PPAR-γ依赖的何首乌免疫性特异质肝损伤机制研究

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基于脂多糖(lipopolysaccharide,LPS)复制的免疫性特异质肝损伤模型,考察过氧化物酶体增殖物活化受体-γ(peroxisome proliferator-activated receptorγ,PPAR-γ)对何首乌肝损伤的影响及机制。将70只SpragueDawley(SD)大鼠随机均分为对照组、LPS组(2.8 mg·kg~(-1))、何首乌组(生药2.16 g·kg~(-1))、PPAR-γ激动剂组(0.5mg·kg~(-1))、PPAR-γ激动剂+LPS组(0.5 mg·kg~(-1)、2.8 mg·kg~(-1))、何首乌+LPS组(生药2.16 g·kg~(-1)、2.8 mg·kg~(-1))及何首乌+LPS+PPAR-γ激动剂组(生药2.16 g·kg~(-1)、2.8 mg·kg~(-1)、0.5 mg·kg~(-1))。按组别分别灌胃给予PPAR-γ激动剂,每日1次,连续给药2天,第3天除对照组灌胃等量蒸馏水外,按组别分别灌胃何首乌,3 h后按组别尾静脉注射LPS,7 h后采用戊巴比妥钠将大鼠麻醉,下腔静脉取血并采集肝组织标本,检测血浆丙氨酸转氨酶(alanine transaminase,ALT)和天冬氨酸转氨酶(aspartate aminotransferase,AST),检测血浆肿瘤坏死因子-α(tumor necrosis factor,TNF-α)、白细胞介素-1β(interleukin-1β)、白细胞介素-6(interleukin-6)和干扰素-γ(interferon-γ),肝组织切片检查病理学改变和肝细胞凋亡,免疫组化染色观察肝组织切片PPAR-γ和核因子-κB(nuclear factor-κB,NF-κB)p65的表达。结果显示,肝组织PPAR-γ表达量与何首乌免疫性特异质肝损伤呈负相关,给予PPAR-γ激动剂可显著降低何首乌特异质肝损伤大鼠血浆中ALT和AST水平(均P<0.05),减轻肝组织病理损伤和肝细胞凋亡,显著促进肝组织PPAR-γ的表达并抑制NF-κB p65的表达(均P<0.05),同时显著降低血浆中TNF-α等炎症因子含量(均P<0.05)。研究结果提示,何首乌免疫性特异质肝损伤的发生与PPAR-γ通路异常抑制和相关炎症因子过表达有关,PPAR-γ激动剂可逆转何首乌特异质肝损伤,为阐释何首乌特异质肝损伤机制和寻找配伍减毒药物提供了参考依据。 To investigate the effect of peroxisome proliferator-activated receptorγ (PPAR-γ) on hepatic injury in Polygonum multiflorum Thunb., A liver injury model based on lipopolysaccharide (LPS) replication-specific liver injury. mechanism. Seventy Sprague Dawley (SD) rats were randomly divided into control group, LPS group (2.16 g · kg -1), PPAR-γ agonist (0.5 mg · kg -1), PPARγ agonist + LPS group (0.5 mg · kg -1, 2.8 mg · kg -1), Polygonum multiflorum Thunb. + LPS group (crude drug 2.16 g · kg -1, 2.8 mg · kg -1) and Polygonum multiflorum L. + LPS + PPAR-γ agonist group (crude drug 2.16 g · kg -1, 2.8 mg · kg -1 ), 0.5 mg · kg -1). According to the group, PPAR-γ agonist was given orally by gavage once daily for 2 days. On the 3rd day except for the control group, the same amount of distilled water was orally administered, The rats were anesthetized with sodium pentobarbital 7 hours later, blood was taken from inferior vena cava and samples of liver tissue were collected to detect plasma alanine transaminase (ALT) and aspartate aminotransferase aspartate aminotransferase (AST)) were detected by real-time PCR. The levels of plasma tumor necrosis factor-α, interleukin-1β, interleukin-6 and interferon- The expression of PPAR-γ and NF-κB p65 in hepatic tissue sections were observed by immunohistochemical staining. The results showed that the expression of PPAR-γ in liver tissue was negatively correlated with immune-specific liver damage in Polygonum multiflorum Thunb. The PPAR-γ agonist could significantly decrease the levels of ALT and AST in the plasma of Polygonum multiflorum liver (all P < 0.05), reduce the pathological damage of liver tissue and hepatocyte apoptosis, significantly promote the expression of PPAR-γ in liver tissue and inhibit the expression of NF-κB p65 (all P <0.05), and at the same time significantly decrease the content of inflammatory cytokines such as TNF- (All P <0.05). The results suggest that Polygonum multiflorum autoimmune liver damage is associated with abnormal PPAR-γ pathway and the over-expression of inflammatory cytokines. PPAR-γ agonist can reverse Hepatic allogeneic liver injury, Injury mechanism and find the compatibility of attenuated drugs provides a reference.
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