Protective Effect and Mechanism of Matrine Combined with Glycyrrhizic Acid in the Treatment of Chronic Liver Injury Induced by Carbon Tetrachloride / 中国药师

Objective: To investigate the protective effects of matrine combined with glycyrrhizic acid on chronic liver injury induced by carbon tetrachloride, and explore the protective mechanism from the points of energy metabolism and CYP enzyme.Methods: The chronic hepatic injury model of rats was induced by CCl4.The changes of activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were measured to observe the protective effect of the two drugs and their combination.The contents of glutamate dehydrogenase (GLDH) in serum and adenine nucleoside three phosphate (ATP), adenosine diphosphate (ADP) and adenine monophosphate (AMP) in liver tissue were determined to evaluate the regulation effect on hepatic energy metabolism and mitochondrial function.The levels of CYP1A2, CYP2E1 mRNA and protein in liver tissue were detected by real-time PCR and Western Blot to evaluate the two drugs and their combination on the regulation function of liver CYP enzyme.Results: Matrine (72.8 mg×kg-1)and glycyrrhizic acid(43.4 mg·kg-1)could decrease the serum activities of ALT and ALT in chronic hepatic injury model, and the combination (matrine 36.4 mg·kg-1+glycyrrhizic acid 21.7 mg·kg-1) had the most significant protective effect (P＜0.05).Matrine (72.8 mg·kg-1)and glycyrrhizic acid(43.4 mg·kg-1)could decrease GLDH in serum,and restore the content of ATP in liver (P＜0.05).Matrine (72.8 mg·kg-1) had no effect on the expression of CYP1A2 and CYP2E1mRNA, and glycyrrhizin (43.4 mg·kg-1) could inhibit the expression of CYP1A2, CYP2E1mRNA and protein (P＜0.05).Conclusion: Matrine combined with glycyrrhizin has obvious regulation effect on mitochondrial function and liver protective effect in chronic hepatic injury model.

Inhibitory effect of cyclosporin A on growth and collagen synthesis of rat cardiac fibroblasts induced by arginine vasopressin / 药学学报

Aim To investigate the effects of cyclosporin A (CsA) on growth and collagen synthesis of cardiac fibroblasts (CFs) induced by arginine vasopressin (AVP). Methods CFs of neonatal Sprague-Dawley rats were isolated by trypsinization and cultured; growth-arrested CFs were stimulated with 1×10-7 mol·L-1 AVP in the presence or absence of CsA (0.05, 0.5 and 5 μmol·L-1). MTT and flow cytometry techniques were adopted to measure cell number and analyze cell cycle respectively. Collagen synthesis was determined by measurement of hydroxyproline content in culture supernatant with colorimetry. Calcineurin activity was estimated by chemiluminescence. Trypan blue staining to test the viability of CFs. Results 0.05, 0.5 and 5 μmol·L-1 CsA inhibited the increase of CFs number induced by 1×10-7 mol·L-1 AVP in a dose-dependent manner, with the inhibitory rates by 12%, 24% and 29%, respectively (P＜0.05). Furthermore, cell cycle analysis showed 0.5 μmol·L-1 CsA decreased the S stage percentage and proliferation index of CFs stimulated by AVP (P＜0.05). In culture medium, the hydroxyproline content induced by AVP decreased by 0.5 and 5 μmol·L-1 CsA (P＜0.05), with the inhibitory rates of 29% and 33%, respectively. CsA completely inhibited the increment of calcineurin activity induced by AVP (P＜0.01), but CsA itself had no effect on the baseline of calcineurin activity and CFs viability. Conclusion CsA inhibits proliferation and collagen synthesis of CFs by virtue of blocking calcineurin signaling pathway and might provide a novel target for prevention and treatment to cardiac fibrosis.

Induction of apoptosis by Tryptanthrin on K562 cells / 中国药理学通报

Aim To study the effect of Tryptanthrin(Try) on proliferation and apoptosis of erythroleukemia K562 cells.Methods The cell proliferation effect of Try(1.56～50 mg?L-1) on K562 cells was assessed by MTT assay.The morphologic change was observed by Hoechst 33258 fluore-scent stain.The flow cytometer was used to detect cell apoptosis and cell cycle.Results MTT showed that in the range of 3.12～50 mg?L-1 Try obviously inhibited the proliferation of K562 cells in a dose and time-dependent manner.Typical apoptosis changes were observed in K562 cells treated with Try for 48 h by flourescence inverted microscope.With Annexin V-FITC and PI double staining,folw cytometer result showed that the apoptosis state was obvious in K562 cells treated with 25,50 mg?L-1 Try for 48 h.The cell cycle distribution of K562 was changed.The G0/G1 phase was blocked and the DNA synthesis was inhibited,accompanied with subdiploid apoptotic peak.Conclusion Try has an effect on inhibiting the cell proliferation and inducing the apoptosis of K562.