葛根素通过调控细胞焦亡改善对野百合碱诱导的肺动脉高压大鼠的影响

Effects of Puerarin on Monocrotaline-Induced Pulmonary Arterial Hypertension in Rats by Regulating Cell Death

目的从细胞焦亡角度研究葛根素对肺动脉高压的作用及机制。方法取50只SD大鼠,随机分为正常对照组、模型组、10 mg/kg葛根素组、20 mg/kg葛根素组、40 mg/kg葛根素组,每组10只大鼠,正常对照组腹腔注射生理盐水,其他组以单次腹腔注射0.06 g/kg(6 mL/kg)的野百合碱构建肺动脉高压模型,构建成功后10 mg/kg葛根素组、20 mg/kg葛根素组、40 mg/kg葛根素组分别以10 mg/kg、20 mg/kg、40 mg/kg的葛根素溶液连续进行腹腔注射28 d进行治疗;通过测压导管检测PASP、mPAP、PAWP及RVIH;通过HE染色检测大鼠血管壁面积比;通过Masson染色检测大鼠肺部胶原沉积情况;采用免疫组织化学、Western blot检测焦亡相关指标Caspase-1、NLRP-3、IL-1β、IL-18的表达情况。结果与正常对照组相比,用野百合碱构建的肺动脉高压模型组大鼠PASP、mPAP、PAWP及RVIH显著增高,相对于模型组来说,10 mg/kg、20 mg/kg、40 mg/kg葛根素组PASP、mPAP、PAWP及RVIH显著降低,并且随着葛根素浓度的增高,下降的程度越明显;与正常对照组相比,模型组大鼠血管壁面积比显著增高,相对于模型组来说,10 mg/kg、20 mg/kg、40 mg/kg葛根素组的血管壁面积比显著降低,并且随着葛根素浓度的增高,下降的程度越明显;与正常对照组相比,模型组大鼠胶原沉积的面积明显增大,与模型组相比,10 mg/kg、20 mg/kg、40 mg/kg葛根素组大鼠胶原沉积的面积相对减少;相对于正常组,模型组大鼠Capase-1、NLRP-3、IL-1β、IL-18表达显著升高,10 mg/kg葛根素组大鼠与模型组对比上述指标差异无统计学意义,而20 mg/kg、40 mg/kg葛根素组与模型组相比,差异有统计学意义。结论葛根素通过降低焦亡程度改善大鼠肺动脉高压的症状,该研究对于肺动脉高压的治疗及研究方向具有重要意义。

Objective To study the effect and mechanism of puerarin on pulmonary arterial hypertension from the perspective of cell pyrolysis.Methods:Fifty SD rats were randomly divided into normal control group,model group,10 mg/kg puerarin group,20 mg/kg puerarin group and 40 mg/kg puerarin group,10 rats in each group.The normal control group was injected with normal saline intraperitoneally,and the other groups were injected with monocrotaline of 0.06 g/kg(6 mL/kg)in a single intraperitoneal injection to construct the pulmonary arterial hypertension model.After the successful construction,the 10 mg/kg puerarin group,20 mg/kg puerarin group and 40 mg/kg puerarin group were injected intraperitoneally with 10 mg/kg,20 mg/kg and 40 mg/kg puerarin solution for 28 days.The levels of pulmonary artery systolic pressure(PASP),mean pulmonary artery pressure(mPAP),pulmonary arteriole wedge pressure(PAWP)and right heart hypertrophy index(RVIH)were detected by pressure measuring catheter.The vascular wall area ratio of rats was detected by HE staining.The collagen deposition in the lungs of rats was detected by Masson staining.The expressions of Caspase-1,NOD-like receptor protein 3(NLRP-3),interleukin-1β(IL-1β)and interleukin-18(IL-18)were detected by immunohistochemistry and Western blot.Results Compared with those of the normal control group,the levels of PASP,mPAP,PAWP and RVIH of the pulmonary hypertension model group constructed with monocrotaline were significantly increased.Compared with those of the model group,the levels of PASP,mPAP,PAWP and RVIH of 10 mg/kg,20 mg/kg,40 mg/kg puerarin groups were significantly reduced,and with the increase of puerarin concentration,the degree of decrease was more obvious.Compared with that of the normal control group,the ratio of the vascular wall area of rats in the model group was significantly higher than that of the model group.In other words,the blood vessel wall area ratio of the 10 mg/kg,20 mg/kg and 40 mg/kg puerarin groups were significantly decreased,and as the puerarin concentration increased,the degree of decrease was more obvious.Compared with that of the normal control group,the area of collagen deposition of the model group rats was significantly increased.Compared with that of the model group,the collagen deposition area of rats in the 10 mg/kg,20 mg/kg and 40 mg/kg puerarin groups was relatively reduced.Compared with those of the normal group,the expressions of Caspase-1,NLRP-3,IL-1βand IL-18 were significantly increased in the model group.There was no significant difference in the above indicators between the 10 mg/kg puerarin group and the model group,while those of the 20 mg/kg and 40 mg/kg puerarin groups were similar to those of the model group.And the difference was significant.Conclusion Puerarin can improve the symptoms of pulmonary arterial hypertension in rats by reducing the degree of pyrolysis.This study is of great significance for the treatment and research direction of pulmonary hypertension.