Mitochondrial dysfunction in a rat model and the related risk of metabolic disorders

doi:10.19852/j.cnki.jtcm.20221017.001

Abstract

Abstract:

OBJECTIVE: To explore whether kidney Yang deficiency (KYD) is prone to metabolic disorders may be linked to impaired mitochondrial function in thermogenesis and metabolic tissues.
METHODS: A rat model of KYD was used, which was established using Sprague Dawley rat dams with warm preference subjected to herbal treatment that can improve kidney Yang. The human relevance was confirmed by reduced serum corticosterone levels, and increased preference for warm location.
RESULTS: KYD Rats were underdeveloped. Adenosine-triphosphate (ATP) production was reduced in the brown fat, but increased in the muscle. However, oxidative phosphorylated complexes to generate ATP and mitochondrial biogenesis marker were reduced in both tissues. When the second insult of high-fat diet (HFD) was introduced, KYD rats gained less weight yet developed more severe lipid and glucose metabolic disorders. This may be driven by disregulated liver gluconeogenesis marker forkhead box protein O1 and lipid metabolic regulator cholesterol 7 alpha-hydroxylase.
CONCLUSION: KYD rats exhibited reduced mito-chondrial function in the brown fat, but were partially compensated by skeletal muscle, associated with the phenotype of warm preference and metabolic disorder, which was further exacerbated by additional HFD consumption. Future studies can focus on treatment targetting mitochondria function to reverse this phenotype.

Key words: kidney Yang deficiency, DNA, mitochondrial, adenosine triphosphate, thermogenesis, peroxisome proliferator-activated receptor gamma coactivator 1-alpha

LI Han, HUANG Xiaomin, CAI Haiyang, HEROK George, HE Jing, SU Yixun, LI Weihong, YI Chenju, OLIVER Brian G, CHEN Hui. Mitochondrial dysfunction in a rat model and the related risk of metabolic disorders[J]. Journal of Traditional Chinese Medicine, 2023, 43(1): 95-104.

Figures/Tables 6

Table 1 Biometric parameters and blood metabolic markers in male offspring at 13 weeks

Parameter	Control (n = 10)	KYD (n = 10)
Birth weight at birth (g)	7.800±0.200	4.780±0.287^b
Body weight at 13 weeks (g)	356.850±6.134	325.200±11.404^a
Liver (g)	10.285±0.270	10.680±0.236
Liver as a percentage of body weight (%)	2.883±0.063	3.323±0.140^a
Kidney (g)	2.456±0.100	2.059±0.065^a
Kidney as a percentage of body weight (%)	0.705±0.036	0.643±0.036
Brown adipose tissue (g)	0.497±0.051	0.465±0.054
Brown adipose tissue as a percentage of body weight (%)	0.139±0.015	0.147±0.021
White adipose tissue (g)	4.961±0.354	2.800±0.310^b
White adipose tissue as a percentage of body weight (%)	1.391±0.098	0.856±0.082^b
Sketeal muscle (g)	0.534±0.047	0.694±0.044^a
Sketeal muscle as a percentage of body weight (%)	0.152±0.014	0.218±0.019^a
Serum corticosterone (ng/mL)	22.264±0.771	15.190±1.415^a
Serum T3 (ng/mL)	3.841±0.152	3.675±0.113
Blood glucose (mM)	8.050±0.371	7.825±0.508
Serum LDL (mM)	1.105±0.062	1.257±0.027^a
Serum cholesterol (mM)	2.210±0.081	2.763±0.121^b

Figure 1 Markers involved in temperature and thermogenesis A: hot plate test; B: body temperature; C: mRNA expression of UCP1 in the BAT; D: mRNA expression of UCP1 in the skeletal muscle. KYD: kidney Yang deficiency; UCP1: uncoupling protein 1; BAT: brown adipose tissue. Results are expressed as mean ± standard error of the mean, n = 6-10. aP < 0.05, vs control.

Figure 2 Mitochondrial metabolic markers in the BAT A: ATP levels; B; peroxisome PCG1α mRNA expression; C: mitoDNA copy number; D: oxidative phosphorylation complexes CI; E: CV in the BAT in male offspring at 13 weeks; F: total and mitochondrial protein from brown adipose tissue. BAT: brown adipose tissue; PCG1α: peroxisome proliferator-activated receptor gamma coactivator 1-alpha; mitoDNA: mitochondrial DNA; OXPHOS CV: the complexes V of the oxidative phosphorylation system; OXPHOS CI: the complexes I of the oxidative phosphorylation system; KYD: kidney Yang deficiency; ATP: adenosine-triphosphate; Results are expressed as mean ± standard error of the mean, n = 4-6. aP < 0.05, bP < 0.01, cP < 0.001, vs control.

Figure 3 Mitochondrial metabolic markers in the muscle A: ATP levels; B: PCG1α mRNA expression; C: mitoDNA copy number; D: oxidative phosphorylation complexes CI and, E: CV in the skeletal muscle in male offspring at 13 weeks; F: total and mitochondrial protein was extracted from skeletal muscle. Control: normal SD mouse. ATP: adenosine-triphosphate; PCG1α: peroxisome proliferator-activated receptor gamma coactivator 1-alpha; mitoDNA: mitochondrial DNA; OXPHOS CV: the complexes V of the oxidative phosphorylation system; OXPHOS CI: the complexes I of the oxidative phosphorylation system; KYD: kidney Yang deficiency. Results are expressed as mean ± standard error of the mean, n = 4-6. aP < 0.05, bP < 0.01, vs Control.

Table 2 Biometric parameters and blood metabolic markers in male offspring fed a HFD

Item	Control-chow (n = 6)	KYD-chow (n = 6)	Control-HFD (n = 6)	KYD-HFD (n = 6)
Body weight before the diet (g)	53.000 ± 0.512	50.833 ± 1.426	52.000 ± 1.348	53.000 ± 0.688
Body weight at endpoint (g)	375.000 ± 9.000	323.000 ± 15.000^a	442.000 ± 12.000^b	380.000 ± 9.000^df
Caloric intake (kJ/d per rat)	271.000 ± 16.000	275.000 ± 14.000	348.000 ± 12.000^a	403.000 ± 35.000^f
Liver (g)	11.787 ± 0.415	10.721 ± 0.891	13.657 ± 0.907	14.203 ± 0.784^f
Liver as a percentage of body weight (%)	2.965 ± 0.202	3.011 ± 0.071	3.163 ± 0.152	3.712 ± 0.090^f
White adipose tissue (g)	4.993 ± 0.822	3.562 ± 0.571	9.758 ± 1.710^a	8.465 ± 0.904^e
White adipose tissue as a percentage of body weight (%)	1.275 ± 0.203	0.894 ± 0.080	1.883 ± 0.222 ^a	2.200 ± 0.203^f
Sketeal muscle (g)	0.717 ± 0.043	0.744 ± 0.057	0.824 ± 0.035	0.781 ± 0.024
Sketeal muscle as a percentage of body weight (%)	0.184 ± 0.010	0.199 ± 0.003	0.200 ± 0.009	0.202 ± 0.007
Blood insulin (ng/mL)	0.147 ± 0.026	0.124 ± 0.013	0.291 ± 0.046^b	0.154± 0.031^d
Serum LDL (mM)	1.199 ± 0.067	1.414 ± 0.054^a	1.489 ± 0.075^b	2.008 ± 0.080^cf
Serum Cholesterol (mM)	2.514 ± 0.050	2.712 ± 0.080	3.000 ± 0.184^a		3.515 ± 0.173^ce

Figure 4 Energy metabolic regulators in the muscle and liver A: PCG1α mRNA expression; B: PPARγ mRNA expression; C: ratios between (IRS1 and total IRS1; D: ratios between between Akt and total Akt in the sketela muscle; E: mRNA expression of FOXO1; F: mRNA expression of CYP7A1 in the liver in chow and HFD-fed rats at 13 weeks. PCG1α: peroxisome proliferator-activated receptor gamma coactivator 1-alpha; PPARγ: peroxisome proliferator-activated receptor gamma; IRS1: p-insulin receptor substrate 1; Akt: p-protien kinase B; FOXO: forkhead box protein O; CYP7A1: cholesterol 7 alpha-hydroxylase; KYD: kidney Yang deficiency; HFD: high-fat diet; ATP: adenosine-triphosphate. Results are expressed as mean ± standard error of the mean, n = 6. aP < 0.05, bP < 0.01, vs Control-chow; cP < 0.01 vs KYD-chow; dP < 0.01vs Control-HFD.

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