基于斑马鱼模型研究藏药十味蒂达胶囊对代谢功能障碍相关脂肪性肝炎的干预作用及可能机制

白马; 赵鑫杭; 娘毛先; 降拥四郎; 张艺

doi:10.13748/j.cnki.issn1007-7693.20261321

基于斑马鱼模型研究藏药十味蒂达胶囊对代谢功能障碍相关脂肪性肝炎的干预作用及可能机制

Interventional Effects and Potential Mechanisms of Tibetan Medicine Shiwei Dida Capsules on Metabolic Dysfunction-Associated Steatohepatitis Based on Zebrafish Model

摘要

摘要:
目的基于斑马鱼模型评价藏药十味蒂达胶囊对蛋黄粉诱导代谢功能障碍相关脂肪性肝炎(metabolic dysfunction-associated steatohepatitis，MASH)的干预作用并初步探讨其可能机制。
方法采用受精后5天(5 days post-fertilization，5 dpf)的野生型AB品系斑马鱼幼鱼进行最大耐受浓度测定。将斑马鱼随机分为空白组，模型组，阳性对照组及十味蒂达胶囊低、中、高剂量组(37.5、75和150 μg·mL⁻¹)，在蛋黄粉诱导的同时进行药物干预。给药后，采用整体油红O染色评价肝区脂质沉积情况，HE染色观察肝组织病理变化，检测ALT、AST、TC、TG、HDL-C、LDL-C水平，并采用qRT-PCR法检测PPARG、Fasn、LPL、TNF-α和IL-1β mRNA表达。
结果十味蒂达胶囊在150 μg·mL⁻¹及以下浓度对斑马鱼幼鱼未见明显毒性。与空白组比较，模型组斑马鱼肝区油红O染色积分光密度(integrated optical density，IOD)值、ALT、AST、TC、TG、LDL-C水平及PPARG、Fasn、TNF-α、IL-1β mRNA相对表达量均显著升高，HDL-C水平及LPL mRNA相对表达量均显著降低(P<0.05或P<0.001)，病理可见肝细胞肿胀、组织结构紊乱及脂滴样空泡形成。与模型组比较，十味蒂达胶囊各剂量组油红O染色IOD值、ALT、AST、TC及LDL-C水平均降低，HDL-C水平升高；十味蒂达胶囊中、高剂量组TG水平降低(P<0.05或P<0.01)，低剂量组TG水平差异无统计学意义。十味蒂达胶囊各剂量组均可下调IL-1β mRNA表达(P<0.001)，上调LPL mRNA表达(P<0.05或P<0.001)；十味蒂达胶囊中、高剂量组可下调PPARG和TNF-α mRNA表达(P<0.05、P<0.01或P<0.001)，仅高剂量组可下调Fasn mRNA表达(P<0.05)。
结论十味蒂达胶囊对蛋黄粉诱导的斑马鱼MASH具有明确保护作用，其作用可能与改善脂质代谢紊乱及抑制炎症反应相关。

Abstract:
OBJECTIVE To evaluate the intervention effects of Tibetan medicine Shiwei Dida capsules on egg yolk powder-induced metabolic dysfunction-associated steatohepatitis(MASH) based on zebrafish model, and to preliminarily explore its possible mechanism.
METHODS Wild-type AB strain zebrafish larvae at 5 days post-fertilization(5 dpf) were used for the determination of maximum tolerated concentration. Zebrafish were randomly divided into blank group, model group, positive control group, and low-, medium-, and high-dose Shiwei Dida capsule groups(37.5, 75, 150 μg·mL⁻¹). Drug intervention was carried out simultaneously with egg yolk powder induction. After treatment, whole-mount oil red O staining was used to evaluate lipid deposition in the liver region, and HE staining was applied to observe histopathological changes of liver tissue. The levels of ALT, AST, TC, TG, HDL-C, and LDL-C were measured. The mRNA expression levels of PPARG, Fasn, LPL, TNF-α, and IL-1β were determined by qRT-PCR.
RESULTS Shiwei Dida capsules showed no apparent toxicity to zebrafish larvae at concentrations of 150 μg·mL⁻¹ and below. Compared with the blank group, the integrated optical density(IOD) values of oil red O staining in the liver region, the levels of ALT, AST, TC, TG, LDL-C, as well as the relative mRNA expression levels of PPARG, Fasn, TNF-α, and IL-1β were significantly increased in the model group, while the HDL-C level and relative LPL mRNA expression level were significantly decreased(P<0.05 or P<0.001). Hepatocellular swelling, tissue structural disorder and lipid droplet-like vacuolation could be observed pathologically. Compared with the model group, the IOD values of oil red O staining, levels of ALT, AST, TC and LDL-C were decreased and HDL-C level was increased in each dose group of Shiwei Dida capsules. The TG levels were reduced in medium- and high-dose groups of Shiwei Dida capsules(P<0.05 or P<0.01), while there was no statistically significant difference in TG level in the low-dose group. The expression of IL-1β mRNA was downregulated(P<0.001) and the expression of LPL mRNA was upregulated(P<0.05 or P<0.001) in all dose groups of Shiwei Dida capsules. The medium- and high-dose groups of Shiwei Dida capsules could downregulate the mRNA expression of PPARG and TNF-α(P<0.05, P<0.01 or P<0.001), and only the high-dose group could downregulate Fasn mRNA expression(P<0.05).
CONCLUSION Shiwei Dida capsules exert an explicit protective effect against egg yolk powder-induced MASH in zebrafish, and its effect may be related to improving lipid metabolic disorder and inhibiting inflammatory response.

HTML全文

参考文献(37)

施引文献

资源附件(0)