ABSTRACT
OBJECTIVE：To comp are cytotoxicity and anti-inflammatory effects of raw Aconitium kusnezoffii and A. kusnezoffii processed with Terminalia chebula . METHODS ：Using H 9c2 cardiomyocytes isolated from rat as subjects ，CCK-8 assay was used to detect the effects of 0.5，1，2，4，6，8，10 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula on cell inhibition rate after cultured for 4，8，12，24 h. Hoechst 33258 staining was used to observe the effects on cell morphology characteristics after treated with 2，4，6 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula . Using macrophages RAW264.7 cells as subjects ，CCK-8 assay was used to detect the effects of 0.05，0.1，0.25，0.5，0.75，1，1.5，2 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula on cell survival rate after cultured for 24 h. ELISA assay was used to detect the effects of 0.05，0.1，0.25，0.5 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula on the release of NO ， TNF-α and IL-6 in RAW 264.7 inflammation cells induced by LPS. RESULTS ：When the mass concentration was 0.5，1 mg/mL， neither raw A. kusnezoffii and A. kusnezoffii processed with T. chebula had no inhibitory effect on H 9c2 cells. When the mass concentration was 2 mg/mL，the inhibitory effects of A. kusnezoffii processed with T. chebula on H 9c2 cells was higher than that of raw A. kusnezoffii （P＜0.05 or P＜0.01）；fluorescence intensity of cells treated for 24 h was stronger than that of raw A. kusnezoffii，but its nucleus was intact. When the mass concentration was 4-10 mg/mL，the inhibitory rate of A. kusnezoffii processed with T. chebula on H 9c2 cells at different time points （except for 24 h culture of 8，10 mg/mL）was significantly lower than raw A. kusnezoffii （P＜0.05 or P＜0.01）. The characteristics of cell morphology also showed that the fluorescence intensity of raw A. kusnezoffii group at 4，6 mg/mL was stronger than that of A. kusnezoffii processed with T. chebula group，and the cell nucleus fragmentation was more serious in the raw A. kusnezoffii group. 0.05-0.5 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula had no toxicity to RAW264.7 cells. 0.25，0.5 mg/mL raw A. kusnezoffii and 0.1，0.25，0.5 mg/mL A. kusnezoffii processed with T. chebula showed significant inhibitory effect on the release of NO ，0.05，0.1，0.25，0.5 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula showed significant inhibitory effect on the release of TNF-α and IL-6 in RAW 264.7 cell（P＜0.05 or P＜ 0.01）. The inhibitory effects of A. kusnezoffii processed with T. chebula at the same concentration on the release of NO was better than that of raw A. kusnezoffii ，but poorer than raw A. kusnezoffii in the inhibitory effects on the release of TNF-α and IL-6. CONCLUSIONS：The toxicity of A. kusnezoffii can be reduced after processed with T. chebula ，especially the toxicity of it in medium or high concentration and short-term use is lower than that of raw A. kusnezoffii . At the same time ，the anti-inflammatory effect of A. kusnezoffii processed with T. chebula is comparable to that of raw A. kusnezoffii at the same concentration.