Experimental Study On Anti-hematologic Malignancies Of Extracts From Marsdenia Tenacissima

【Objective】To investigate the effect of anti-hematological malignances and its mechanisms of extracts from Marsdenia tenacissima(trade name:Xiao aiping),and the effect of extracts from Marsdenia tenacissima on normal human immunocytes and hemopoietic stem cells(HSCs).【Methods】MTT method was adopted to observe the inhibiting effect of extracts from Marsdenia tenacissima with 0μL/mL～100μL/mL on Jurkat,Raji,RPMI8226,HL-60,K562 cell lines for 1～5 days.Cell cycle was detected by flow cytometry(FCM).Cell morphology and apoptosis were measured by Annexin V/PI staining and mitochondrial membrane potential(ΔΨm) was measured by JC-1 assay.Western blot method was used to detect the protein expression of caspase-3.Cell proliferation was measured by MTT assay in normal cells and cells treated with ConA or LPS.Phagocytosis of normal macrophage was measured by the dose of Neutral Red Dye.CFU-GM was assessed by semi-solid culture assay for determination of colony formation capacity of HSCs.【Results】1.Extracts from Marsdenia tenacissima had the most significant inhibition on Jurkat cells with dose-dependent among Jurkat,Raji,RPMI8226,HL-60,K562 cell lines in vitro. Extracts from Marsdenia tenacissima could induce G₂ phase arrest of RPMI8226 and Raji cell lines.2.Extracts from Marsdenia tenacissima had varying degrees of apoptosis induction on Jurkat,Raji,RPMI8226 cell lines in vitro.The apoptosis rate of Jurkat cells was the highest with dose-,time- dependent.Extracts from Marsdenia tenacissima with 25 and 50μL/mL could decrease cellΔΨm and induce apoptosis in Jurkat and Raji cells,while extracts from Marsdenia tenacissima with 100μL/mL could also decrease cellΔΨm and induce apoptosis in RPMI8226 cells.No significant changes in protein expressions of caspase-3 in groups treated by extracts from Marsdenia tenacissima.3.Extracts from Marsdenia tenacissima could enhance proliferation of normal lymphocytes treated with ConA or LPS with dose-dependent in vitro.There was no effect of extracts from Marsdenia tenacissima on macrophage to ingest Neutral Red Dye and CFU-GM colony formation capacity of normal bone marrow/ peripheral HSCs.【Conclusion】1.Extracts from Marsdenia tenacissima have varying degrees of inhibition on various hematological malignance cell lines in vitro.2.Extracts from Marsdenia tenacissima induce apoptosis on some lymphocytic leukemic cell strains and myeloma cell strain in vitro by mitochondrial pathway.3.There is no significant cell toxicity of extracts from Marsdenia tenacissima on normal immunocytes and HSCs in vitro.And extracts from Marsdenia tenacissima can enhance proliferation of normal lymphocytes treated with ConA or LPS in vitro.