Mesenchymal stem cell calcification induced by protoscolex of two species of Echinococcus: a differential analysis

doi:10.3969/j.issn.2095-4344.2119

Abstract

Abstract: Abstract
BACKGROUND: The growth pattern of Echinococcus granulosus is different from that of Echinococcus alveolaris. Hepatic echinococcosis can form a complete fibrous calcified cyst wall, while hepatic alveolar echinococcosis can grow infiltratively and cannot form a complete fibrous calcified cyst wall. Bone marrow mesenchymal stem cells (BMSCs) are involved in the formation of calcified wall of hydatidosis, but the calcification characteristics of Echinococcus granulosus and Echinococcus alveolaris are different and the role of BMSCs is still unclear.
OBJECTIVE: To compare the effects of Echinococcus granulosus and Echinococcus alveolaris on the calcification of BMSCs and to preliminarily investigate the formation mechanism of echinococcosis calcifications.
METHODS: BMSCs of C57BL/6 mice were extracted, cultured and identified, followed by co-culture with the protoscolex of Echinococcus granulosus (BMSC+CE group) and Echinococcus alveolaris (BMSC+AE group), respectively. BMSCs cultured alone were used as control group. After 1, 4, and 7 days of co-culture, alkaline phosphatase activity was detected by a microplate reader, the expression of BMP2 and RUNX2 mRNA was detected by RT-q PCR, and the expression of BMP2, RUNX2 and phosphorylated Smad1/5/8 (P-Smad1/5/8) proteins was detected by western blot assay.
RESULTS AND CONCLUSION: (1) The alkaline phosphatase activity of the BMSC+CE group and BMSC+AE group was significantly higher than that of the control group at 1 and 4 days after culture (P < 0.05), and the alkaline phosphatase activity of the BMSC+CE group was significantly higher than that of BMSC+AE group (P < 0.05). (2) Western blot results showed that the expression of BMP2, RUNX2, and P-Smad1/5/8 protein in the BMSC+CE group and BMSC+AE group was significantly higher than that in the control group at 1 and 4 days after culture (P < 0.05), while the expression of BMP2, RUNX2, and P-Smad1/5/8 protein in the BMSC+CE group was significantly higher than that in the BMSC+AE group (P < 0.05). (3) RT-qPCR results showed that the expression of BMP2 and RUNX2 mRNA in the BMSC+CE group was significantly higher than that in the control group at 1, 4 and 7 days after culture (P < 0.05), and was significantly higher than that in the BMSC+AE group at 4 and 7 days after culture (P < 0.05). The expression of RUNX2 mRNA in the BMSC+AE group was significantly higher than that in the control group at 1, 4, and 7 days after culture (P < 0.05). (4) To conclude, co-culture of the protoscolex of Echinococcus alveolaris and BMSCs promotes the expression of alkaline phosphatase and RUNX2 in BMSCs by up-regulating BMP-Smad1/5/8 pathway. At the later stage of co-culture, the effect of Echinococcus alveolaris on BMSCs calcification is significantly weakened, while the effect of Echinococcus granulosus on BMSCs calcification remains unchanged, suggesting that this mechanism may be related to the different growth patterns of two kinds of hydatids.

Key words: bone marrow mesenchymal stem cells, Echinococcus alveolaris, Echinococcus granulosus, calcification, fibrous calcified wall, protoscolex of Echinococcus

CLC Number:

Gui Xianwei, Jiang Huijiao, Wu Jie, Liang Xueqi, Xu Xiaodan, Wang Erqiang, Zou Hailiang, Chen Hejie, Chen Xueling, Wu Xiangwei. Mesenchymal stem cell calcification induced by protoscolex of two species of Echinococcus: a differential analysis[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(1): 38-43.

Figures/Tables 4

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