Foreign Language Department, Guangxi University of Chinese Medicine, Nanning, China.
Department of Liver Diseases, The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning, China.
Biomed Res Int. 2022 May 11;2022:4479885. doi: 10.1155/2022/4479885. eCollection 2022.
To research the influence of Chinese medicine Jiedu Huayu granules (JDHY) on the immune response and inflammatory response of rats with acute liver failure (ALF) and investigate its related mechanism.
Rats were randomly divided into 4 groups: control group ( = 6) were injected with the same amount of normal saline; ALF group ( = 10) were injected intraperitoneally with D-GaIN (700 mg/kg) and LPS (10 g/kg); ALF+JDHY group ( = 10) were given JDHY 57.55 g/kg/d by gavage for 7 days and injected intraperitoneally with D-GaIN/LPS after the last dose; and ALF+BAY group ( = 10) were given BAY 10 mg/kg/d by gavage for 7 days and injected intraperitoneally with D-GaIN/LPS after the last dose. Changes in liver function and coagulation function were examined in rat serum; the pathological varieties of liver tissues were verified by HE staining; immunohistochemistry was utilized to determine the ratio of PCNA and F4/80 in liver tissues; the flow cytometry was applied to determine the ratio of CD4+/CD8+ cells in peripheral blood mononuclear cells (PBMCs); ELISA and qRT-PCR were utilized to check the level of IL-10, IL-6, IL-13, IL-1, TNF-, IFN-, and CD163 in serum and liver cells. Western blot was adopted to check the expression of apoptotic protein and expression and NF-B pathway-related protein expression.
JDHY and BAY could decline the expression of AST, ALT, ALP, and TBiL in ALF rat serum significantly ( < 0.01), increase PTA and PLT ( < 0.01), and mitigate liver tissue damage. Besides, JDHY and BAY could reduce the apoptosis and improve the proliferation of the liver cells in rats with ALF; meanwhile, the ratio of CD4+ cells and F4/80 cells was reduced while CD8+ cells were increased ( < 0.01). Further, JDHY and BAY could reduce the level of IFN-, IL-6, IL-1, and TNF- while increasing the level of IL-10 and IL-13 ( < 0.01). Additionally, the expression of sCD163 in serum and CD163 expression in liver tissues increased ( < 0.01). The result of western blot confirmed that JDHY could inhibit the phosphorylated expression of NF-B, IK, and IKK in the ALF rat tissues.
JDHY can upregulate the level of CD163/sCD163 by the NF-B signaling pathway, thereby regulating immune response, inhibiting inflammatory response, and ultimately improving ALF in the rats.
研究中药解毒化瘀颗粒(JDHY)对急性肝衰竭(ALF)大鼠免疫应答和炎症反应的影响,并探讨其相关机制。
将大鼠随机分为 4 组:对照组(n = 6)注射等体积生理盐水;ALF 组(n = 10)腹腔注射 D-GaIN(700 mg/kg)和 LPS(10 g/kg);ALF+JDHY 组(n = 10)给予 JDHY 57.55 g/kg/d 灌胃 7 天,末次给药后腹腔注射 D-GaIN/LPS;ALF+BAY 组(n = 10)给予 BAY 10 mg/kg/d 灌胃 7 天,末次给药后腹腔注射 D-GaIN/LPS。检测大鼠血清肝功能和凝血功能变化;HE 染色验证肝组织病理变化;免疫组化法检测肝组织 PCNA 和 F4/80 比值;流式细胞术检测外周血单个核细胞(PBMCs)中 CD4+/CD8+细胞比值;ELISA 和 qRT-PCR 检测血清和肝组织中 IL-10、IL-6、IL-13、IL-1、TNF-、IFN-和 CD163 水平。Western blot 检测凋亡蛋白和 NF-B 通路相关蛋白的表达。
JDHY 和 BAY 可显著降低 ALF 大鼠血清中 AST、ALT、ALP 和 TBiL 的表达(<0.01),增加 PTA 和 PLT(<0.01),减轻肝组织损伤。此外,JDHY 和 BAY 可减少 ALF 大鼠肝细胞凋亡,促进肝细胞增殖;同时,降低 CD4+细胞和 F4/80 细胞比例,增加 CD8+细胞比例(<0.01)。进一步研究发现,JDHY 和 BAY 可降低 IFN-、IL-6、IL-1 和 TNF-的水平,同时增加 IL-10 和 IL-13 的水平(<0.01)。此外,血清中 sCD163 和肝组织中 CD163 的表达增加(<0.01)。Western blot 结果证实,JDHY 可抑制 ALF 大鼠组织中 NF-B、IK 和 IKK 的磷酸化表达。
JDHY 通过 NF-B 信号通路上调 CD163/sCD163 的水平,从而调节免疫应答,抑制炎症反应,最终改善大鼠的 ALF。