Impacts of alpinetin on angiogenesis in knee osteoarthritis rats by regulating the VEGF/SphK1/S1P signaling pathway

doi:10.11958/20230892

Abstract

Abstract:

Objective To investigate effects of alpinetin (APT) on angiogenesis in knee osteoarthritis (KOA) rats by regulating vascular endothelial growth factor/sphingosine kinase 1/sphingosine 1 phosphate (VEGF/SphK1/S1P) signaling pathway. Methods The KOA rat model was established by Videman method. Ninety rats were grouped into the control group, the model group, the low-dose kaempferol group (L-APT group), the high-dose kaempferol group (H-APT group), the high-dose kaempferol group+lentivirus negative control group (APT+NC group) and high-dose kaempferol+overexpression of SphK1 lentivirus group (APT+SphK1 group), with 15 rats in each group. Pathological changes of cartilage tissue in rats were observed by HE staining. Contents of IL-1β, TNF-α, IL-6 and MMP-13 in cartilage tissue were measured by enzyme linked immunosorbent assay. Chondrocyte apoptosis of cartilage tissue cells was detected by TUNEL. VEGF and CD31 protein positive expression levels were detected by immunohistochemistry assay. The p-VEGFR2, VEGFR2, SphK1 and S1P protein levels were detected by Western blot assay. Results Rats in the model group showed pathological damage. Compared with the control group, the apoptosis rate, IL-1β, TNF-α, IL-6, MMP-13 levels, VEGF positive expression, CD31 positive expression, p-VEGFR2, SphK1 and S1P protein expression levels were increased in the model group (P<0.05). Compared with the model group, the pathological damage was obviously reduced in the L-APT group and the M-APT group, and cell apoptosis rate, IL-1β, TNF-α, IL-6, MMP-13 levels, VEGF positive expression, CD31 positive expression, p-VEGFR2, SphK1 and S1P protein expression levels were obviously reduced (P<0.05). Compared with the APT+NC group, the pathological injury of cartilage tissue increased in the APT+SphK1 group, cell apoptosis rate, IL-1β, TNF-α, IL-6, MMP-13 levels, VEGF positive expression, CD31 positive expression, p-VEGFR2, SphK1 and S1P protein expression levels were obviously increased (P<0.05). Conclusion APT inhibits angiogenesis in knee osteoarthritis rats by inhibiting the VEGF/SphK1/S1P signaling pathway.

Key words: osteoarthritis, knee, neovascularization, pathologic, vascular endothelial growth factors, alpinetin, sphingosine kinase 1, sphingosine 1 phosphate

CLC Number:

R684.3

Figures/Tables 7

References 24

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组别	IL-1β/（ng/L）			TNF-α/（ng/L）
Control组	38.41±2.73			93.24±5.37
Model组	147.36±10.34^a			327.59±10.34^a
L-APT组	121.29±9.57^b			243.37±9.52^b
H-APT组	59.44±4.25^bc			125.46±6.25^bc
APT+NC组	55.38±4.19			120.52±6.13
APT+SphK1组	114.61±6.83^d			223.66±8.24^d
F	201.238^＊＊			667.445^＊＊
组别		IL-6/（ng/L）	MMP-13/（μg/L）
Control组		5.32±0.94	29.38±1.63
Model组		34.27±5.34^a	246.46±24.16^a
L-APT组		28.49±3.17^b	164.62±17.39^b
H-APT组		13.51±2.05^bc	105.29±12.25^bc
APT+NC组		12.26±1.65	98.57±11.34
APT+SphK1组		22.18±2.26^d	148.33±15.27^d
F		69.389^＊＊	114.780^＊＊

组别	IL-1β/（ng/L）			TNF-α/（ng/L）
Control组	38.41±2.73			93.24±5.37
Model组	147.36±10.34^a			327.59±10.34^a
L-APT组	121.29±9.57^b			243.37±9.52^b
H-APT组	59.44±4.25^bc			125.46±6.25^bc
APT+NC组	55.38±4.19			120.52±6.13
APT+SphK1组	114.61±6.83^d			223.66±8.24^d
F	201.238^＊＊			667.445^＊＊
组别		IL-6/（ng/L）	MMP-13/（μg/L）
Control组		5.32±0.94	29.38±1.63
Model组		34.27±5.34^a	246.46±24.16^a
L-APT组		28.49±3.17^b	164.62±17.39^b
H-APT组		13.51±2.05^bc	105.29±12.25^bc
APT+NC组		12.26±1.65	98.57±11.34
APT+SphK1组		22.18±2.26^d	148.33±15.27^d
F		69.389^＊＊	114.780^＊＊

组别	VEGF	CD31
Control组	1.78±0.24	3.21±0.24
Model组	30.36±2.47^a	14.36±0.96^a
L-APT组	21.19±2.03^b	11.15±0.73^b
H-APT组	7.41±0.84^bc	6.58±0.61^bc
APT+NC组	7.24±0.79	6.33±0.55
APT+SphK1组	18.27±1.36^d	9.69±0.68^d
F	256.424^＊＊	179.247^＊＊

组别	VEGF	CD31
Control组	1.78±0.24	3.21±0.24
Model组	30.36±2.47^a	14.36±0.96^a
L-APT组	21.19±2.03^b	11.15±0.73^b
H-APT组	7.41±0.84^bc	6.58±0.61^bc
APT+NC组	7.24±0.79	6.33±0.55
APT+SphK1组	18.27±1.36^d	9.69±0.68^d
F	256.424^＊＊	179.247^＊＊

组别	p-VEGFR2/VEGFR2	SphK1	S1P
Control组	0.52±0.03	0.44±0.02	0.33±0.03
Model组	1.00±0.06^a	1.04±0.07^a	0.93±0.08^a
L-APT组	0.89±0.04^b	0.91±0.05^b	0.79±0.05^b
H-APT组	0.71±0.05^bc	0.74±0.04^bc	0.65±0.04^bc
APT+NC组	0.69±0.04	0.71±0.05	0.62±0.03
APT+SphK1组	0.82±0.05^d	0.87±0.06^d	0.71±0.07^d
F	67.221^＊＊	83.052^＊＊	70.494^＊＊