目的 探讨晕清降压方对痰瘀互结型肥胖性高血压模型大鼠降压减重的影响及可能作用机制.方法 60只SD雄性大鼠随机选取8只为正常对照组,其余大鼠采用小剂量链脲佐菌素+高盐高脂饲料喂养+慢性束缚应激构建痰瘀互结型肥胖性高血压模型,按随机数字表法将造模成功的40只大鼠分为模型组、替米沙坦组、晕清降压方低、中、高剂量组,每组8只.晕清降压方低、中、高剂量组分别予晕清降压方10、20、40 g/(kg·d)灌胃;替米沙坦组予替米沙坦片3.5 mg/(kg·d)灌胃;正常对照组与模型组予蒸馏水10 mL/(kg·d)灌胃,每日1次,连续给药4周.测定各组血压、体质量;酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测血清瘦素(leptin,Lep)、血清血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)水平.蛋白免疫印迹(Western Blot)法检测下丘脑组织磷酸化 Janus 激酶 2(phosphorylated Janus kinase 2,p-JAK2)/Janus 激酶 2(Janus kinase 2,JAK2)、磷酸化细胞信号传导与转录活化因子 3(phosphorylated signal transducers and activators of transcription factor 3,p-STAT3)/细胞信号传导与转录活化因子 3(signal transducers and activators of transcription factor 3,STAT3)、细胞因子信号传导抑制蛋白 3(suppressor of cytokine signaling 3,SOCS3)蛋白表达;实时PCR法检测下丘脑组织血管紧张素Ⅱ受体(angiotensin type 1 receptor,AT1R)、瘦素受体(leptin receptor,LepR)mRNA表达水平.免疫组化检测大鼠下丘脑LepR、AT1R蛋白表达水平.结果 (1)与正常对照组比较,模型组体质量、血压显著升高(P＜0.05);血清Lep、AngⅡ 含量显著升高(P＜0.05);Western Blot 显示 p-JAK2/JAK2、p-STAT3/STAT3 蛋白表达显著下降(P＜0.05),SOCS3蛋白表达显著升高(P＜0.05);实时PCR与免疫组化均显示LepR的表达下降(P＜0.05),AT1R表达升高(P＜0.05).(2)与模型组比较,替米沙坦组体质量、血压均降低(P＜0.05),p-JAK2/JAK2、p-STAT3/STAT3、SOCS3 无统计学差异(P＞0.05),实时 PCR 与免疫组化显示LepR表达无统计学差异(P＞0.05),AT1R表达下降(P＜0.05);晕清降压方低、中、高剂量组体质量、血压均降低(P＜0.05),Lep、AngⅡ 血清含量水平降低(P＜0.05);p-JAK2/JAK2、p-STAT3/STAT3蛋白表达升高,SOCS3蛋白表达下降(P＜0.05);实时PCR与免疫组化显示LepR的表达升高(P＜0.05),AT1R表达下降(P＜0.05),且改善程度与中药浓度呈正相关.(3)与替米沙坦组比较,晕清降压方低、中、高剂量组体质量降低(P＜0.05),晕清降压方高剂量组降压效果无明显差异(P＞0.05);晕清降压方低剂量组Lep升高(P＜0.05),晕清降压方高剂量组Lep降低(P＜0.05),晕清降压方中剂量组Lep无统计学差异(P＞0.05);晕清降压方低、中剂量组AngⅡ升高(P＜0.05),晕清降压方高剂量组AngⅡ无统计学差异(P＞0.05).Western Blot显示晕清降压方低、中、高剂量组p-JAK2/JAK2、p-STAT3/STAT3蛋白表达显著升高(P＜0.05),SOCS3蛋白表达降低(P＜0.05),实时PCR与免疫组化显示晕清降压方低、中、高剂量组LepR均升高(P＜0.05),而晕清降压方高剂量组AT1R无显著差异(P＞0.05).结论 晕清降压方可减轻痰瘀互结型肥胖性高血压模型大鼠体质量、降低血压,其机制可能通过调节JAK2/STAT3信号通路,与降低血管紧张素Ⅱ水平、改善下丘脑瘦素抵抗有关.
Study on the mechanism of Yunqing Jiangya Decoction mediated JAK2/STAT3 pathway improving leptin resistance in obese hypertension
Objective This study aimed to investigate the effects of Yunqing Jiangya Decoction on blood pressure reduction and weight loss in hypertensive obese rats with the phlegm-blood stasis syndrome and explore potential underlying mechanisms.Methods Sixty male Sprague-Dawley(SD)rats were randomly divided into six groups,with eight rats in the control group and the remaining rats subjected to a combination of low-dose STZ,high-salt high-fat diet,and chronic restraint stress to establish a hypertensive obese rat model with phlegm-blood stasis syndrome.The 40 model rats were further randomized into the model group,telmisartan group,and low,medium,and high-dose Yunqing Jiangya Decoction groups,8 rats in each group.Rats in the low,medium,and high-dose Yunqing Jiangya Decoction groups received oral gavage of 10,20,and 40 g/(kg·d)of Yunqing Jiangya Decoction,respectively,while the telmisartan group received telmisartan at a dose of 3.5 mg/(kg·d).The control group and the model group received distilled water 10 ml/(kg·d)by oral gavage.Treatment was administered once daily for four weeks,and blood pressure and body weight were measured.The levels of leptin(Lep)and angiotensin Ⅱ(AngⅡ)in serum were assessed using ELISA.Western blot was used to evaluate the expression of phosphorylated Janus kinase 2/Janus kinase 2(p-JAK2/JAK2),phosphorylated signal transducer and activator of transcription 3/signal transducer and activator of transcription 3(p-STAT3/STAT3),and suppressor of cytokine signaling 3(SOCS3)in the hypothalamic tissues.Real-time PCR was employed to determine the mRNA expression levels of angiotensin Ⅱ receptor(AT1R)and leptin receptor(LepR)in the hypothalamic tissues.Immunohistochemistry was utilized to assess the protein expression levels of LepR and AT1 R in the hypothalamus.Results(1)Compared with the normal control group,the body weight and blood pressure of the model group were significantly increased(P＜0.05).The levels of Lep and AngⅡ in serum increased significantly(P＜0.05).Western Blot showed that the expression of p-JAK2/JAK2,p-STAT3/STAT3 were decreased significantly(P＜0.05),while the expression of SOCS3 were increased significantly(P＜0.05).Real-time PCR and immunohistochemistry showed that the expression of LepR were decreased(P＜0.05)and the expression of AT1 R were increased(P＜0.05).(2)Compared with the model group,the body weight and blood pressure in the telmisartan group were decreased(P＜0.05),and there was no statistical difference in p-JAK2/JAK2,p-STAT3/STAT3 and SOCS3(P＞0.05).Real-time PCR and immunohistochemistry showed no statistical difference in the expression of LepR(P＞0.05),and the expression of AT1R were decreased(P＜0.05).The body weight and blood pressure of low,middle and high doses of Yunqing Jiangya Decoction were decreased(P＜0.05),and the serum levels of Lep and AngⅡ were decreased(P＜0.05).The expression of p-JAK2/JAK2 and p-STAT3/STAT3 protein were increased,while the expression of SOCS3 protein were decreased(P＜0.05).Real-time PCR and immunohistochemistry showed that the expression of LepR increased(P＜0.05)and the expression of AT1 R were decreased(P＜0.05),and the degree of improvement was positively correlated with the concentration of traditional Chinese medicine.(3)Compared with the telmisartan group,the weight of low,middle and high doses of Yunqing Jiangya Decoction were decreased(P＜0.05),but there was no significant difference in antihypertensive effect in high dose of Yunqing Jiangya Decoction(P＞0.05).Lep in the low-dose group of Yunqing Jiangya Decoction were increased(P＜0.05),while it was decreased in the high-dose group of Yunqing Jiangya Decoction(P＜0.05).There was no statistical difference in middle-dose group of Yunqing Jiangya Decoction(P＞0.05).AngⅡ in the low-dose and middle-dose group of Yunqing Jiangya Decoction were increased,but there was no statistical difference in the high-dose group of Yunqing Jiangya Decoction(P＞0.05).Western Blot showed that the expressions of p-JAK2/JAK2,p-STAT3/STAT3 protein in the low,middle and high groups of Yunqing Jiangya Decoction were increased significantly(P＜0.05),while the expression of SOCS3 protein was decreased(P＜0.05).Real-time PCR and immunohistochemistry showed that the LepR in the low,middle and high groups of Yunqing Jiangya Decoction were increased(P＜0.05).Conclusion Yunqing Jiangya Decoction can reduce body weight and blood pressure in hypertensive obese rats with the phlegm-blood stasis syndrome and its mechanism may be related to reducing angiotensin Ⅱ level and improving hypothalamic leptin resistance by regulating JAK2/STAT3 signaling pathway.
obesity hypertensionYunqing Jiangya Decoctionleptin resistanceJAK2/STAT3 signaling pathwaystagnation of phlegm and blood stasissuppressor of cytokine signaling 3
万方数据
维普
