Nicotine enhanced the inducible expression of HSP70 under heat stress through N1 receptor in RAW264.7 macrophages

Abstract

Abstract:

AIM: To explore whether low doses of nicotine enhances the HSF1 nuclear translocation and expression of HSP70 in the RAW264.7 macrophages treated under lower heat stress (<40 ℃), and to uncover its receptor mechanism. METHODS:After stimulating with nicotine at doses of 1 mmol/L and its antagonists prior to nicotine treatment at 37 ℃ or 39 ℃, the expression of HSP70 in RAW264.7 macrophages was detected by Western-blot, translocation of HSF1 was detected by immunofluorescence and the combination of HSF1and heat shock element (HSE) was detected by EMSA, respectively. RESULTS:Western-blot showed that nicotine (1 mmol/L) alone could not induce HSPs expression, but was able to enhance the HSP70 expression at 39 ℃ in RAW246.7. Immunofluorescence showed nicotine (1 mmol/L) was able to induce translocation of HSF1 from cytoplasm to nucleus, which was blocked by the pre-treating with hexamethonium apparently; EMSA showed that nicotine (1 mmol/L) could promote the binding of HSF1 to HSE at 37℃. Furthermore, nicotine (1 mmol/L) induced HSF1 binding to HSE greatly at 39℃. CONCLUSION: Nicotine (1 mmol/L) alone can induce HSF1 nuclear translocation and its DNA-binding activity to HSE, but not the expression of HSP70; nicotine (1 mmol/L) can enhance the expression of HSP70 under mild heat stress(39℃) through N1-receptor in the RAW264.7 macrophages.

Key words: nicotine, HSF1, HSP70, N1 receptor

CLC Number:

R965.2

CHEN Chongfeng, YANG Huaicai, TU Zizhi. Nicotine enhanced the inducible expression of HSP70 under heat stress through N1 receptor in RAW264.7 macrophages[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(2): 124-131.

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