Effects of tetramethylpyrazine on expression levels of TNF-α, IL-17 and VEGF in skin lesions of psoriatic mice

doi:10.11958/20221433

Abstract

Abstract:

Objective To investigate the therapeutic effect of tetramethylpyrazine (TMP) on expression levels of tumor necrosis factor-α (TNF-α), interleukin-17 (IL-17) and vascular endothelial growth factor (VEGF) in skin lesions of psoriatic mice and its possible mechanism. Methods Thirty BALB/c mice, half male and half female, were randomly divided into the normal group, the model group and the low, medium and high TMP groups after shaving hair in back area of about 2 cm×4 cm, with 6 mice in each group. Mice in the normal group was smeared with vaseline 62.5 mg/d. Mice in the model group and the TMP group were smeared with imiquimod cream 62.5 mg/d for continuous 7 days. From the 4th day, the low, medium and high TMP groups were respectively injected with ligustrazine hydrochloride 20 mg/kg, 40 mg/kg and 100 mg/kg, and the normal group and the model group were injected with the same amount of normal saline once a day for 9 consecutive days. Changes of skin lesions were observed in each group. After the intervention, back skin lesions of mice were taken. The expression levels of TNF-α, IL-17 and VEGF mRNA in skin lesions were detected by qRT-PCR. The pathological changes of skin lesions and TNF-α, IL-17 and VEGF protein expression levels in skin lesions of each group were detected by HE staining and immunohistochemical staining. Results Mice in the model group showed typical psoriasis-like pathological changes, while the psoriasis-like pathological changes of skin lesions of the TMP group were successively lighter than those of the model group. Compared with the normal group, the expression levels of TNF-α, IL-17 and VEGF mRNA were significantly higher in the model group (all P<0.05). Compared with the model group and the low TMP group, the expression levels of TNF-α mRNA decreased significantly in the medium TMP group and the high TMP group (P<0.05). Compared with the model group, expression levels of IL-17 mRNA were decreased gradually in the three TMP groups (P<0.05). Compared with the model group, the low TMP group and the medium TMP group, the expression level of VEGF mRNA was decreased significantly in the high TMP group (P<0.05). Immunohistochemical results showed that TNF-α, IL-17 and VEGF were mainly expressed in cytoplasm of epidermal cells, and expression levels of above proteins were significantly higher in the model group than those of the normal group (P<0.05). Compared with the model group, expression levels of IL-17 and VEGF were significantly decreased in the low, medium and high TMP groups, and TNF-α was significantly decreased in the medium and high TMP groups (P<0.05). Compared with the low TMP group, expression levels of TNF-α, IL-17 and VEGF were significantly decreased in the high TMP group and IL-17 was also significantly decreased in the medium TMP group (P<0.05). Compared with the medium TMP group, the expression level of VEGF decreased significantly in the high TMP group (P<0.05). Conclusion Tetramethylpyrazine can improve the inflammatory manifestations of psoriasis-like skin lesions in model mice by reducing expression levels of TNF-α, IL-17 and VEGF in psoriasis-like skin lesions of mice.

Key words: psoriasis, tumor necrosis factor-alpha, interleukin-17, vascular endothelial growth factors, tetramethylpyrazine

CLC Number:

R758.63

Figures/Tables 7

References 24

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基因名称	引物序列（5'→3'）	产物大小（bp）
β-actin	上游：CAGCTCAGTAACAGTCCGCC	240
	下游：CCACCATGTACCCAGGCATT	240
TNF-α	上游：GTGAGGAGCACGTAGTCGG	112
	下游：ACGGCATGGATCTCAAAGACA	112
IL-17	上游：TCATTGTGGAGGGCAGACAA	128
	下游：TGACCCCTAAGAAACCCCCA	128
VEGF	上游：AGGAAGGGTAAGCCACTCAC	89
	下游：CCAGGAGGACCTTGTGTGAT	89

基因名称	引物序列（5'→3'）	产物大小（bp）
β-actin	上游：CAGCTCAGTAACAGTCCGCC	240
	下游：CCACCATGTACCCAGGCATT	240
TNF-α	上游：GTGAGGAGCACGTAGTCGG	112
	下游：ACGGCATGGATCTCAAAGACA	112
IL-17	上游：TCATTGTGGAGGGCAGACAA	128
	下游：TGACCCCTAAGAAACCCCCA	128
VEGF	上游：AGGAAGGGTAAGCCACTCAC	89
	下游：CCAGGAGGACCTTGTGTGAT	89

组别	TNF-α	IL-17	VEGF
正常组	-0.039 9±0.180 3	-0.050 6±0.209 7	-0.026 6±0.151 8
模型组	1.037 9±0.292 0^a	0.666 4±0.235 0^a	1.278 9±0.254 7^a
低TMP组	0.852 6±0.159 9^a	-0.280 4±0.074 6^ab	1.137 3±0.165 9^a
中TMP组	0.428 8±0.152 9^abc	-0.547 7±0.143 4^abc	0.801 0±0.139 8^a
高TMP组	0.367 7±0.105 0^abc	-0.778 9±0.158 9^abcd	0.327 0±0.058 7^abcd
F	25.437^＊＊	51.276^＊＊	54.334^＊＊

组别	TNF-α	IL-17	VEGF
正常组	-0.039 9±0.180 3	-0.050 6±0.209 7	-0.026 6±0.151 8
模型组	1.037 9±0.292 0^a	0.666 4±0.235 0^a	1.278 9±0.254 7^a
低TMP组	0.852 6±0.159 9^a	-0.280 4±0.074 6^ab	1.137 3±0.165 9^a
中TMP组	0.428 8±0.152 9^abc	-0.547 7±0.143 4^abc	0.801 0±0.139 8^a
高TMP组	0.367 7±0.105 0^abc	-0.778 9±0.158 9^abcd	0.327 0±0.058 7^abcd
F	25.437^＊＊	51.276^＊＊	54.334^＊＊

组别	TNF-α	IL-17	VEGF
正常组	0.004±0.002	0.002±0.001	0.001±0.001
模型组	0.019±0.003^a	0.022±0.001^a	0.021±0.003^a
低TMP组	0.017±0.003^a	0.015±0.001^ab	0.019±0.001^ab
中TMP组	0.014±0.001^ab	0.009±0.001^abc	0.012±0.001^ab
高TMP组	0.013±0.001^abc	0.008±0.001^abc	0.008±0.001^abcd
F	98.959^＊＊	718.474^＊＊	367.192^＊＊