Objective: To investigate the effects of medicated serum of Hei Dihuangwan (MSHDW) on autophagy and paracrine secretion in bone marrow stromal cells (BMSCs) and explore the mechanism of Hei Dihuangwan (HDW) on renal anemia. Method: Rat models of renal anemia were established

and then normal and renal anemia BMSCs were separated by using gradient density centrifugation and adherent culture method. The third generation BMSCs were used and after starvation induction for 1 h

they were divided into normal group

model group

HDW high dose

middle dose and low dose groups. Cell survival rate was detected by MTT method; electron microscopy was used to observe the autophagosomes; the mRNA expression levels of LC3Ⅰ

LC3Ⅱ

and Becline-1 were determined by Real-time PCR. ELISA was used to detect stem cell factor (SCF)

erythropoietin (EPO)

and insulin-like growth factor (IGF)-1 in culture supernatant. Finally

the correlation between autophagy genes and hematopoietic factors were investigated by using canonical correlation analysis. Result: As compared with normal group

BMSCs survival rate was significantly decreased in model group (P<0.01); the cell survival rates in HDW high dose

middle dose and low dose groups were higher than that in model group (P < 0.05

P < 0.01)

and the effect was enhanced with the increase of dose. Under the electron microscope

the number of autophagosomes in model group was larger than that in normal group; and the number in HDW various groups was all smaller than that in model group; mRNA expression level of autophagy related genes in model group was higher than that in the normal group (P<0.01); mRNA expression levels of autophagy related genes in HDW high dose and middle dose groups were lower than those in model group (P < 0.05

P < 0.01). Hematopoietic factors in culture supernatant of model group BMSCs were less than those in normal group (P<0.01); the levels in HDW high dose and middle dose groups were higher than those in model group (P < 0.05

P < 0.01)

and the effect was enhanced with the increase of dose. Canonical correlation analysis showed that

autophagy genes of BMSCs were highly positively correlated to inhibition of paracrine secretion under MSHDW effect. Conclusion: The mechanism of HDW in treating renal anemia may be related to down-regulating the levels of autophagy in BMSCs and promoting paracrine secretion hematopoietic factors.