Effects of Dachengqi Decoctions Made from Different Processed Products of Rhubarb on ABP Mice

Chunli TANG, Jiangcun WEI, Xiumei MA, Guangli LU, Dongmei HUANG, Sen LIANG, Ling CHEN, Ying YANG, Bihua NONG*

1. The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530023, China; 2. Guangxi International Zhuang Medicine Hospital, Nanning 530201, China

Abstract [Objectives] This study aims to investigate the effects of Dachengqi decoctions made from different processed products of rhubarb on water intake, defecation amount, urination amount, urination volume, dryness of stool, mental state and activity of ABP mice. [Methods] Total 165 mice were randomly divided into Dachengqi decoction groups (made from different processed products of rhubarb, 6 and 10 g/kg), positive control group, blank control group and model group. The administration dosage was 20 mL/kg. With the metabolic cage integral method, the effects of Dachengqi decoctions made from different processed products of rhubarb on the water intake, defecation amount, urination volume, dryness of stool, mental state and activity of ABP mice were compared. [Results] Dachengqi decoctions all could soften stool and promote rapid defecation of the mice with fecal peritonitis of excess heat stagnation type except that made from carbonized rhubarb. The water intakes of all the Dachengqi decoction groups were higher than those of the positive control group and the model group, except the carbonized rhubarb-made Dachengqi decoction groups. [Conclusions] Dachengqi decoctions all have obvious purgative effect except those made from carbonized rhubarb.

Key words Rhubarb, Dachengqi decoction, Processed product, Fecal peritonitis

1 Introduction

Dachengqi decoction was originally published in Zhang Zhongjing’sTreatiseonFebrileDiseasesandSynopsisofPrescriptionsoftheGoldenChamberin the Han Dynasty. It is composed of rhubarb, Fructus Aurantii Immaturus, Officinal Magnolia Bark and mirabilite, and is the representative prescription of purgation[1-2]. Dachengqi decoction has the effect of purging intense heat, so it is mainly used to treat Yangming fu-viscera excess and excess interior heat[3]. Clinically, it is mainly used for the treatment of diseases such as acute abdomen, intestinal obstruction and gastrointestinal dysfunction. In addition, the prescription also has a good effect in improving the awareness of cerebrovascular disease patients and the rescue of organophosphorus pesticide acute poisoning[4-5]. Research[4]shows Dachengqi decoction has the effects of purging, fighting inflammation, reducing inflammatory cytokines, inhibiting serum endotoxins and regulating the secretion of gastrointestinal hormones, and has obvious protective effects on brain, lung and other organs. Rhubarb is a monarch medicine in Dachengqi decoction, and its safety is determined by the Chinese herbs that make up the prescription. After processing, the properties of Chinese herbs will change, and the toxic and side effects will also be reduced[6].

After processing by different methods, the chemical composition and efficacy of rhubarb will change greatly, and opposite effect may even produce[7-8]. Current reports mostly focus on the changes in the efficacy of rhubarb after its compatibility with other Chinese herbs[9-10]. The changes in the purgative effect of rhubarb after its compatibility with other Chinese herbs and the common relationship reflected in different compatibilities have not been mentioned. In this study, the effects of Dachengqi decoctions made from different processed products of rhubarb on mice with fecal peritonitis were investigated.

2 Materials and methods

2.1Materials

2.1.1Instruments and equipment. Metabolic cage (ZH-B6, Anhui Zhenghua Co., Ltd.); incubator (PYX-190H-B, Guangdong Keli Co., Ltd.); electronic analytical balance[PRACTUM224-KN SOP, Sartorius Scientific Instruments (Beijing) Co., Ltd.]; Centrifuge (41284881, Thermo Fisher, Germany); water bath (HWS12, Shanghai Yiheng Scientific Instrument Co., Ltd.).

2.1.2Medicinal herbs. The used medicinal herbs included rhubarb (Origin: Sichuan; batch No.180307001), Fructus Aurantii Immaturus (Origin: Jiangxi; batch No.180101771), Officinal Magnolia Bark (Origin: Shaanxi; batch No.180507021) and mirabilite (Origin: Qinghai; batch No.180307611). They were purchased from Guangdong Kangmei Pharmaceutical Co., Ltd., and were identified by the chief Chinese pharmacist of the First Affiliated Hospital of Guangxi University of Chinese Medicine. Rhubarb was the dried root and rhizome ofRheumofficinaleBaill. (Polygonaceae); Fructus Aurantii Immaturus was the dried young fruit ofCitrusaurantiumL. (Rutaceae); Officinal Magnolia Bark was the dried trunk bark, root bark and branch bark ofMagnoliaofficinalisRehd. et Wils. (Magnoliaceae). The above three traditional Chinese medicines were crushed and passed through 10-mesh sieve. Mirabilite, present in the form of crystal, is a sulfate-based mineral, and it mainly contains sodium sulfate decahydrate (Na2SO4·10H2O).

2.1.3Animals. The test animals were clean-grade Kunming mice[half male and half female, body weight (20±2) g], which were provided by the Experimental Animal Center of Guangxi Medical University (animal quality certification 0001757, production license SCXK Gui 2014-0002).

2.2Methods

2.2.1Preparation of test decoctions. According to the prescription, accurate amounts of powder of Officinal Magnolia Bark (24 g), Fructus Aurantii Immaturus (12 g), rhubarb (12 g) and mirabilite (9 g) were weighed. The powder of Officinal Magnolia Bark and Fructus Aurantii Immaturus was mixed, soaked in 500 mL of pure water for 30 min, kept slightly boiling for 30 min and filtered through a 500-mesh filter cloth, and the residue obtained was added with 500 mL of pure water, soaked for 30 min, boiled slightly for 30 min and filtered. The filtrates of the two times were merged, added with powder of rhubarb, soaked for 20 min, boiled slightly for 20 min and filtered through 500-mesh filter cloth, and the residue obtained was added with 300 mL of pure water, boiled slightly for 20 min, and filtered. The filtrates of the two times were merged, added with mirabilite while hot, stirred to make the mirabilite completely dissolve, concentrated into a 0.3∶1 liquid (containing 0.3 g/mL of crude drug) and stored in the refrigerator at 2-8℃ for future use.

In the same way as Section2.2.1, a liquid of 0.5∶1 (containing 0.5 g/mL of crude drug) was prepared and stored in the refrigerator at 2-8 ℃ for future use.

Dachengqi decoctions made from different processed products of rhubarb, raw rhubarb (RDCQD), stewed rhubarb (SDCQD), wine-treated rhubarb (WDCQD) and carbonized rhubarb (CDCQD) were prepared.

2.2.2Establishment of ABP mouse model. With reference to literature[11]and combined with the results of pre-experiment, 15 healthy mice were randomly selected as a normal control group, and each of them was injected intraperitoneally with 0.5 mL of sodium chloride. A certain amount (6 g) of mouse feces was collected, added with 100 mL of physiological saline, ground evenly and let stand for 30 min, and the upper suspension was collected and injected intraperitoneally to the mice according to a dosage of 0.5 mL/mouse. After 20 h of the intraperitoneal injection, the mice developed thick secretion in the corners of the eyes, purple lips, less movement, erected hair, crouching, bloating in the abdomen, lack of appetite and extremely dry stools. After dissection, it could be observed that there was a small amount of turbid liquid in the abdominal cavity, most of the intestine swelled, the intestinal tract was congested, and some part of the intestinal tract cramped, indicating successful modeling. For Yangming fu-viscera excess, cold purgation method can be used clinically.

2.2.3Grouping and administration. After 3 d of adaptive feeding, 165 healthy mice weighing (20±2) g, half male and half male, were randomly and evenly divided into the following 11 groups: (a) 6 and 10 g/kg RDCQD groups; (b) 6 and 10 g/kg SDCQD groups; (c) 6 and 10 g/kg WDCQD groups; (d) 6 and 10 g/kg CDCQD groups; (e) blank control group; (f) model group; (g) positive control group (ampicillin, 20 mL/kg body weight, 60 mg/mL). After successful modeling, the mice in different doses of DCQD groups were administered once a day, for two consecutive days. The mice in the positive control group were given with ampicillin after successful modeling according to the dosage of 0.06 g/mL and 20 mL/kg, once a day, for two consecutive days. In the blank control group, after 20 h of intraperitoneal injection of physiological saline (20 mL/kg), administration of distilled water by gavage was performed, once a day, for two consecutive days. After successful modeling, distilled water was intragastrically administered to the mice in the model group, 20 mL/kg, once a day, for two consecutive days. After administration, the water intake, urination volume, defecation amount, dryness of stool, mental state and activity of the mice were observed, and the levels of NO and tumor necrosis factor-α (NTF-α) in the mice were determined.

2.2.4Sample collection and processing. After administration, the water intake, defecation amount, urination volume, dryness of stool, mental state and activity of the mice were observed. Within 24 h after administration, the water intake and defecation amount of the mice in the treatment groups were observed.

2.2.5Statistical processing. The experimental data are expressed as mean ± standard deviation. SPSS 21.0 statistical software was used for data analysis. One-way analysis of variance was used for the comparison of the mean of multiple groups in a completely random design. If the difference was significant, SNK-q test was performed for multiple comparisons among multiple samples. WhenP<0.05, difference is considered significant.

3 Results and analysis

3.1GeneralsituationandmortalityofmicewithfecalperitonitisofexcessheatstagnationtypeFrom the beginning of the modeling to the end of the experiment, the symptoms, signs and mortality of the mice in each group were observed and analyzed. The results are shown in Table 1.

As shown in Table 1, after modeling, the symptom of Yangming fu-viscera excess appeared in the mice of the model group, indicating that this modeling method is simple and convenient, and can be applied to the following pharmacological tests, regardless of its high mortality.

Table 1 General situation and mortality of the mice in each group after modeling (n=15)

3.2EffectonwaterintakeofthemiceThe water intake of the mice was analyzed in June, when the temperature of the animal house was (26±2)℃, and the air humidity was around 50%.

Table 2 shows that there were significant differences in 24-h water intake between the blank control group and the 6 g/kg RDCQD group, 10 g/kg RDCQD group, 6 g/kg SDCQD group, 10 g/kg SDCQD group, 6 g/kg WDCQD group, 10 g/kg WDCQD group, 6 g/kg CDCQD group, 10 g/kg CDCQD group, model group and positive control group (P<0.01).

Table 2 Effects of Dachengqi decoctions made from different processed products of rhubarb on 24-h water intake and 24-h defecation amount of ABP mice n=15)

Compared with that of the model group, the 24-h water intakes of the 6 g/kg RDCQD group, 10 g/kg RDCQD group, 6 g/kg SDCQD group, 10 g/kg SDCQD group, 6 g/kg WDCQD group, 10 g/kg WDCQD group, 6 g/kg CDCQD group, 10 g/kg CDCQD group, blank control group and positive control group were significantly different (P<0.01).

There were significant differences in 24-h water intake between 10 g/kg SDCQD group and RDCQD groups (P<0.05), and between 6 g/kg CDCQD group, 10 g/kg CDCQD group, blank control group, model group, positive control group and RDCQD groups (P<0.01).

3.3EffectondefecationamountofthemiceThe feces in the RDCQD groups, the SDCQD groups, and the WDCQD groups were watery, and their defecation amounts could not be accurately measured.

Compared with that of blank control group, the 24-h defecation amounts of 6 g/kg RDCQD group, 6 g/kg SDCQD group, 6 g/kg WDCQD group, 6 g/kg CDCQD group, 10 g/kg CDCQD group, model group and positive control group were significantly different (P<0.01).

There were significant differences in 24-h defecation amount between 6 g/kg RDCQD group, 6 g/kg SDCQD group, 6 g/kg WDCQD group, 10 g/kg CDCQD group, blank control group and model group (P<0.01).

The 24-h defecation amounts of 6 g/kg CDCQD group, blank control group, model group and positive control group were significantly different from those of RDCQD groups (P<0.01) (Table 2).

4 Discussion

Related research shows that acute infectious diseases and acute abdomen are manifestations of biological activity of endotoxins, or caused by their induced cytokines and other inflammatory mediators. Constipation of excess heat type is a kind Yangming fu-viscera excess, and endotoxin (ET) is one of its causative factors.

The generation and release of tumor necrosis factor is the body’s response to various exogenous or endogenous substances. Both endotoxin and tumor necrosis factor can cause harm to the body. The combination of the two can produce a synergistic effect. Tumor necrosis factor can enter the blood circulation through the role of endocrine, and can also affect the adjacent intestinal epithelial cells in the form of paracrine, thus producing a certain impact on the barrier function of the intestine.

In this study, the effects of Dachengqi decoctions made from different processed products of rhubarb on 24-h water intake and 24-h defecation amount of mice of fecal peritonitis of excess heat stagnation type were investigated. The results show that the Dachengqi decoctions all can soften stools and promote the rapid defecation of fecal peritonitis mice of excess heat stagnation type, except that made from carbonized rhubarb, so that the dry feces in the mice can be quickly discharged and the generation of more heat is avoided.

The water intakes of all the DCQD groups, except CDCQD group, were higher than those of the positive control group and model group. The reason may be that Dachengqi decoction contains Officinal Magnolia Bark and Fructus Aurantii Immaturus, which are all medicines for promoting qi circulation, so that the water intake of the experimental animals was influenced.

Although the Dachengqi decoction made from carbonized rhubarb also contains Officinal Magnolia Bark and Fructus Aurantii Immaturus, carbonized rhubarb is the monarch medicine in the prescription, and it may have no obvious improving effect on the symptoms of excess heat stagnation-typed fecal peritonitis in mice.