Effects of bone marrow mesenchymal cells immune thrombocytopenia patients on the biological behaviors of megakaryocytes
2018, 38 (6):
Objective · To investigate changes of immune thrombocytopenia (ITP) patients-derived bone marrow mesenchymal cells (BMCs) in cells survival, cytokines as well as the effects of BMCs on the biological behaviors of megakaryocytes. Methods · BMCs were collected 7ITP patients and 5 normal controls (NC), and cultivatedthe whole marrow adherent method. Surface markers and basal apoptosis rate of BMCs were analyzedflow cytometry (FCM). Proliferation of BMCs was assessedCCK-8 method. Phorbol 12-myristate 13-acetate (PMA) was used to stimulate differentiation of HEL cells. The induced HEL cells (inHEL) were divided into 3 groups: inHEL cultured alone (group a), inHEL co-cultured with BMCs derived ITP patients (group b), inHEL co-cultured with BMCs derived NC (group c). After 72 h incubation, the of cell surface proteins (CD41a, CD42b) and cell apoptosis rate were analyzedFCM. The mRNA and proteins levels of cytokines IL6, IL11, TPO, SCF were detectedreal-time fluorescent quantitative PCR (RT-qPCR) and enzyme linked immunosorbent assay (ELISA), respectively. Results · Compared with NC, BMCs ITP patients grew progressively slowly (Day 4, P0.039; Day 6, 10, P0.009; Day 8, P0.007), cell basal apoptosis rates were increased [AV+PI-(early apoptosis rate), P0.036; AV+PI+ (late apoptosis rate), P0.003; AV+PI-/+ (total apoptosis rate), P0.004]. Compared with group a, the of CD41a in group c was much higher (P0.000). The of CD41a in group b was higher than that in group a (P0.015), but still much less than that in group c (P0.000). Compared with group a, the early and total apoptosis rate in group b, c and the late apoptosis rate in groupc were decreased obviously (all P0.000), whereas there was no obvious change of the late apoptosis rate in group b. However, compared with group c, the late and total apoptosis rate in group b were significantly increased (both P0.000). The levels of IL6, SCF mRNA and IL6 protein were significantly decreased in ITP BMCs (all P0.000), but there was no obvious difference in the levels of IL11 and TPO between ITP BMCs and NC BMCs. Conclusion · BMCs ITP patients show some defects in supporting megakaryocytic differentiation and survival under co-culture conditions, which mechanisms are related to the reduction of IL6 and SCF .
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