Found programs: Natural Science Research Project of Anhui Educational Committee (No.2022AH050651); Scientific Research Project of Anhui Medical University (No. 2022xkj107); Natural Science Key Project of Bengbu Medical College (No.2022byzd196)
Authors：Bai Hongmei1, Yang Zhen1, Hu Weikang1 , Wang Zihan1, Zhou Wenjing1, He Qingya1, Zhong Jian1, Li Mingcong2, Liu Li3, Zhang Chaoyang3, Zhang Sumei1, Zhang Shengquan1
Keywords：lanosterol synthetase; cholesterol metabolism; non-alcoholic fatty liver disease;intestinal permeability; tight junction protein; intestinal barrier function
DOI：专辑：医药卫生科技
〔Abstract〕 Abstract Objective To investigate the effect of loss of function of lanosterol synthase(LSS) on intestinal function in a mouse model of non-alcoholic fatty liver disease(NAFLD) induced by a high-fat diet. Methods LSS heterozygous knockout C57 mice (LSS+/-) were established using the CRISPR/CAS9 system. After being fed a high-fat diet with 60% fat content for 6 months, the fat deposition in liver tissues was detected by HE and Oil red O staining, the morphological changes of small intestine tissue were detected by HE staining. The changes in total cholesterol content in intestinal tissue were detected by kits. The gastrointestinal motility function of mice was detected by phenol red paste. The intestinal permeability was detected by Evans blue staining, and the expression of lanosterol synthase (LSS), tight junction protein (Claudin)-1, Claudin-5, platelet glycoprotein IV (CD36), and Niemann-Pick C1-like 1 (NPC1L1) proteins in small intestinal tissues were detected by Western blot. Results The results of HE and Oil red O staining of liver tissues showed that liver fat deposition in LSS knockdown mice was lower than that in wild-type mice in the high-fat diet group. The total cholesterol content in intestinal tissue of LSS gene knockout mice decreased (P<0.01), but no morphological differences were observed between the two groups of mice by HE staining of intestinal tissues. The gastrointestinal motility function of LSS gene knockout mice did not show significant changes. The intestinal permeability of LSS gene knockout mice in the high-fat diet group decreased as detected by Evans blue (P < 0.05). The expression levels of Claudin-5 protein in the intestinal tissue of LSS gene knockout mice in the high-fat diet group increased (P<0.05), while the expression of LSS in the intestinal tissues of LSS knockout mice decreased (P<0.05). Conclusion In the NAFLD model induced by a high-fat diet,LSS gene knockout reduces liver fat deposition induced by a high-fat diet and improves intestinal barrier function by regulating cholesterol metabolism in intestinal tissues and up-regulating the expression of tight junction protein-5.