Transcriptomics-based anti-tuberculous mechanism of traditional Chinese polyherbal preparation NiuBeiXiaoHe intermediates

Abstract

Background: Integrated traditional Chinese medicine and biomedicine is an effective method to treat tuberculosis (TB). In our previous research, traditional Chinese medicine preparation NiuBeiXiaoHe (NBXH) achieved obvious anti-TB effects in animal experiments and clinical practice. However, the action mechanism of NBXH has not been elucidated.

Method: Peripheral blood mononuclear cells (PBMCs) were collected to extract mRNA and differentially expressed (DE) genes were obtained using gene microarray technology. Finally, GEO databases and RT-qPCR were used to verify the results of expression profile.

Result: After MTB infection, most upregulated DE genes in mice were immune-related genes, including cxcl9, camp, cfb, c4b, serpina3g, and ngp. Downregulated DE genes included lrrc74b, sult1d1, cxxc4, and grip2. After treatment with NBXH, especially high-dose NBXH, the abnormal gene expression was significantly corrected. Some DE genes have been confirmed in multiple GEO datasets or in pulmonary TB patients through RT-qPCR.

Conclusion: MTB infection led to extensive changes in host gene expression and mainly caused the host's anti-TB immune responses. The treatment using high-dose NBXH partially repaired the abnormal gene expression, further enhanced the anti-TB immunity included autophagy and NK cell-mediated cytotoxicity, and had a certain inhibitory effect on overactivated immune responses.

Keywords: Chinese traditional medicine; NiuBeiXiaoHe; mechanism of action; transcriptome; tuberculosis.

FIGURE 1

The study flowchart.

FIGURE 2

The DE genes in significantly upregulated and downregulated pathways after MTB infection. (A) Each pathway shown in Table 5, and the significantly upregulated DE genes in each pathway after MTB infection; (B) Each pathway shown in Table 6, and the significantly downregulated DE genes in each pathway after MTB infection. Red represents the upregulated DE genes, green represents unchanged genes, and blue represents downregulated DE genes.

FIGURE 3

Expression levels of genes ifn-γ, cxcl9, cfb, cxxc4, ulk3, camp, and cd302 in the human PBMC samples as determined by RT-qPCR HC (healthy controls): negative IGRA and without abnormality in lung CT; TB (tuberculosis): initial treatment TB patients with positive IGRA or smear or Xpert. The data between the two groups were tested by Student’s t-test. Results of cytokine relative expression were expressed as a scatter chart, in which (A) means ifn-γ (p-value < 0.0001); (B) means cxcl9 (p-value < 0.0001); (C) means cfb (p-value = 0.0003); (D) means cxxc4 (p-value = 0.0493); (E) means ulk3 (p-value = 0.0227); (F) means camp (p-value = 0.0362); (G) means cd302 (p-value = 0.4306).

FIGURE 4

The DE gene Venn diagram between TB model group vs. normal group, JHW group vs. TB model group, and each dose NBXH group vs. TB model group. (A) Downregulated DE genes in the TB model group vs. normal group and upregulated DE genes in JHW group vs. TB model group and each NBXH group vs. TB model group; (B) Upregulated DE genes in the TB model group vs. normal group and downregulated DE genes in JHW group vs. TB model group and each NBXH group vs. TB model group, intersecting parts mean overlapping DE genes.

FIGURE 5

The DE genes in significantly upregulated and downregulated pathways after NBXH high dose group vs. TB model group. (A) The diagram of each pathway shown in Supplementary Table S3, and the significantly upregulated DE genes in each pathway after the high-dose NBXH treatment on mouse TB model; (B) The diagram of each pathway shown in Supplementary Table S4, and the significantly downregulated DE genes in each pathway after the high-dose NBXH treatment on mouse TB model. Red represents the upregulated DE genes, green represents unchanged genes, and blue represents downregulated DE genes.

FIGURE 6

Transcript levels of ifn-γ, cxcl9, cfb, cxxc4, and ulk3 genes in the human PBMCs samples as determined by RT-qPCR.