The mechanism of total flavonoids of Rhizoma drynariae improving lipid metabolism disorders in steroid-induced avascular necrosis of femoral head in rabbits

doi:10.11958/20252085

Abstract

Abstract:

Objective To explore the mechanism of action of total flavonoids of Rhizoma drynariae (TFRD) in improving lipid metabolism disorders in rabbits with avascular necrosis of femoral head (SANFH) based on the peroxisome proliferator-activated receptor γ (PPARγ)/adenosine monophosphate-activated protein kinase (AMPK) pathway. Methods Sixty New Zealand rabbits were randomly divided into the control group (n=10) and the model group (n=50). The rabbit SANFH model was established by injecting Escherichia coli endotoxin and methylprednisolone. The control group was injected with the same amount of physiological saline. The SANFH model was randomly classified into five subgroups: the SANFH group, the TFRD-low group (30 mg/kg), the TFRD-medium group (60 mg/kg), the TFRD-high group (100 mg/kg) and the TFRD-high+inhibitor (TFRD 100 mg/kg + AMPK inhibiter 30 mg/kg) group, with 10 rabbits in each subgroup. After the intervention, venous blood sample was collected from the rabbit ear margin, and serum lipid levels [high-density lipoprotein cholesterol (HDL-C) and total cholesterol, triglycerides (TG)] were detected. Micro-CT was used to detect bone volume/total volume (BV/TV) (%) and trabecular quantity (Tb.N). Hematoxylin-eosin (HE) staining was used to observe histopathological changes of femur and calculate empty bone lacuna rate. TUNEL was used to detect apoptosis of femoral tissue. Real-time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR) was employed to analyze the expression of B-cell lymphoma/leukemia-2 (Bcl-2) and Bcl-2-related X protein (Bax) mRNA. Western blot assay was used to detect the expression of tissue-related proteins (PPARγ and p-AMPK/AMPK). Results Compared with the control group, HDL-C, BV/TV, Tb.N, Bcl-2 mRNAs, PPARγ and p-AMPK/AMPK were decreased in the SANFH group, and cholesterol, TG, apoptosis rate, empty bone lacuna rate and Bax mRNA were increased (P<0.05). Compared with the SANFH group, HDL-C, BV/TV, Tb.N, Bcl-2 mRNAs, PPARγ and p-AMPK/AMPK were increased in sequence in the TFRD-low group, the TFRD-medium group and the TFRD-high group, and cholesterol, TG, apoptosis rate, empty bone lacuna rate and Bax mRNA were decreased in turn (P<0.05). Compared with the TFRD-high group, changes in the above indicators were reversed in the TFRD-high+inhibitor group (P<0.05). Conclusion TFRD improves lipid metabolism disorder in SANFH rabbits by activating PPARγ/AMPK pathway.

Key words: femur head necrosis, hormones, lipid metabolism disorders, drynariae rhizoma, flavanoids, PPAR gamma, AMP-activated protein kinases

CLC Number:

R285.5

Figures/Tables 10

References 26

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基因名称	引物序列（5'→3'）	产物大小/bp
Bax	上游：CCCGAGAGGTCTTTTTCCGAG	152
Bax	下游：CCAGCCCATGATGGTTCTGAT	152
Bcl-2	上游：GGTGGGGTCATGTGTGTGG	197
Bcl-2	下游：CGGTTCAGGTACTCAGTCATCC	197
β-actin	上游：TCAGGTCATCACTATCGGCAAT	134
β-actin	下游：AAAGAAAGGGTGTAAAACGCA	134

基因名称	引物序列（5'→3'）	产物大小/bp
Bax	上游：CCCGAGAGGTCTTTTTCCGAG	152
Bax	下游：CCAGCCCATGATGGTTCTGAT	152
Bcl-2	上游：GGTGGGGTCATGTGTGTGG	197
Bcl-2	下游：CGGTTCAGGTACTCAGTCATCC	197
β-actin	上游：TCAGGTCATCACTATCGGCAAT	134
β-actin	下游：AAAGAAAGGGTGTAAAACGCA	134

组别	HDL-C	总胆固醇	TG
对照组	1.75±0.18	0.33±0.04	1.05±0.11
SANFH组	0.58±0.06^a	1.11±0.12^a	2.65±0.26^a
TFRD-低组	0.85±0.09^b	0.78±0.08^b	2.05±0.20^b
TFRD-中组	1.22±0.13^bc	0.52±0.06^bc	1.42±0.14^bc
TFRD-高组	1.71±0.18^bcd	0.35±0.04^bcd	1.08±0.11^bcd
TFRD-高+抑制剂组	0.88±0.09^e	0.81±0.09^e	1.95±0.20^e
F	137.728^＊	155.765^＊	123.687^＊

组别	HDL-C	总胆固醇	TG
对照组	1.75±0.18	0.33±0.04	1.05±0.11
SANFH组	0.58±0.06^a	1.11±0.12^a	2.65±0.26^a
TFRD-低组	0.85±0.09^b	0.78±0.08^b	2.05±0.20^b
TFRD-中组	1.22±0.13^bc	0.52±0.06^bc	1.42±0.14^bc
TFRD-高组	1.71±0.18^bcd	0.35±0.04^bcd	1.08±0.11^bcd
TFRD-高+抑制剂组	0.88±0.09^e	0.81±0.09^e	1.95±0.20^e
F	137.728^＊	155.765^＊	123.687^＊

组别	BV/TV/%	Tb.N/mm^-1
对照组	45.05±4.53	2.64±0.28
SANFH组	17.22±1.79^a	0.98±0.10^a
TFRD-低组	25.31±2.61^b	1.35±0.14^b
TFRD-中组	33.62±3.42^bc	1.86±0.19^bc
TFRD-高组	44.23±4.51^bcd	2.54±0.26^bcd
TFRD-高+抑制剂组	25.44±2.61^e	1.41±0.15^e
F	108.701^＊	117.015^＊