Effects of High active antiretroviral therapy on the osteogenesis of bone marrow mesenchymal stem cell

doi:10.3877/cma.j.issn.1674-1358.2017.04.002

Abstract

Abstract:

Objective

To investigate the intervention of three high active antiretroviral therapy (HAART) drugs serum on proliferation and osteogenic differentiation of bone mesenchymal stem cell (BMSCs), and explore the effects of HAART drugs on bone metabolism.

Methods

Rats’ serums containing three kinds of HAART drug were prepared to intervene the culture and differentiation of BMSCs. Proliferation of BMSCs was measured by CCK assay. Alkaline phosphatase, calcium alizarin and oil red O staining were used to observed the differentiation of cells osteogenesis and adipogenesis. The levels of gene expression of Runx 2 and osteoprotegerin (OPG) in the osteogenetic differentiation of BMSCs were tested by RT-PCR.

Results

Compared with the rat serum group, A_{72 h} value of three kinds of drug serum group were significantly different (F = 15.42, P = 0.004). Compared with the control group, alkaline phosphatase positive cells which reflected osteogenesis differentiation decreased in drug serum group, and calcium nodules decreased in alizarin red staining test, in which tenofovir disoproxil containing group was the most significantly secreased. Fat cells which reflect adipogenic induction increased. In PCR detection, compared with control group, the expression of Runx 2 decreased and OPG expression increased in tenofovir disproxil containing group (F = 11.81, P = 0.026; F = 10.14, P = 0.033).

Conclusions

HAART drugs suppressed the proliferation and osteogenic differentiation of BMSCs which the influence of tenofovir disoproxil was most significant.

Key words: High active antiretroviral therapy, Bone mesenchymal stem cell, Osteoblast

Xuemin Quan, Qiang Zhang, Zhengrong Gao, Changsong Zhao, Rugang Zhao, Jingjing Wang, Kun Liu, Zongfeng Chen. Effects of High active antiretroviral therapy on the osteogenesis of bone marrow mesenchymal stem cell[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2017, 11(04): 320-325.

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References 25

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基因		序列（3'→5'）	产物大小（bp）
GAPDH			228
	F	GGCAAGTTCAACGGCACAG
	R	CGCCAGTAGACTCCACGACA
OPG			152
	F	CGGATGGGTTCTTCTCAGGT
	R	GCACAGGGTGACATCTATTCCA
Runx 2			130
	F	TGCTTCATTCGCCTCACAAAC
	R	TTGCAGCCTTAAATGACTCGG

基因		序列（3'→5'）	产物大小（bp）
GAPDH			228
	F	GGCAAGTTCAACGGCACAG
	R	CGCCAGTAGACTCCACGACA
OPG			152
	F	CGGATGGGTTCTTCTCAGGT
	R	GCACAGGGTGACATCTATTCCA
Runx 2			130
	F	TGCTTCATTCGCCTCACAAAC
	R	TTGCAGCCTTAAATGACTCGG

分组	0 h	24 h	48 h	72 h
胎牛血清	0.169 ± 0.026	0.298 ± 0.057	0.492 ± 0.051	0.787 ± 0.066
大鼠血清	0.170 ± 0.024	0.273 ± 0.030	0.373 ± 0.047	0.516 ± 0.020
3TC药物血清	0.173 ± 0.029	0.289 ± 0.040	0.369 ± 0.022	0.527 ± 0.031
TDF药物血清	0.171 ± 0.024	0.277 ± 0.034	0.343 ± 0.022	0.455 ± 0.022^a
LPV/r药物血清	0.169 ± 0.024	0.285 ± 0.041	0.379 ± 0.018	0.488 ± 0.018
DMEM	0.176 ± 0.026	0.202 ± 0.022	0.225 ± 0.062	0.235 ± 0.029

分组	0 h	24 h	48 h	72 h
胎牛血清	0.169 ± 0.026	0.298 ± 0.057	0.492 ± 0.051	0.787 ± 0.066
大鼠血清	0.170 ± 0.024	0.273 ± 0.030	0.373 ± 0.047	0.516 ± 0.020
3TC药物血清	0.173 ± 0.029	0.289 ± 0.040	0.369 ± 0.022	0.527 ± 0.031
TDF药物血清	0.171 ± 0.024	0.277 ± 0.034	0.343 ± 0.022	0.455 ± 0.022^a
LPV/r药物血清	0.169 ± 0.024	0.285 ± 0.041	0.379 ± 0.018	0.488 ± 0.018
DMEM	0.176 ± 0.026	0.202 ± 0.022	0.225 ± 0.062	0.235 ± 0.029

组别	Runx 2 mRNA	OPG mRNA
大鼠血清	1.000 ± 0.170	1.000 ± 0.194
3TC药物血清	0.962 ± 0.152	1.085 ± 0.056
TDF药物血清	0.620 ± 0.089	1.414 ± 0.114
LPV/r药物血清	0.920 ± 0.089	1.131 ± 0.055

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