Xiao Chai Hu Tang In Vitro Induction Of Hl-60 Cell Differentiation And Its Mechanism

Objective: To observe the effects of HL-60 cell differentiation induced by Xiaocaihu decoction and to discuss its mechanism preliminarily.Methods: To cultivate respectively the HL-60 cells in vitro with the xiaocaihu decoction of different dosage for 6 days, the cells differentiation was observed by some changes of cytomorphology, the cell swallow function, NBT reducing ability, cell surface differentiation antigen CD11b, CD14 expression and etc. According to the changes of cell cycle by flow cytometry and anti-oncogene P⁵³ expression by immunity histochemistry, the mechanism was revealed preliminarily. And to compare xiaocaihu's function to ATRA which was supposed as positive control group.Result:1. The changes of cytomorphology: By middle-dose and high-dose Xiaochaihu decoction role, the part of the cells differentiation were induced into mature granulocyte morphologically; Compared to the blank control group, in middle-dose and high-dose Xiaochaihu decoction groups, it was observed that the number of myelocytes and metamyelocytes increased, specially myelocytes amount, the number of band cells and segmented cells changed insignificantly, and promyelocytes decreased; In addition, the proportion of their myelocytes, metamyelocytes, band cells and segmented cells was lower than those in ATRA group.2. The changes of cell function and biochemical traits: Compared to the blank control group, HL-60 cell phagocytosis and NBT reduction ability had a tendency to be strong in low-does Xiaocaihu decoction group, but without statistical significance (P> 0.05); And in middle-dose or high-dose Xiaochaihu decoction, the both above were enhanced markedly. There was significant difference (P <0.05). However, enhancing role of two different dose Xiaochaihu decoction above was weaker than ATRA, with the significant difference (P <0.05).3. The changes of CD11b, CD14 expression: Compared to the blank control group, CD11b and CD14 expression were no statistical significance by different does Xiaocaihu decoction (P> 0.05). And they were enchanced markedly by ATRA.4. The changes of cell cycle: Compared to the blank control group, HL—60 cell cycle was blocked into G₀/G₁ phase and the cells amount in S phase decreased in middle-dose and high-dose Xiaochaihu decoction groups.5. the changes of P⁵³ expression: compared to the blank control group, P⁵³ expression changed insignificantly (P>0.05) in low-does and middle-does xiaocaihu decoction groups; on the contrary, P⁵³ expression was enhanced significantly (P<0.05) in high-does xiaocaihu decoction group. The enhancing P⁵³ expression role of different dose Xiaochaihu decoction was weaker than ATRA, with the significant difference (P <0.05).Conclusion: HL—60 cells can be differentiation induced into mature granulocyte by xiaocaihu decoction, but their differentiation effects are poorer than ATRA; This mechanism may be a result of blocking cells into G₀/G₁ phase and increasing P⁵³ expression; In addition, the CD11b expression does not increase correspondingly.