Objective

2

To explore the potential action targets of typical ascending and floating Chinese medicinals and descending and sinking medicinals.

Methods

2

Firstly， the targets of ascending and floating Chinese medicinals including Mahuang （

Herba Ephedrae

）， Chaihu （

Radix Bupleuri

）， Shengma （

Rhizoma Cimicifuga

）， Gegen （

Radix Puerariae Lobatae

）， Jiegeng （

Radix Platycodonis

）， Bohe （

Herba Menthae

）， and descending and sinking Chinese medicinals including Tinglizi （

Semen Lepidii

）， Xuanfuhua （

Flos Inulae

）， Daodou （

Semen Canavaliae

）， Chuanniuxi （

Radix Cyathulae

）， Fuling （

Poria

）， Kuxinren （

Semen Armeniacae Amarum

）， and Dingxiang （

Flos Caryophylli

） were searched and selected through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform. Disease targets were searched in GeneCards database with the keywords “wind-cold exterior syndrome” and “wind-water edema” （the disease is located at the exterior）， "phlegm-rheum retention” and “pulmonary edema” （the disease is located at the interior）， “down-bearing stomachache” and “heart failure” （sinking tendency of disease）， and “vomitting” and “liver-yang hyperactivity” （upward reversal of disease tendency）. The intersection genes of ascending and floating Chinese medicinals and the diseases location at the exterior and sinking tendency of disease may be the potential targets for the ascending and floating Chinese medicinals； the intersection genes of descending and sinking Chinese medicinals and the disease location at the interior and the upward reversal of disease tendency may be the potential targets for descending and sinking Chinese medicinals. Animal validation experiment： 140 male SD rats were divided into normal control group， Mahuang （4.2 g/kg） group， Chaihu （7 g/kg） group， Shengma （4.2 g/kg） group， Gegen （10.5 g/kg） group， Jiegeng （7 g/kg） group， Bohe （4.2 g/kg） group， Tinglizi （7 g/kg） group， Xuanfuhua （6.3 g/kg） group， Daodou （6.3 g/kg） group， Chuanniuxi （7 g/kg） group， Fuling （10.5 g/kg） group， Kuxingren （7 g/kg） group and Dingxiang （2.1 g/kg） group， with 10 rats in each group. Corresponding Chinese medicinals were given by gavage at the volume of 1 ml/100 g for 14 days. According to the results of network pharmacology， real-time quantitative PCR was used to detect the mRNA levels of insulin-like growth factor 2 （IGF-2）， hexokinase2 （HK-2）， peroxisome proliferator-activated receptor γ （PPAR-γ）， cholinergic receptor muscarinic 3 （CHRM3）， muscarinic cholinergic receptor 1 （CHRM1） and glycogen synthase kinase-3β （GSK-3β）. Western Blot and immunohistochemical methods were used to detect the protein levels of CHRM3， CHRM1 and PPAR-γ.

Results

2

Using network pharmacology， the potential targets of ascending and floating medicinals were IGF-2 and HK-2， and those of descending and sinking medicinals were PPAR-γ， CHRM3， CHRM1 and GSK-3β. The detection of mRNA and proteins showed that compared to normal control group， there were significantly lower CHRM3 levels in Mahuang group， Chaihu group， Shengma group， Gegen group， Jiegeng group， Dingxiang group and Daodou group， while higher CHRM3 level in Kuxingren group （

P

＜0.05 or

P

＜0.01）； Gegen and Jiegeng groups had decreased level of CHRM1， while Bohe， Tinglizi， Xuanfuhua， Chuanniuxi， Fuling， Dingxiang， Kuxingren and Daodou groups had increased level of CHRM1 （

P

＜0.05 or

P

＜0.01）； Mahuang， Shengma， Gegen and Jiegeng groups could significantly decrease the level of PPAR-γ， while Xuanfuhua， Chuanniuxi， Fuling and Kuxingren groups could increase the level of PPAR-γ （

P

＜0.05 or

P

＜0.01）.

Conclusion

2

The potential targets of typical ascending and floating as well as descending and sinking Chinese medicinals are mainly CHRM3， CHRM1 and PPAR-γ.