健脾益氣方對肝癌模型大鼠的治療作用及機制研究

王超 岳紫晨 音金萍 蔣筱 卓少元

中圖分類號 R285.5 文獻標志碼 A 文章編號 1001-0408（2021）19-2342-05

DOI 10.6039/j.issn.1001-0408.2021.19.06

摘 要 目的：探討健脾益氣方對二乙基亞硝胺（DEN）誘導的肝癌模型大鼠的治療作用及機制。方法：將80只雄性SD大鼠分為正常組、模型組、NOD樣受體家族3（NLRP3）抑制劑組（MCC950，4.5 mg/kg）、胱天蛋白酶1（caspase-1）抑制劑組（VX-765，4.5 mg/kg）和健脾益氣方低、中、高劑量組（5.25、10.5、21 g/kg），除模型組20只大鼠外（其中10只用于判斷是否造模成功），其余每組10只。除正常組大鼠腹腔注射生理鹽水外，其余各組大鼠腹腔注射DEN（70 mg/kg）以復制肝癌模型。造模成功后，正常組和模型組大鼠灌胃生理鹽水，各抑制劑組大鼠腹腔注射相應藥物，健脾益氣方各劑量組大鼠灌胃相應藥物，每日1次，連續4周。末次處置后，觀察大鼠肝組織病理學形態變化，檢測大鼠血清中腫瘤壞死因子α（TNF-α）、白細胞介素1β（IL-1β）的含量，檢測大鼠肝組織中NLRP3以及細胞程序性壞死相關蛋白[銜接子凋亡相關斑點樣蛋白（ASC）、caspase-1前體（pro-caspase-1）、受體相互作用蛋白激酶1（RIP1）、RIP3、混合系激酶區域樣蛋白（MLKL）]的表達水平。結果：與正常組比較，模型組大鼠的肝細胞可見不同程度的脂肪變性、細胞核增大和呈團塊狀，部分可見出血和壞死，并伴有增生灶和結節;其肝組織損傷指數和血清中TNF-α、IL-1β含量以及肝組織中NLRP3、ASC、pro-caspase-1、RIP1、RIP3、MLKL蛋白的表達水平均顯著升高（P<0.05或P<0.01）。與模型組比較，健脾益氣方低、中劑量組大鼠的肝組織仍有大量炎性細胞浸潤，而其高劑量組和各抑制劑組大鼠的炎性細胞浸潤均明顯減少;且健脾益氣方各劑量組和各抑制劑組大鼠的肝組織損傷指數和血清以及肝組織中上述指標水平大部分顯著降低（P<0.05或P<0.01）。結論：健脾益氣方對肝癌模型大鼠具有治療作用，其作用機制可能與下調NLRP3炎癥小體的表達、抑制細胞程序性壞死有關。

關鍵詞 健脾益氣方;肝癌;NOD樣受體家族3炎癥小體;程序性壞死

Study on Therapeutic Effects and Its Mechanism of Jianpi Yiqi Decoction on Liver Cancer Model Rats

WANG Chao1，2，YUE Zichen1，YIN Jinping1，JIANG Xiao1，ZHUO Shaoyuan1（1. School of Basic Medicine， Guangxi University of TCM， Nanning 530200， China; 2. Guangxi Key Laboratory of Integrated Traditional Chinese and Western Medicine Translational Medicine for High Incidence Infectious Diseases， Guangxi University of TCM， Nanning 530200， China）

ABSTRACT ? OBJECTIVE： To explore the therapeutic effects and its mechanism of Jianpi yiqi decoction on diethylnitrosamine （DEN） induced liver cancer model rats. METHODS： Totally 80 male SD rats were divided into normal group， model group， Nod-like receptor family 3 （NLRP3） inhibition group （MCC950，4.5 mg/kg）， caspase-1 inhibitory group （VX-765，4.5 mg/kg）， Jianpi yiqi decoction low-dose， medium-dose and high-dose groups （5.25， 10.5， 21 g/kg）， with 10 rats in each group except for 20 rats in model group （10 of them were only used to judge whether modeling was successful）. Rats in each group were intraperitoneally injected with DEN （70 mg/kg） to induce liver cancer model， except for the rats in normal group which were replaced by normal saline. After modeling， normal group and model group were given normal saline intragastrically; inhibitor groups were given relevant medicine intraperitoneally; Jianpi yiqi decoction groups were given relevant medicine intragastrically， once a day， for consecutive 4 weeks. After last administration， histopathological morphology of liver tissue was observed. The contents of serum inflammatory factors TNF-α and IL-1β were detected. The expression of NLRP3 and programmed cell necrosis associated protein （ASC， pro-caspase-1， RIP1， RIP3 and MLKL） in liver tissue were detected. RESULTS： Compared with the normal group， the hepatocytes of model group showed varying degrees of steatosis， enlarged nuclei， lumpy， bleeding and necrosis， accompanied by proliferative foci and nodules. Liver tissue injury index， serum content of TNF-α and IL-1β as well as the protein expression of NLRP3， ASC， pro-caspase-1， RIP1， RIP3 and MLKL in liver tissue were significantly increased （P<0.05 or P<0.01）. Compared with model group， there were still a large number of inflammatory cell infiltration in the liver tissue of rats in Jianpi yiqi decoction low-dose and medium dose groups， while the inflammatory cell infiltration of rats in high-dose group and inhibitor groups decreased significantly; the liver tissue injury index and above indexes levels in serum and liver tissue were decreased significantly （P<0.05 or P<0.01）. CONCLUSIONS： Jianpi yiqi decoction shows therapeutic effect on liver cancer model rats， the mechanism of which may be associated with down-regulating the expression of NLRP3 inflammasome and inhibiting programmed cell necrosis.