Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (4): 794-803.DOI: 10.19852/j.cnki.jtcm.20240515.001
• Original articles • Previous Articles Next Articles
LUO Shan1, YANG Fan2, CHEN Yuanchun2, ZHAO Ruoxi2, LIU Haiye1, GAO Fei3, MA Wencan3, GAO Weijuan1,4(), YU Wentao3,4,5()
Received:
2023-01-11
Accepted:
2023-07-16
Online:
2024-08-15
Published:
2024-05-15
Contact:
Prof. YU Wentao, College of Acupuncture and Massage, Hebei University of Chinese Medicine, Shijiazhuang 050200, China, Supported by:
LUO Shan, YANG Fan, CHEN Yuanchun, ZHAO Ruoxi, LIU Haiye, GAO Fei, MA Wencan, GAO Weijuan, YU Wentao. Sanhua Tang (三化汤) protects against ischemic stroke by preventing blood-brain barrier injury: a network pharmacology and in vivo experiments[J]. Journal of Traditional Chinese Medicine, 2024, 44(4): 794-803.
Figure 1 Compound-targets network and Herb-pathway-targets network A: the network had 121 nodes representing the potential active components of SHT and 353 edges, describing the relationship between them. The triangle represents IS, and the square represents the four herbs of SHT. DH (Dahuang), ZS (Zhishi), QH (Qianghuo), and HP (Houpu) stand for herbs' abbreviation. The hexagon represents effective chemical compounds, and the diamond represents targets. B: circles represent compounds, arrows represent pathways, squares represent the top 10 core targets, and triangles represent IS. SHT: Sanhua Tang; IS: ischemic stroke.
Figure 2 Conformations of some key compounds and core targets A: AKT1 with luteolin; B: IL-6 with luteolin; C: TNF with luteolin; D: CASP3 with luteolin; E: AKT1 with naringenin; F: CASP3 with beta-sitosterol. GLN: glutamine; ASN: asparagine; LYS: lysine; ALA: alanine; MET: methionine; LEU: leucine; ARG: arginine; PHE: phenylalanine; TRP: tryptophan; AKT1: v-akt murine thymoma viral oncogene homolog 1; IL-6: Interleukin-6; TNF: tumor necrosis factor; CASP3: cysteinyl aspartate specific proteinase 3.
Neurological dysfunction | ANOVA | Treatment (between columns) | Residual (within columns) | F (DFn, DFd) | P value |
---|---|---|---|---|---|
Neurological score | SS | 924.4 | 447.4 | F (5, 102) = 42.15 | < 0.0001 |
DF | 5 | 102 | |||
MS | 184.9 | 4.387 | |||
Infarct area | SS | 3.165 | 0.6348 | F (5, 23) = 22.93 | < 0.0001 |
DF | 5 | 23 | |||
MS | 0.6329 | 0.02760 | |||
Number of positive cells | SS | 19256 | 5910 | F (5, 30) = 19.55 | < 0.0001 |
DF | 5 | 30 | |||
MS | 3851 | 197.0 |
Table 1 Statistical results of neurological score, infarct area and the number of positive cells between each group after ANOVA
Neurological dysfunction | ANOVA | Treatment (between columns) | Residual (within columns) | F (DFn, DFd) | P value |
---|---|---|---|---|---|
Neurological score | SS | 924.4 | 447.4 | F (5, 102) = 42.15 | < 0.0001 |
DF | 5 | 102 | |||
MS | 184.9 | 4.387 | |||
Infarct area | SS | 3.165 | 0.6348 | F (5, 23) = 22.93 | < 0.0001 |
DF | 5 | 23 | |||
MS | 0.6329 | 0.02760 | |||
Number of positive cells | SS | 19256 | 5910 | F (5, 30) = 19.55 | < 0.0001 |
DF | 5 | 30 | |||
MS | 3851 | 197.0 |
Figure 3 Effect of SHT on neurological dysfunction A: TTC staining of cerebral slices of each group; B: Nissl staining observation of ischemic penumbra of rats in each group under × 400 magnification. B1: the sham group; B2: the M group; B3: the SL group; B4: the SM group; B5: the SH group; B6: the AE group. Microscopic observation showed that the green arrows were normal neurons, the neurons were regularly arranged, the whole blue staining and the Nissl corpuscles were dark blue, thick, and dense granules. The red arrows were degenerative and necrotic neurons, the arrangement of neurons was disordered, Nissl corpuscles decreased or even disappeared, and the whole was in a lightly stained state. M: middle cerebral artery occlusion; AE: aloeemodin; SL: Sanhua Tang at low dosage; SM: medium dosage; SH: high dosage; SHT: Sanhua Tang; TTC: triphenyltetrazolium chloride; MCAO: middle cerebral artery occlusion. M: model group were orally administered with saline 1 mL/100 g per day for 5 d; SL: this group were orally administered Sanhua Tang 0.18 g/mL before stroke induction once daily for 4 d, and continued administration for one day after MCAO surgery. As the same goes SM 0.36 g/mL, SH group 0.72 g/mL, and AE 5 mg/mL.
Figure 4 Effect of SHT on inflammatory damage of BBB A: comparison of the protein expression levels of VEGFA in rats. [Mean (SD), n = 4]. B: comparison of BBB ultrastructure under TEM in each group. Each group had two views, B1-6 at low power (× 3000) and B7-12 at high power (× 8000); B1, B7: the sham group; B2, B8: the M group; B3, B9: the AE group; B4, B10: the SL group; B5, B11: the SM group; B6, B12: the SH group. M: middle cerebral artery occlusion; AE: aloeemodin; SL: Sanhua Tang at low dosage; SM: medium dosage; SH: high dosage; SHT: Sanhua Tang; MCAO: middle cerebral artery occlusion; BBB: blood-brain barrier; TEM: transmission electron microscope. M: model group were orally administered with saline 1 mL/100 g per day for 5 d; SL: this group were orally administered Sanhua Tang 0.18 g/mL before stroke induction once daily for 4 d, and continued administration for one day after MCAO surgery. As the same goes SM 0.36 g/mL, SH group 0.72 g/mL, and AE 5 mg/mL.
Inflammatory damage of BBB | ANOVA | Treatment (between columns) | Residual (within columns) | F (DFn, DFd) | P value |
---|---|---|---|---|---|
Levels of IL-6 | SS | 966.0 | 954.9 | F (5, 30) = 6.070 | 0.0005 |
DF | 5 | 30 | |||
MS | 193.2 | 31.83 | |||
Levels of IL-10 | SS | 109.4 | 172.7 | F (5, 30) = 3.802 | 0.0087 |
DF | 5 | 30 | |||
MS | 21.89 | 5.757 | |||
Levels of TNF-α | SS | 99.52 | 171.7 | F (5, 30) = 3.478 | 0.0135 |
DF | 5 | 30 | |||
MS | 19.90 | 5.723 | |||
Levels of VEGFA | SS | 3.438 | 0.6922 | F (5, 18) = 17.88 | < 0.0001 |
DF | 5 | 18 | |||
MS | 0.6876 | 0.03846 |
Table 2 Statistical results of inflammatory factors and VEGFA protein expression levels between each group after ANOVA
Inflammatory damage of BBB | ANOVA | Treatment (between columns) | Residual (within columns) | F (DFn, DFd) | P value |
---|---|---|---|---|---|
Levels of IL-6 | SS | 966.0 | 954.9 | F (5, 30) = 6.070 | 0.0005 |
DF | 5 | 30 | |||
MS | 193.2 | 31.83 | |||
Levels of IL-10 | SS | 109.4 | 172.7 | F (5, 30) = 3.802 | 0.0087 |
DF | 5 | 30 | |||
MS | 21.89 | 5.757 | |||
Levels of TNF-α | SS | 99.52 | 171.7 | F (5, 30) = 3.478 | 0.0135 |
DF | 5 | 30 | |||
MS | 19.90 | 5.723 | |||
Levels of VEGFA | SS | 3.438 | 0.6922 | F (5, 18) = 17.88 | < 0.0001 |
DF | 5 | 18 | |||
MS | 0.6876 | 0.03846 |
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