牦牛骨骼肌中特异基质金属蛋白酶及其功能相关基因的表达分析

doi:10.16431/j.cnki.1671-7236.2023.05.005

Abstract

Abstract: 【Objective】 This study was aimed to screen and identify the members of matrix metalloproteinases (MMPs) family and their functional related genes specifically expressed in skeletal muscle of yak,in order to provide a basis for studying the formation mechanism of skeletal muscle structure in yak.【Method】 Glutei and pectoral muscle of adult male yaks were collected.The characteristics of muscle fibers and the number of collagen fibers were analyzed by histochemical staining (HE and Masson).The mRNA expression of MMPs and matrix metalloproteinase inhibitors (TIMPs) were detected by Real-time quantitative PCR.Western blotting and immunohistochemistry were used to analyze the protein expression and tissue distribution of MMPs,types Ⅰ and Ⅲ collagen.Protein-protein interaction networks analysis of MMPs and their functionally related genes was constructed by STRING website.EnzChek^TM Gelatinase/Collagenase kit was used to detect the differences in enzyme activities of gelatinase/collagenase between gluteus and pectoral muscles.【Result】 Masson staining and Real-time quantitative PCR analysis results showed that the area of collagen fiber,the expression of MMP-2 and MMP-14 genes in gluteus of yak was significantly lower than that in pectoral muscle (P＜0.05),whereas the expression of TIMP-3 gene was the opposite (P＜0.05).Western blotting results showed that the expression of MMP-3,MMP-14,types Ⅰ and Ⅲ collagen in gluteus were extremely significantly or significantly lower than those in pectoral muscle (P＜0.01 or P＜0.05).Immunohistochemical analysis results showed that the expression of MMP-2,MMP-9,MMP-14 and type Ⅰ collagen in gluteus were significantly or extremely significantly lower than that in pectoral muscle (P＜0.05 or P＜0.01),but the expression of MMP-16 was the opposite (P＜0.05).Protein function correlation clustering analysis showed that MMP-2,MMP-14,MMP-16,TIMP-2,TIMP-3,COL1A2 and COL3A1 could cluster together,and MMP-3,MMP-9 and TIMP-1 could cluster together.The enzyme activities of gelatinase/collagenase in gluteus and pectoral muscle showed a similar tendency,and the difference was not significant (P＞0.05).【Conclusion】 Members of the MMPs family and their functionally related genes were widely expressed in skeletal muscle of yak,and the expression were different between gluteus and pectoral muscle,which provided reference basis for further study on the mechanism of MMPs regulating the structure formation of skeletal muscle in yak.

Key words: yak; skeletal muscle; matrix metalloproteinases (MMPs); collagen

CLC Number:

Q786

WU Zhijuan, CHAI Zhixin, WANG Jikun, WANG Jiabo, ZHONG Jincheng, XIN Jinwei. Expression Analysis of Specific Matrix Metalloproteinases and Functionally Related Genes of Skeletal Muscles in Yak[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1774-1784.

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Expression Analysis of Specific Matrix Metalloproteinases and Functionally Related Genes of Skeletal Muscles in Yak

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