Effect and Mechanism of Cordyceps militaris Extract on Lowering Uric Acid in Hyperuricemia Rats

doi:10.13560/j.cnki.biotech.bull.1985.2024-0379

Abstract

Abstract:

【Objective】 To investigate the effect and the corresponding mechanism of Cordyceps militaris（C. militaris）extract on lowering uric acid by hyperuricemia model rats. 【Method】 The contents of total sugars, reducing sugars, polysaccharides, proteins and polyphenols in C. militaris extract were determined. The rat model of hyperuricemia was established based on feeding of potassium oxonate combined with yeast paste. The serum uric acid level, renal function, oxidative stress and inflammation of rats and the changes of intestinal microbial composition were measured to evaluate the effect of C. militaris extract on hyperuricemia rats and its mechanism. 【Result】 The C. militaris extract contained 35.86% polysaccharides, 27.05% proteins, and 0.21% polyphenols. Animal experiments demonstrated that C. militaris extract significantly reduced the serum uric acid levels in hyperuricemia rats. The most effective dose was found to be 0.5 g/(kg·d), leading to a reduction in uric acid levels from 281.62 μmol/L to 93.27 μmol/L, with an inhibition rate of 66.88%. It also inhibited the oxidative stress and inflammation in renal tissue of hyperuricemia rats. Molecular mechanism studies showed that C. militaris extract down-regulated uric acid transporter1（URAT1）and glucose transporter 9（GLUT9）, up-regulated the expressions of organic anion transporter 1（OAT1）and ATP binding cassette subfamily G member 2（ABCG2）in renal tissues of hyperuricemia rats. Concurrently, C. militaris extract significantly inhibited the xanthine oxidase activity in liver tissues. In addition, C. militaris extract enhanced the diversity of intestinal microbiota and maintained the balance of intestinal microecology in hyperuricemia rats. 【Conclusion】 C. militaris extract demonstrated a good anti-hyperuricemia effect, which might be related to regulating the expression of uric acid transporter and inhibiting the activity of xanthine oxidase.

Key words: hyperuricemia, Cordyceps militaris, water extract, uric acid transporters, gut microbiota

XIONG Xin-yi, LIU Li-ping, FENG Jie, ZHANG Jin-song, LI De-shun, LIU Peng, LIU Yan-fang. Effect and Mechanism of Cordyceps militaris Extract on Lowering Uric Acid in Hyperuricemia Rats[J]. Biotechnology Bulletin, 2024, 40(11): 34-46.

Figures/Tables 11

Table 1 Primers for RT-qPCR target genes

基因Gene	引物序列Primer sequence（5'-3'）
URAT1	F: GCTACCAGAATCGGCACGCT
URAT1	R: CACCGGGAAGTCCACAATCC
GLUT9	F: GAGATGCTCATTGTGGGACG
GLUT9	R: GTGCTACTTCGTCCTCGGT
ABCG2	F: GTAGGTCGGTGTGCGAGTCA
ABCG2	R: AACCAGTTGTGGGCTCATCC
OAT1	F: GGCACCTTGATTGGCTATGT
OAT1	R: CCACAGCATGGAGAGACAGA
TNF-α	F: GCGTGTTCATCCGTTCTCTACC
TNF-α	R: TACTTCAGCGTCTCGTGTGTTTCT
IL-6	F: AGTTGCCTTCTTGGGACTGATGT
IL-6	R: GGTCTGTTGTGGGTGGTATCCTC
β-actin	F: GCAGGAGTACGATGAGTCCG
β-actin	R: ACGCAGCTCAGTAACAGTCC

Table 2 Analysis of amino acids and nucleosides components in C. militaris extract

氨基酸成分 Amino acid component	含量 Content/%	氨基酸成分 Amino acid component	含量 Content/%	核苷类成分 Nucleosides component	含量 Content/%
L-组氨酸	0.16	L-脯氨酸	0.41	胞嘧啶	0.01
L-精氨酸	0.94	L-鸟氨酸	2.31	胞苷	0.03
L-天冬酰胺	0.25	D-2-氨基丁酸	0.02	鸟嘌呤	0.02
L-谷氨酰胺	0.01	L-赖氨酸	1.08	腺嘌呤	0.03
L-丝氨酸	0.59	L-酪氨酸	0.34	尿苷	0.41
L-甘氨酸	0.14	L-甲硫氨酸	0.09	胸腺嘧啶	0.04
L-天冬氨酸	0.68	L-缬氨酸	0.64	肌苷	0.06
L-瓜氨酸	0.02	L-异亮氨酸	0.34	鸟苷	0.21
L-谷氨酸	1.92	L-亮氨酸	0.65	胸苷	0.02
L-苏氨酸	0.76	L-苯丙氨酸	0.28	腺苷	0.34
L-丙氨酸	1.57	L-色氨酸	0.10	虫草素	0.83
γ-氨基丁酸	0.25			N⁶-（2-羟乙基）腺苷	0.54

Fig. 1 FT-IR spectrum of C. militaris extract

Fig. 2 Effects of C. militaris extract on renal function and renal pathological injury in hyperuricemia rats A: Serum uric acid level; B: serum urea nitrogen concentration; C: serum creatinine concentration; D: renal fibrotic area; E: HE staining of kidney; F: masson staining of kidney. Compared with CTL group, ###: P<0.001；compared with MC group, **: P<0.01, ***: P<0.001. Scale bar: 100 μm

Fig. 3 Effects of C. militaris extract on renal oxidative stress in hyperuricemia rats A: Renal GSH content; B: renal MDA content; C: renal T-AOC. Compared with CTL group, #: P<0.05；compared with MC group, **: P<0.01, ***: P<0.001

Fig. 4 Effects of C. militaris extract on renal inflammation in hyperuricemia rats A: IHC staining of CD68; B: IHC staining of NLRP3; C: IHC staining of IL-1β; D: positive area of CD68; E: relative mRNA levels of IL-6; F: relative mRNA levels of TNF-α; G: positive area of NLRP3; H: positive area of IL-1β. Compared with CTL group, ###: P<0.001；compared with MC group, **: P<0.01, ***: P<0.001. Scale bar: 100 μm

Fig. 5 Effects of C. militaris extract on renal uric acid transporter in hyperuricemia rats A: IHC staining of URAT1; B: IHC staining of OAT1; C: positive area of URAT1; D: relative mRNA levels of URAT1; E: relative mRNA levels of GLUT9; F: positive area of OAT1; G: relative mRNA levels of OAT1; H: relative mRNA levels of ABCG2. Compared with CTL group, ###: P<0.001；compared with MC group, **: P<0.01, ***: P<0.001. Scale bar: 100 μm

Fig. 6 Effects of C. militaris extract on XOD activity and liver pathological injury in hyperuricemia rats A: XOD activity in kidney; B: HE staining of kidney. Compared with CTL group, ###: P<0.001；compared with MC group, *: P<0.05, **: P<0.01, ***: P<0.001. Scale bar: 100 μm

Fig. 7 PCA analysis（A）and LEfSe multilevel species-level tree（B）of gut microbiota in hyperuricemia rats

Fig. 8 Effects of C. militaris extract on the relative abundances of key gut microbiota in rats with hyperuricemia

Fig. 9 Correlation analysis of differential OTUs and biochemical indexes

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