1.Analysis of Mechanism of Qinggan Jianpi Huoxue Prescription in Treatment of Hepatic Fibrosis Rats by Regulating M1/M2 Macrophages
Fuzhen PAN ; Hongxin CAO ; Yongsheng ZHANG ; Xiaqiu WU ; Weifang ZHENG ; Ding LIU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(21):94-102
ObjectiveTo observe the effect of Qinggan Jianpi Huoxue prescription(QGJPHXP) on the polarization of M1/M2 macrophages in rats with hepatic fibrosis induced by carbon tetrachloride(CCl4). MethodA rat hepatic fibrosis model was established by intraperitoneal injection of 40% CCl4-olive oil suspension twice a week at the dosage of 2.0 mL·kg-1 for 8 weeks. After the model was successfully established, these rats were randomly divided into the model group, QGJPHXP group(32.084 g·kg-1) and Biejiajian pills(BJJP) group(0.925 5 g·kg-1), with 12 rats in each group. The blank group was injected intraperitoneally with the same amount of olive oil. The rats in the administration groups were given the corresponding solution according to the dose, and the blank and model groups were given the same dose of purified water, once a day. After 4 weeks of continuous administration, the liver tissues of rats were taken and stained with hematoxylin-eosin(HE) and Masson to observe the pathological changes. The serums were collected to detect the alanine aminotransferase(ALT) and aspartate aminotransferase(AST) levels. Interleukin(IL)-6, IL-12, IL-10, IL-1β, transforming growth factor-β1(TGF-β1) and tumor necrosis factor-α(TNF-α) levels in liver tissues were measured by enzyme-linked immunosorbent assay(ELISA). The expression levels of CD86 and CD206 were detected by immunohistochemistry(IHC). Western blot and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) were used to detect the protein and mRNA expression levels of inducible nitric oxide synthase(iNOS), arginase-1(Arg-1), phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK), nuclear transcription factor-κB p65(NF-κB p65) in liver tissues of rats. ResultCompared with the blank group, the hepatic cell plate was irregularly arranged, and local inflammatory cell infiltration and fibrous hyperplasia were observed, while the serum levels of ALT and AST were significantly increased in the model group(P<0.01), and IL-1β, IL-6, IL-12, TGF-β1, TNF-α, CD86, CD206, iNOS, p-p38 MAPK,p38 MAPK and NF-κB p65 levels in liver tissues were obviously increased(P<0.05, P<0.01), while the levels of IL-10 and Arg-1 were obviously decreased(P<0.05, P<0.01). Compared with the model group, QGJPHXP group reduced the degree of liver cell fibrosis,and serum levels of ALT and AST(P<0.01), and IL-1β, IL-6, IL-12, TGF-β1, TNF-α, CD86, iNOS, p-p38 MAPK, p38 MAPK, and NF-κB p65 levels in liver tissues were obviously decreased(P<0.05, P<0.01), the levels of IL-10, CD206 and Arg-1 were obviously increased in the QGJPHXP group(P<0.05, P<0.01). ConclusionQGJPHXP has ability to inhibit the activation of pro-inflammatory M1 macrophages, induce the secretion of anti-inflammatory cytokines by M2 macrophages, reduce the release of pro-fibrogenic cytokines, and promote the macrophage polarization of M1 to M2 in liver for tissue repair, thereby serving as an anti-inflammatory and anti-hepatic fibrosis drug.