RNA-Seq revealed the effect of adding different proportions of wheat diet on fat metabolism of Tibetan lamb.

The objective of this study was to evaluate the effect of the level of wheat substitution for corn on the fat metabolism of Tibetan lamb. A total of 90 Tibetan lambs [body weight (BW) of 19.78 ± 2.45 kg] were arranged with three substitution levels of wheat: WC (100 % corn), WL (10 % wheat substitution for corn), and WH (15 % wheat substitution for corn) on a dry matter basis. After the experiment, cry section technology was used to scrutinize the subcutaneous adipose tissue morphology, and genes related to fat metabolism were excavated using high-throughput sequencing technology. According to the study results, fat diameter and fat biovolume of the WL and WH groups were less than the WC group. A total of 506 differentially expressed genes (DEGs) were identified by RNA sequencing (RNA-Seq) technology. Compared with the WC group, 66 DEGs were upregulated and 59 DEGs were downregulated in the WL group, and 179 DEGs were upregulated and 269 DEGs were downregulated in the WH group. The top 20 DEGs were analyzed by Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway, indicating significant differences in the fat metabolism pathway. Five DEGs were randomly screened for quantitative reverse transcription-polymerase chain reaction (qRT-PCR) verification, and the results were consistent with the RNA-Seq results, which proved the accuracy of sequencing. In summary, with the increase in the proportion of supplemental wheat, the fat cells became smaller, and the genes related to fat decomposition were significantly upregulated.

[Mechanism of Huanglian Wendan Decoction in improving impaired glucose tolerance based on skeletal muscle NLRP3/caspase-1/IL-1ß, IL-18 pathway].

This study investigated the mechanism of improving impaired glucose tolerance(IGT) of rats by Huanglian Wendan Decoction from the perspective of the skeletal muscle Nod-like receptor protein 3(NLRP3)/cysteinyl aspartate specific proteinase-1(caspase-1)/interleukin-1ß(IL-1ß), interleukin-18(IL-18) pathway. Healthy male SD rats were fed with the diet containing 45% fat for 20 weeks, accompanied by a high-temperature and high-humidity environment and an inactive lifestyle, for the establishment of the IGT rat model. The rats were divided into the blank control group, model control group, metformin hydrochloride group(positive drug group, 0.05 g·kg~(-1)·d~(-1)) and Huanglian Wendan Decoction group(7.8 g·kg~(-1)·d~(-1)). After continuous intragastric administration for 4 weeks, the obesity and glycemic indexes of all the rats were measured. The fasting serum insulin(FINS) level was determined by ELISA, with the insulin sensitivity index(ISI) and insulin resistance index(IRI) calculated. The mRNA and protein expression le-vels of nuclear factor kappaB(NF-κB), NLRP3, caspase-1, IL-1ß and IL-18 in skeletal muscle tissue were detected by real-time polymerase chain reaction(PCR), Western blot and immunofluorescence. Compared with the blank control group, the model control group witnessed significantly increased mRNA and protein expression of NF-κB, NLRP3, caspase-1, IL-1ß and IL-18. As revealed by the comparison with the model control group, Huanglian Wendan Decoction could effectively regulate the obesity status, reduce body weight, correct the abnormal levels of 2-hour plasma glucose(2 hPG), insulin resistance index(IRI), insulin sensitivity index(ISI), and lower the mRNA and protein expression of NF-κB, NLRP3, caspase-1, IL-1ß and IL-18 in the skeletal muscle tissue of IGT rats. Combined with previous studies, the above results showed that the occurrence and development of IGT was closely related to inflammatory response and the classic pyroptosis pathway in skeletal muscle, such as NLRP3/caspase-1/IL-1ß, IL-18. It is inferred that the mechanism of Huanglian Wendan Decoction was to alleviate insulin resistance(IR) and then reverse the course of IGT lies in the regulation of the abnormal insulin receptor signaling pathway based on the NLRP3 inflammasome pathway.

[Mechanism of Huanglian Wendan Decoction in intervening IGT IR based on liver cell pyroptosis].

To investigate the mechanism of Huanglian Wendan Decoction in intervening impaired glucose tolerance( IGT) rat insulin resistance( IR) based on the pyroptosis pathway of liver cells. The IGT rat model was established by high-fat diet( 20 weeks) combined with high temperature,humidity environment and inactive lifestyle. The experiment was divided into blank group,model group,metformin hydrochloride group( positive group)( 0. 05 g·kg~(-1)·d~(-1)) and Huanglian Wendan Decoction group( 7. 8 g·kg~(-1)·d~(-1)). After continuous intragastric administration for 4 weeks,the body weight,body length and abdominal circumferences of all the rats were measured,the Lee' s obesity indexes was calculated,and the levels of fasting plasma glucose( FPG) and 2-hour plasma glucose( 2 hPG) in each group were measured. Serum insulin levels( FINS) and tumour necrosis factor-α( TNF-α) levels were detected by ELISA,and insulin sensitivity index( ISI) and insulin resistance index( IRI) values were calculated. The expressions of cysteinyl aspartate specific proteinase-1( caspase-1),gasdermin D( GSDMD),interleukin-1ß( IL-1ß) and interleukin-18( IL-18) m RNA and proteins in liver tissues were detected by Real-time PCR and Western blot. Immunofluorescence was used to detect the expressions of caspase-1 and GSDMD protein in liver tissues. Huanglian Wendan Decoction could effectively reduce the weight of model rats,improve abdominal obesity,FINS,IRI and ISI indexes and correct 2 hPG levels. Compared with blank group,TNF-α levels in serum and caspase-1,GSDMD,IL-1ß and IL-18 expressions in liver tissue of model control group were increased significantly. Huanglian Wendan Decoction could effectively reduce TNF-α level in serum,regulate the expressions of caspase-1,GSDMD,IL-1ß and IL-18 genes and proteins in liver tissues of IGT rats. The above results showed that the occurrence and development of " obesity-IR-abnormal glucose metabolism-type 2 diabetes mellitus" was closely related to inflammatory response and the classical pyroptosis pathway mediated by caspase-1/GSDMD. The mechanism of Huanglian Wendan Decoction in improving IR may be correlated with reduction of the level of inflammatory factors and the alleviation of the state of pyroptosis,and thus reverse the course of IGT.