Qian Jun, Jiao Yuwen, Wang Guangyao, Liu Hanyang, Cao Xiang, Yang Haojun
Department of General Surgery, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou, Jiangsu 213000, P.R. China.
Exp Ther Med. 2020 Aug;20(2):1541-1549. doi: 10.3892/etm.2020.8826. Epub 2020 May 30.
Effect of exogenous transforming growth factor-β1 (TGF-β1) on cholestatic mice by inhibiting Kupffer cell immune responses in liver was investigated. To induce cholestasis, BALB/c mice received a sham operation (Mock group), or underwent a bile duct ligation (BDL group) and then were subcutaneously injected with TGF-β1 at multiple sites (TGF group). Liver functions were evaluated according to the levels of alanine aminotransferase (ALT), aspartate aminotransferase AST and γ-glutamyltranspeptidase (γ-GT) in serum samples. Expression of nuclear factor-κB (NF-κB), interleukin-6 (IL-6), IL-1β and tumor necrosis factor-α (TNF-α) was detected. Expression of inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1) in Kupffer cells (KCs) of the liver was detected. The isolated KCs were divided into control group, LPS group, TGF group and Galunisertib group and western blot analysis was used to detect the expression of NF-κB, IL-6, IL-1β, TNF-α, iNOS and Arg-1. The percentage of CD40, CD86, CD204 and CD206 as macrophage cell surface antigens were measured by flow cytometry. The indexes of liver function and liver fibrosis of the mice in the TGF group were significantly lower than those in the BDL group (P<0.05). The levels of IL-1β, IL-6 and TNF-α in the liver were lower than those in the BDL group, while the level of IL-10 was significantly increased (P<0.05). M2-type transformation occurred in liver Kupffer cells of mice in the TGF group. In cell experiments, TGF treatment downregulated the expression of IL-1β, IL-6, TNF-α and NF-κB, increased the expression of IL-10, and induced M2-type transformation in macrophages (P<0.05). In conclusion, TGF-ß1 diminished the progression of cholestasis in mice by inhibiting the inflammatory response of KCs and regulating KC polarization.
研究了外源性转化生长因子-β1(TGF-β1)通过抑制肝脏库普弗细胞免疫反应对胆汁淤积小鼠的影响。为诱导胆汁淤积,BALB/c小鼠接受假手术(假手术组),或进行胆管结扎(胆管结扎组),然后在多个部位皮下注射TGF-β1(TGF组)。根据血清样本中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和γ-谷氨酰转肽酶(γ-GT)的水平评估肝功能。检测核因子-κB(NF-κB)、白细胞介素-6(IL-6)、IL-1β和肿瘤坏死因子-α(TNF-α)的表达。检测肝脏库普弗细胞(KCs)中诱导型一氧化氮合酶(iNOS)和精氨酸酶-1(Arg-1)的表达。将分离的KCs分为对照组、LPS组、TGF组和加鲁尼西替布组,采用蛋白质免疫印迹分析检测NF-κB、IL-6、IL-1β、TNF-α、iNOS和Arg-1的表达。通过流式细胞术检测巨噬细胞表面抗原CD40、CD86、CD204和CD206的百分比。TGF组小鼠的肝功能和肝纤维化指标显著低于胆管结扎组(P<0.05)。肝脏中IL-1β、IL-6和TNF-α的水平低于胆管结扎组,而IL-10水平显著升高(P<0.05)。TGF组小鼠肝脏库普弗细胞发生M2型转化。在细胞实验中,TGF处理下调了IL-1β、IL-6、TNF-α和NF-κB的表达,增加了IL-10的表达,并诱导巨噬细胞发生M2型转化(P<0.05)。总之,TGF-β1通过抑制KCs的炎症反应和调节KC极化减轻了小鼠胆汁淤积的进展。