulic acid | C10H10O4 | 239.0564 | 2.9167 | 0.6022 | 0.0006 | 1.9304 | ↓a | 1.5964 | 0.0027 | 1.8209 | ↑b | |
Tryptamine | C10H12N2 | 159.0912 | 2.4098 | 0.7189 | 0.0023 | 1.8795 | ↓b | 4.2828 | 0.0000 | 2.2682 | ↑c | |
Neuraminic acid | C9H17NO8 | 288.0653 | 2.0602 | 0.4761 | 0.0038 | 1.8148 | ↓b | 1.6599 | 0.0095 | 1.6740 | ↑b | |
Eupatilin | C18H16O7 | 343.0840 | 4.2009 | 0.5563 | 0.0030 | 1.7867 | ↓b | 2.0420 | 0.0015 | 1.8760 | ↑b |
Table 4 Effect of "Zhibian (BL54) through Shuidao (ST28)" acupuncture on potential biomarkers in the serum of rats with POI
Name | Formula | Mass-to-charge ratio (m/z) | RT (min) | Control vs POI | POI vs ZS+POI | |||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
FC | P value | VIP | Trend | FC | P value | VIP | Trend | |||||
Divanillyltetrahydrofuran ferulate | C30H32O8 | 1099.4293 | 11.4854 | 0.2409 | 0.0001 | 2.1464 | ↓a | 4.4102 | 0.0000 | 2.2110 | ↑c | |
Rolitetracycline | C27H33N3O8 | 1099.4546 | 13.5905 | 0.1087 | 0.0011 | 2.0491 | ↓b | 6.5169 | 0.0000 | 2.1883 | ↑c | |
Trans-ferulic acid | C10H10O4 | 239.0564 | 2.9167 | 0.6022 | 0.0006 | 1.9304 | ↓a | 1.5964 | 0.0027 | 1.8209 | ↑b | |
Tryptamine | C10H12N2 | 159.0912 | 2.4098 | 0.7189 | 0.0023 | 1.8795 | ↓b | 4.2828 | 0.0000 | 2.2682 | ↑c | |
Neuraminic acid | C9H17NO8 | 288.0653 | 2.0602 | 0.4761 | 0.0038 | 1.8148 | ↓b | 1.6599 | 0.0095 | 1.6740 | ↑b | |
Eupatilin | C18H16O7 | 343.0840 | 4.2009 | 0.5563 | 0.0030 | 1.7867 | ↓b | 2.0420 | 0.0015 | 1.8760 | ↑b |
Figure 4 ROC curve and box plot of acupuncture on potential biomarkers in serum of rats with POI A: divanillyltetrahydrofuran ferulate; B: trans-ferulic acid; C: tryptamine; D: neuraminic acid. Each plot represents the ROC curve and metabolic box plot of the same metabolite in the Control vs POI group (above) and ZS + POI vs POI group (below). A1, B1, C1, D1: ROC curve plot in the Control vs POI group; A2, B2, C2, D2: box plot in the Control vs POI group; A3, B3, C3, D3: ROC curve plot in the ZS+POI vs POI group; A4, B4, C4, D4: box plot in the ZS + POI vs POI group. In the ROC curve plot, the area surrounded by the blue dot solid line and the straight line (x = 1, y = 0) is AUC, the closer the AUC value is to 1, the higher the accuracy of the prediction. In the box plot, the content distribution of metabolites in each group was shown. Control: normal feeding, without any treatment; POI: POI model was induced by intraperitoneal injection of CTX (2 mg/mL) for 15 d; SA + POI: sham acupuncture intervention in CTX-induced POI rats for 4 weeks; ZS + POI: Zhibian (BL54) through Shuidao (ST28) acupuncture intervention in CTX-induced POI rats for 4 weeks. ROC: receiver operating characteristic curve; POI: premature ovarian insufficiency; ZS: Zhibian (BL54) through Shuidao (ST28) acupuncture; AUC: area under roc; CTX: cyclophosphamide.
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