The effect of koumine on cartilage injury in rats with knee osteoarthritis by regulating AMPK/NLRP3 pathway

doi:10.11958/20252961

Abstract

Abstract:

Objective To discuss the effect of koumine (KM) on cartilage injury in rats with knee osteoarthritis (KOA) by regulating the adenosine monophosphate activated protein kinase (AMPK)/Nod-like receptor protein 3 (NLRP3) pathway. Methods The KOA rat model was constructed and separated into the KOA group, the low and high dose KM (L, H-KM) groups, the positive drug (celecoxib) group and the H-KM+AMPK inhibitor Compound C group, with 12 rats per group. Another 12 rats were used as the control (CK) group. The knee joint width, joint swelling and gait score were measured. ELISA was used to detect the levels of IL-18, IL-1β and TNF-α. HE staining, Safranin O-Fast Green staining and transmission electron microscopy were used to observe the pathological changes, structural changes and microstructure of cartilage tissue. In addition, Western blot assay was used to detect the expression of AMPK/NLRP3 pathway related proteins. Results The KOA group showed obvious pathological changes: rough tissue surface, disordered cell hypertrophy and broken tidal lines. The lightening of the red color in Safran-solid green staining indicated the loss of proteoglycans. The cells experienced particle shedding, swelling and rupture of their contents. The KOA group showed increased knee joint width, joint swelling degree, gait score, IL-18, IL-1β, TNF-α levels and NLRP3 protein expression, while decreased p-AMPK/AMPK ratio than those of the CK group (P<0.05). The L-KM group and the H-KM group and the celecoxib group showed decreased knee joint width, joint swelling degree, gait score, IL-18, IL-1β, TNF-α levels, NLRP3 protein expression and pathological damage, while increased p-AMPK/AMPK ratio than those of the KOA group (P<0.05). The H-KM+Compound C group showed increased knee joint width, joint swelling degree, gait score, IL-18, IL-1β, TNF-α levels, NLRP3 protein expression and pathological damage, while decreased p-AMPK/AMPK ratio than those of the CK group (P<0.05). Conclusion KM may alleviate cartilage injury in KOA rats by activating the AMPK/NLRP3 pathway.

Key words: gelsemine, osteoarthritis, knee, AMP-activated protein kinases, NLR family, pyrin domain-containing 3 protein, cartilage

CLC Number:

R684.3

Figures/Tables 7

References 21

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组别	膝关节宽度/mm	关节肿胀度/%	步态评分/分
CK组	4.69±0.56	0.00±0.00	0.00±0.00
KOA组	9.15±0.93^a	36.52±3.71^a	2.10±0.22^a
L-KM组	7.83±0.81^b	23.67±2.89^b	1.30±0.18^b
H-KM组	5.27±0.61^bc	11.05±1.53^bc	0.50±0.13^bc
塞来昔布组	5.24±0.59^bc	11.86±1.58^bc	0.40±0.09^bc
H-KM+Compound C组	8.22±0.85^d	28.37±3.02^d	1.70±0.20^d
F	78.210^＊＊	355.539^＊＊	335.802^＊＊

组别	膝关节宽度/mm	关节肿胀度/%	步态评分/分
CK组	4.69±0.56	0.00±0.00	0.00±0.00
KOA组	9.15±0.93^a	36.52±3.71^a	2.10±0.22^a
L-KM组	7.83±0.81^b	23.67±2.89^b	1.30±0.18^b
H-KM组	5.27±0.61^bc	11.05±1.53^bc	0.50±0.13^bc
塞来昔布组	5.24±0.59^bc	11.86±1.58^bc	0.40±0.09^bc
H-KM+Compound C组	8.22±0.85^d	28.37±3.02^d	1.70±0.20^d
F	78.210^＊＊	355.539^＊＊	335.802^＊＊

组别	IL-18	IL-1β	TNF-α
CK组	96.58±12.55	49.21±6.15	102.54±13.62
KOA组	143.78±14.62^a	78.94±7.95^a	150.73±15.53^a
L-KM组	124.69±13.89^b	64.85±7.11^b	123.56±14.26^b
H-KM组	104.25±13.21^bc	53.18±6.83^bc	107.58±14.05^bc
塞来昔布组	103.16±13.10^bc	52.37±6.79^bc	105.16±13.87^bc
H-KM+Compound C组	133.91±14.25^d	71.36±7.63^d	139.18±15.04^d
F	23.671^＊＊	34.083^＊＊	23.015^＊＊

组别	IL-18	IL-1β	TNF-α
CK组	96.58±12.55	49.21±6.15	102.54±13.62
KOA组	143.78±14.62^a	78.94±7.95^a	150.73±15.53^a
L-KM组	124.69±13.89^b	64.85±7.11^b	123.56±14.26^b
H-KM组	104.25±13.21^bc	53.18±6.83^bc	107.58±14.05^bc
塞来昔布组	103.16±13.10^bc	52.37±6.79^bc	105.16±13.87^bc
H-KM+Compound C组	133.91±14.25^d	71.36±7.63^d	139.18±15.04^d
F	23.671^＊＊	34.083^＊＊	23.015^＊＊

组别	p-AMPK/AMPK	NLRP3/β-actin
CK组	0.87±0.09	0.46±0.05
KOA组	0.49±0.05^a	0.82±0.09^a
L-KM组	0.61±0.06^b	0.71±0.08^b
H-KM组	0.79±0.08^bc	0.63±0.07^bc
塞来昔布组	0.82±0.08^bc	0.51±0.06^bc
H-KM+Compound C组	0.53±0.06^d	0.76±0.08^d
F	30.859^＊＊	22.725^＊＊