A Self Control Before-after Trial Study Of Shu Gan Yun Pi Decoction On The Treatment Of Obesity-associated Hypertension And Mechanisms Of Astragaloside ?

Objective: The increased occurrence of obesity resultes in continually elevated prevalence of hypertension;however,an effective and targeted treatment drugs have not been found.According to our clinical practice,we thought the pathogenesis of obesity-associated hypertension could be attribute to “the spleen fails to healtily move,and the liver fails to normally dredge and excrete” according to the theory of Traditional Chinese medicine,and the main treatment method is Jian Pi Shu Gan.In this study,we will observe effects of Shu Gan Yun Pi decoction on obesity-associated hypertensive patients.Previous studies have shown that low-grade pro-inflammatory state mediated by the activation of IKK?/NF-kB pathway and leptin resistance are important underlying mechanisms that contribute to obesity-associated hypertension.Activating the cholinergic anti-inflammatory pathway by acting on ?7nAchR is an effective way that inhibiting inflammatory reaction.Astragaloside ?(As ?)is one of the major bioactive compounds extracted from Astragali Radix,studies have shown that peripheral administration of As ? exerted an anti-inflammatory effect through strongly inhibiting the production of pro-inflammatory cytokines and activation of IKK?/ NF-kB pathway in central neural system.Accordingly,we tested the hypothesis that activating ?7nAchR could prevent the occurrence of obesity-associated hypertension by attenuating inflammatory reaction and leptin resistance,and As ? could prevent obesity-associated hypertension by inhibiting pro-inflammatory reaction and leptin resistance;moreover,the effects of As ? might be partly attributed the up-regulated expression of ?7nAchR.Methods:(1)A prospective small sample and self-control before-after trial was conducted to evaluate the effects of Shu Gan Yun Pi decoction on the blood pressure,glucose and lipid metabolism and pro-inflammatory cytokines in obesity-associated hypertensive patients.(2)High-fat diet(HFD)for 16 weeks to induce obese rats and then randomly assigned these rats to three groups: HFD control group(HF con group),?7nAchR agonist group(PNU group),and the ?7nAchR agonist and inhibitor group(PNU+?-BGT group).Rats in the PNU group were intraperitoneally injected with PNU-282987(3mg/Kg/d)for 6 weeks,and rats in the ?-BGT +PNU group were co-administrated with PNU-282987(3mg/Kg/d)and ?-BGT(1?g/Kg/d)intraperitoneally for 6 weeks.Body weight,blood levels of glucose and lipid and blood pressure were measured.NE levels in the plasma and renal cortex were evaluated to reflect sympathetic activity.Protein or mRNA levels of ?7nAChR,IKK? and NF-kB as well as proinflammatory cytokines(IL-1? and TNF-?)in hypothalamus and adipose tissue were detected to reflect the inflammatory statement.Additionally,we tested the expressions of LepRb mRNA,p-STAT3,p-PI3 K,SOCS3 mRNA,and PTP1 B mRNA by Western blot or qRT-PCR to evaluate the hypothalamic leptin sensitivity.(3)HFD induced obese rats were randomly assigned to three groups: the HF con group,As ? group and As ?+?-BGT group.As ?(20 mg/Kg/d)was continually administrated to rats for 6 weeks via daily oral gavage;meanwhile,rats in the As ?+?-BGT group were also intraperitoneal injected with ?-BGT(1?g/Kg/d).The detection indicators were the same as described in the part(2).Results: After treatment with Shu Gan Yun Pi decoction,blood pressure and heart rate were decreased;meanwhile,blood levels of glucose,lipid,hs-CRP and IL-6 decreased when compared with before-treatment.Furthermore,symptoms of Gan Yu Pi Xu syndrome such as dizziness,chest tightness,limbs drowsiness,upset irritability,bloating,body fatigue and loose stools were improved.Also,according to the antihypertensive efficacy evaluation criteria,we divided the patients into valid group and invalid group,the results showed that blood levels of hs-CRP and IL-6 of patients in valid group were improved compared with patients in invalid group.(2)In our animal studies,we found that rats simply administrated with PNU-282987 had lower body weight,adipose tissue weight,size of adipocytes and improved metabolic disorders when compared with rats in the HF con group.Rats in the PNU group had decreased blood pressure and lower levels of NE levels in blood and renal cortex tissue compared with HF con group.In the hypothalamus and adipose tissue,single administration of PNU-282987 increased expression of ?7nAChR while decreased expressions of IKK?,NF-kB and pro-inflammatory cytokines(IL-1? and TNF-?);moreover,hypothalamic expressions of LepRb mRNA and p-STAT3 increased,while p-PI3K?SOCS3 mRNA and PTP1 BmRNA decreased.These results indicated that treatment with PNU-282987 could inhibit inflammatory reaction and improve leptin sensitivity.In contrast,when co-administrated with ?-BGT,all the effects of PNU-282987 decreased,suggesting that activation of ?7nAChR is a crucial underlying mechanism.(3)We found that As ? prevented body weight gain and adipose accumulation,and also improved metabolic disorders in HFD rats.Furthermore,As ? decreased blood pressure and heart rate,as well as NE levels in blood and renal tissue.Meanwhile,treatment with As ? could up-regulate ?7nAchR and suppress IKK?/NF-kB signaling and pro-inflammatory cytokines in the hypothalamus and adipose tissue.In the hypothalamus,As ? alleviated leptin resistance as evidenced by the increased p-STAT3,LepRb mRNA and POMC mRNA,and decreased p-PI3 K,SOCS3 mRNA,and PTP1 B mRNA.Accordingly,we concluded that the effects of As ? on inflammatory reaction and leptin sensitivity were partly related the ?7nAChR,since co-administration of ?7nAChR selective antagonist ?-BGT could weaken the improved effects of As ?.Conclusion:(1)According to our results,we concluded that for obesity-associated hypertensive patients who have 1 and 2 class blood pressure,single treatment with Shu Gan Yun Pi decoction could suppress hypertension,improve metabolism disorders and inflammatory reaction.The underlying mechanism might be related to the suppressed sympathetic activity and reduced inflammatory reaction after treatment.(2)In high-fat diet induced obese rats,activating ?7nAChR could inhibit IKK?/NF-kB activation in the hypothalamic and adipose tissue,which decrease sympathetic over-excitation by preventing the development of inflammatory reaction and leptin resistance.(3)As ? could efficiently prevent obesity-associated hypertension through inhibiting inflammatory reaction and improving leptin resistance;furthermore,these effects of As ? was partly related to the increased ?7nAchR expression.