ABSTRACT
Both cholinergic dysfunction and protein citrullination are the hallmarks of rheumatoid arthritis (RA), but the relationship between the two phenomena remains unclear. We explored whether and how cholinergic dysfunction accelerates protein citrullination and consequently drives the development of RA. Cholinergic function and protein citrullination levels in patients with RA and collagen-induced arthritis (CIA) mice were collected. In both neuron-macrophage coculture system and CIA mice, the effect of cholinergic dysfunction on protein citrullination and expression of peptidylarginine deiminases (PADs) was assessed by immunofluorescence. The key transcription factors for PAD4 expression were predicted and validated. Cholinergic dysfunction in the patients with RA and CIA mice negatively correlated with the degree of protein citrullination in synovial tissues. The cholinergic or alpha7 nicotinic acetylcholine receptor (α7nAChR) deactivation and activation resulted in the promotion and reduction of protein citrullination in vitro and in vivo, respectively. Especially, the activation deficiency of α7nAChR induced the earlier onset and aggravation of CIA. Furthermore, deactivation of α7nAChR increased the expression of PAD4 and specificity protein-3 (SP3) in vitro and in vivo. Our results suggest that cholinergic dysfunction-induced deficient α7nAChR activation, which induces the expression of SP3 and its downstream molecule PAD4, accelerating protein citrullination and the development of RA.
ABSTRACT
AIM: To established a method for determinating emodin and berberine hydrochloride in Wangshi baochi (Pills Radix et Rhizoma Rhei,Rhizoma Coptidis,etc.) by HPLC. METHODS : The separation was performed on Lichrospher C_ 18 column,using methanol-0.1% H_3PO_4(90∶10) and acetonitrile-0.05mol?L -1 KH_2PO_4 (35∶65) as mobile phase,respectively. The detection wavelength was at 290nm and 345nm,respectively. The flow rate was at 1.0mL?min -1 ,the column temperature was room temperature. RESULTS : The linear range of emodin was in the range of 0.1966～0.9830 ?g,r =0.9997, the average recovery was 98.81% and RSD was 1.41%,respectively. The linear range of berberine hydrochloride was 0.1644～0.8220 ?g,r =0.9996,the average recovery was 98.47% and RSD was 2.08%. CONCLUSION : This method is rapid,sensitive and accurate. It can be used for quality control of Wangshi baochi Pills.
ABSTRACT
Objective: To established a method for the quality standard of Yupingfeng Oral Liquid(Radix Astragali, Radix Saposhnikoviae and Rhizoma Atractylodis Macrocephalae). Methods:Radix Astragali, Radix Saposhnikoviae and Rhizoma Atractylodis Macrocephalae in oral liquid were identified by TLC. The content of astragaloside IV was determined by HPLC-ELSD. Results:The study showed that spots of samples on TLC can be well separated and the method had strong specificity. The average recovery was 97.75% and RSD was 1.31%. Conclusion:The method is simple, sensitive and accurate. It can be used for quality control of Yupingfeng Oral Liquid.