Quality Control Of Cilian Capsule And Its Inhibitory Effect On Proliferation Of Breast Cancer Cells And Its Mechanism

Objective To establish quality standards for Cilian Capsule.Explore the prescription’s inhibitory effect on breast cancer MDA-MB-231 cell proliferation and its mechanism.Methods Radix Bupleuri,Pericarpium Citri Reticulatae Viride,Radix Paeoniae Alba and Radix Sophorae Tonkinensis were identified by thin layer chromatography(TLC),paeoniflorin and hesperidium were determined by high performance liquid chromatography(HPLC),and alcohol extract of Cilian Capsule(3.2,1.6,0.8,0.4,0.2)by MTT(mg/ml)inhibited the activity of MDA-MB-231 cells.The expression of Caspase-3 and Caspase-8 genes was detected by q RT-PCR.Results The TLC identification method of Radix Bupleuri,Pericarpium Citri Reticulatae Viride,Radix Paeoniae Alba and Radix Sophorae Tonkinensis in Cilian Capsule formula was established.The results showed that the spots of the four herbs were clear,the separation effect were good and there was no negative interference.This method has the advantages of good repeatability,simplicity,convenience,fast and reliable,low price and strong specificity,which can be used as the quality of Cilian Capsule method of qualitative identification with control.The quality control of Pericarpium Citri Reticulatae Viride and Radix Paeoniae Alba in the prescription was studied by HPLC,and the experimental results of good precision,stability and repeatability were obtained.In the range of 0.4904 μg-1.716 μg,the linear relationship of paeoniflorin peak area was good,the average recovery rate was 99.67%,RSD was 1.53%,and the linear relationship of hesperidium peak area was good in the range of 0.1183 μg-5.9125 μg,the average recovery rate was 98.06%,RSD was 1.59%.The results of in vita study showed that Cilian Capsule had a significant inhibitory effect on the growth of MDA-MB-231 cells(P<0.05)in a dose-dependent and time-dependent manner compared with the normal group,and the inhibitory effect of Cilian Capsule on MDA-MB-231 cells was the strongest with a survival rate of(38.44 ± 3.58)% 48 hours after the drug concentration was 3.2 mg/ml.The expression of Caspase-3 and Caspase-8 in MDA-MB-231 cells was(1.76±0.16)% and(1.77±0.11)%,respectively.Compared with the normal group,the expression of Caspase-3 and Caspase-8 increased with the increase of the concentration of Cilian Capsule in MDA-MB-231 cells(P<0.05).Conclusion The TLC identification characteristics of Radix Bupleuri,Pericarpium Citri Reticulatae Viride,Radix Paeoniae Alba and Radix Sophorae Tonkinensis in Cilian Capsule formula are strong.The determination method of Pericarpium Citri Reticulatae Viride and Radix Paeoniae Alba in the formula is simple and reproducible,and the identification method can control the quality of Cilian Capsule.Cilian Capsule has significant proliferation inhibition and apoptosis induction effect on MDA-MB-231 cells of breast cancer,which may be related to Capsese-3 is related to the increase in Caspase-8 expression.