The effect of Xijiao Dihuang combined prescription on T cell differentiation induced by platelet antigen loaded DC in ITP patients

doi:10.11958/20220771

Abstract

Abstract:

Objective To observe the effect of drug-containing serum of Xijiao Dihuang combined prescription (XJDHCP) on the proliferation and differentiation of dendritic cell (DC)-activated T cells and its mechanism on immune thrombocytopenia (ITP). Methods Ten male SD rats were randomly divided into the drug-containing serum group and the blank serum group, with 5 rats in each group. Rats were gavaged with XJDHCF and distilled water for 3 days. Blood-extraction from the abdominal aorta was prepared for drug-containing serum and blank serum. Fifteen healthy volunteers and 8 ITP patients with CD4⁺ T cells were selected, and the DCs loaded with platelet antigens were co-cultured with CD4⁺ T cells and randomly divided into the normal control group, the model group and the low, medium and high (5%, 10% and 20%) dose groups of XJDHCF. The normal control group consisted of platelet antigen-loaded DCs and CD4⁺ T cells from healthy volunteers, and the model group and the low, medium and high dose groups of XJDHCF were platelet antigen-loaded DCs and CD4⁺ T cells from ITP patients, in which the normal control group and the model group were added with rat blank serum, and the low, medium and high dose groups of XJDHCF were added with 5%, 10% and 20% drug-containing serum. The proliferation of CD4⁺ T cells, the ratio of regulatory T cells (Treg), effector T cells (Teff) and the expression of programmed death receptor-1 (PD-1) on the surface of CD4⁺ T were detected by flow cytometry (FCM). Enzyme-linked immunosorbent assay was used to detect the expression levels of pro-inflammatory factors IL-2, IFN-γ, IL-17 and anti-inflammatory factors TGF-β and IL-10 in the supernatant of each group. Results Compared with the control group, the percentage of CD4⁺ T cell proliferation, the proportion of Teff cells and amounts of pro-inflammatory factors IL-2, IFN-γ and IL-17 were significantly higher in the model group (P<0.05), and the proportion of Treg cells, the amount of PD-1 expression on the surface of CD4⁺ T cells and amounts of anti-inflammatory factors TGF-β and IL-10 were greatly lower in the model group (P<0.05). Compared with the model group, the percentage of CD4⁺ T cell proliferation and the proportion of Teff cells decreased successively in the low, medium and high dose groups of XJDHCF (P<0.05), and the amount of PD-1 expression on the surface of CD4⁺ T cells and the amounts of TGF-β, an inflammatory factor were increased (P<0.05). The amount of IFN-γ was significantly lower in the high dose group of XJDHCF (P<0.05). Compared with the model group, the amounts of IFN-γ and IL-17 were significantly lower in the middle and high dose groups of XJDHCF (P<0.05). Conclusion XJDHCF can regulate the excessive proliferation and differentiation of CD4⁺ T cells and the secretion of cytokines in ITP patients in vitro, which may be one of the mechanisms of XJDHCF in the treatment of ITP.

Key words: thrombocytopenia, dendritic cells, CD4-positive T-lymphocytes, disease models, animal, Xijiao Dihuang combined prescription

CLC Number:

R558.2

Figures/Tables 6

References 13

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组别	Treg	Teff
对照组	8.82±0.11	67.75±1.20
模型组	3.46±1.11^a	82.54±0.66^a
犀角地黄汤合方低剂量组	4.09±0.13^ab	79.50±0.69^ab
犀角地黄汤合方中剂量组	5.23±0.95^abc	76.57±0.97^abc
犀角地黄汤合方高剂量组	6.47±0.45^abc	72.02±1.70^abcd
F	747.550^＊＊	83.971^＊＊

组别	Treg	Teff
对照组	8.82±0.11	67.75±1.20
模型组	3.46±1.11^a	82.54±0.66^a
犀角地黄汤合方低剂量组	4.09±0.13^ab	79.50±0.69^ab
犀角地黄汤合方中剂量组	5.23±0.95^abc	76.57±0.97^abc
犀角地黄汤合方高剂量组	6.47±0.45^abc	72.02±1.70^abcd
F	747.550^＊＊	83.971^＊＊

组别	IL-2		IL-17	IL-10	TGF-β
对照组	15.77±3.46	150.24±10.71	118.80±3.27	47.22±2.54	609.51±9.39
模型组	137.56±7.19^a	809.56±41.18^a	520.29±9.96^a	16.45±3.70^a	200.84±3.69^a
犀角地黄汤合方低剂量组	51.71±3.30^ab	685.93±7.45^a	515.86±3.51^a	27.76±3.05^ab	213.98±1.91^ab
犀角地黄汤合方中剂量组	27.98±0.51^abc	467.41±21.54^abc	456.48±5.25^abc	31.81±2.43^ab	311.57±3.60^abc
犀角地黄汤合方高剂量组	27.52±1.53^abc	381.29±56.48^bcd	327.08±6.59^abcd	41.54±4.39^bc	481.23±7.04^abcd
F	476.357^＊＊	915.451^＊＊	2 226.488^＊＊	39.683^＊＊	2 810.404^＊＊

组别	IL-2		IL-17	IL-10	TGF-β
对照组	15.77±3.46	150.24±10.71	118.80±3.27	47.22±2.54	609.51±9.39
模型组	137.56±7.19^a	809.56±41.18^a	520.29±9.96^a	16.45±3.70^a	200.84±3.69^a
犀角地黄汤合方低剂量组	51.71±3.30^ab	685.93±7.45^a	515.86±3.51^a	27.76±3.05^ab	213.98±1.91^ab
犀角地黄汤合方中剂量组	27.98±0.51^abc	467.41±21.54^abc	456.48±5.25^abc	31.81±2.43^ab	311.57±3.60^abc
犀角地黄汤合方高剂量组	27.52±1.53^abc	381.29±56.48^bcd	327.08±6.59^abcd	41.54±4.39^bc	481.23±7.04^abcd
F	476.357^＊＊	915.451^＊＊	2 226.488^＊＊	39.683^＊＊	2 810.404^＊＊

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