目的 探討白芍總苷通過調(diào)節(jié)Ras同源基因家族成員A/Rho相關(guān)卷曲螺旋蛋白激酶1（RhoA/ROCK1）信號通路對大鼠肝缺血再灌注損傷的影響。方法 通過阻斷肝動脈和門靜脈1 h建立肝缺血再灌注損傷大鼠模型，設(shè)假手術(shù)組、模型組、白芍總苷（100 mg/kg）組、白芍總苷＋Rhosin（100 mg/kg＋40 mg/kg）組和白芍總苷＋LPA（100 mg/kg＋1 mg/kg）組，每組8只。連續(xù)治療6周后，檢測各組大鼠血清肝功能指標(biāo)[丙氨酸氨基轉(zhuǎn)移酶（ALT）、天冬氨酸氨基轉(zhuǎn)移酶（AST）、總膽紅素（TBiL）]；HE、TUNEL染色法觀察肝組織病理改變和細胞凋亡，并計算肝損傷評分和凋亡指數(shù)（AI）；分光光度法檢測肝組織中氧化應(yīng)激指標(biāo)[超氧化物歧化酶（SOD）、過氧化氫酶（CAT）活性和丙二醛（MDA）含量]，ELISA檢測炎癥因子[白細胞介素（IL）-1β、IL-6、腫瘤壞死因子-α（TNF-α）]含量；通過反轉(zhuǎn)錄聚合酶鏈反應(yīng)（RT-PCR）、蛋白免疫印跡（Western blotting）法檢測肝組織中RhoA、ROCK1、核因子-κB p65（NF-κB p65）、B細胞淋巴瘤-2（Bcl-2）、Bcl-2相關(guān)X蛋白（Bax）、活化型半胱氨酸蛋白酶-3（C-Caspase-3）mRNA和蛋白表達。結(jié)果 與模型組比較，白芍總苷組和白芍總苷＋Rhosin組血清ALT、AST、TBiL水平顯著降低（P＜0.05）；肝組織病理改變和細胞凋亡狀況均明顯改善，肝損傷評分和細胞AI均顯著降低（P＜0.05）；SOD、CAT活性顯著升高，MDA、IL-1β、IL-6、TNF-α含量顯著降低（P＜0.05）；RhoA、ROCK1、NF-κB p65、Bax、C-Caspase-3 mRNA和蛋白表達均顯著降低，Bcl-2 mRNA和蛋白表達均顯著升高（P＜0.05）。與白芍總苷組相比，Rhosin可顯著增強白芍總苷對模型大鼠肝功能、肝組織病變、細胞凋亡、氧化應(yīng)激、炎癥及RhoA/ROCK1信號通路相關(guān)mRNA和蛋白表達的作用；LPA則顯著逆轉(zhuǎn)了上述調(diào)控作用。結(jié)論 白芍總苷可能通過抑制RhoA/ROCK1信號通路，減輕氧化應(yīng)激、炎癥和細胞凋亡，從而對大鼠肝缺血再灌注損傷起到一定的保護作用。

Objective To investigate the effect of total glucosides of paeony on hepatic ischemia-reperfusion injury in rats by regulating RhoA/Rock1 signaling pathway. Methods The rat model with hepatic ischemia-reperfusion injury was established by blocking hepatic artery and portal vein for 1 h. And the sham operation group, model group, total glucosides of paeony (100 mg/kg) group, total glucosides of paeony + TGP (100 mg/kg + 40 mg/kg) group, and total glucosides of paeony + LPA (100 mg/kg + 1 mg/kg) group were set up, with 8 rats in each group. After 6 weeks of continuous treatment, the level of ALT, AST, TBiL in serum were measured, the hepatic tissue pathological changes and cell apoptosis were observed through HE or TUNEL staining, the hepatic injury score and apoptosis index (AI) were calculated. The oxidative stress indexes (SOD, CAT, and the content of MDA) in hepatic tissue were detected by spectrophotometer. The level of inflammatory factors (IL-1β, IL-6, TNF-α) were detected by ELISA method. The mRNA and protein expressions of RhoA, ROCK1, NF-κB p65, Bcl-2, Bax, C-Caspase-3 were in hepatic tissue were detected by reverse transcription-polymerase chain reaction (RT-PCR) or Western blotting. Results Compared with model group, the level of ALT, AST, TBiL in serum of total glucosides of paeony group and total glucosides of paeony + Rhosin group were significantly decreased (P < 0.05). The pathological changes and cell apoptosis of hepatic tissue were significantly improved, the hepatic injury score and AI were significantly decreased (P < 0.05). The activity of SOD, CAT were significantly increased, and the content of MDA, IL-1β, IL-6, TNF-α were significantly decreased (P < 0.05). The mRNA and protein expression of RhoA, ROCK1, NF-κB p65, Bax, C-Caspase-3 were significantly decreased, while the mRNA and protein expression of Bcl-2 were significantly increased (P < 0.05). Compared with total glucosides of paeony group, Rhosin could significantly enhance the effect of total glucosides of paeony on hepatic function, hepatic lesion, apoptosis, oxidative stress, inflammation, the expression of RhoA/ROCK1 signaling pathway-related mRNA and protein in HIR rats, while LPA could reversed the regulatory effect. Conclusion Total glucosides of paeony can alleviate oxidative stress, inflammation, cell apoptosis by inhibiting the RhoA/ROCK1 signaling pathway, thus playing a certain protective role against hepatic ischemia-reperfusion injury in rats.

R285

河北省醫(yī)學(xué)科學(xué)研究課題計劃（20221561）