ABSTRACT
Aims To observe the effect of saccharide extracts of Yiqijianpi herb Codonopsis and Glycyrrhizae on polyamine-dependent activation of K+ channels sig-nal pathway during cell migration and to investigate their mechanism of promoting restoration in gastrointes-tinal mucosal injuries. Method The study was based on IEC-6 cell migration model. While in a normal polyamine level or polyamine was inhibited by DFMO, the effect of Codonopsis saccharide extracts and Glycyr-rhizae saccharide extracts on polyamine-dependent acti-vation of K+ channels signal pathway during cell mi-gration was observed. (1) K+ channel protein Kv1. 1 was determined by Western blot. (2)Membrane poten-tial was measured by Flow Cytometer. (3) Laser scan-ning confocal microscope was used for measuring [ Ca2+] cyt. ( 4 ) The expression of RhoA, which is Ca2+ downstream protein, was determined by Western blot. Results During cell migration, Codonopsis and Glycyrrhizae saccharide extracts could: ( 1 ) improve the expression of Kv1 . 1 protein and ameliorate the de-crease of kv1. 1 protein expression by DFMO;(2) in-crease membrane hyperpolarization and reverse mem-brane depolarization resulted by DFMO; ( 3 ) improve intracellular [ Ca2+] cyt, while Codonopsis could re-verse the decrease of [ Ca2+] cyt caused by DFMO;(4) improve the expression of RhoA protein, reversing its decline caused by DFMO. Conclusion Codonop-sis and Glycyrrhizae saccharide extracts can promote cell migration in IEC-6 cell, which is correlated with their effect on polyamine-dependent activation of K+channels signal pathway.
ABSTRACT
Objective To prepare long-circulating temperature-sensitive liposomes with paclitaxel( LTSLP ),develop methods for determination of paclitaxel,related substances and monostearoyl phosphatidylcholine( MSPC ) in LTSLP,and the haemolysis of LTSLP in vitro. Methods HPLC-UV methods for paclitaxel content and related substances and HPLC-CAD method for MSPC in LTSLP were established and validated. Spectrophotometric method was used to determine hemolysis in vitro. Results There was a good linear relationship between peak area and concentration within the range of 60. 39-181. 17μg·mL-1 . Recovery and precision of the method for determination of paclitaxel content met the requirements. Specificity,sensitivity,and system suitability for related substances were consistent with requirements. There was a good linear relationship between peak area and concentration within the range of 1. 5-50. 0μg·mL-1 for the determination of MSPC with good specificity,sensitivity and recovery. Paclitaxel contents in three batches of self-prepared LTSLP were between 90. 0% and 110. 0%,single related substances were below 0. 5% and total impurities were below 2. 0%. There was almost no hemolysis in vitro. Conclusion The methods for determining paclitaxel content,related substances and haemolysis can be used to assess the quality of LTSLP. Self-produced LTSLP consistently meet the quality standards.