Comparative study on the regulation of AQP3 and AQ4 expression in rats with chronic atrophic gastritis by volatile oil before and after the combination of Atractylodes macrocephala and Panax ginseng
Compatibility is one of the main characteristics of clinical administration of traditional Chinese medicine. By reasonable compatibility and appropriate decoction, the material basis of traditional Chinese medicine decoction pieces will undergo a certain degree of change, thereby achieving the goal of increasing efficiency and reducing toxicity. The specific mechanism can be scientifically explained through the changes in the material basis and biological activity of the effective parts or groups of effective parts before and after compatibility with classical drugs.

Traditional Chinese medicine believes that the basic pathogenesis of chronic atrophic gastritis (CAG) is spleen and stomach qi deficiency. The common treatment in clinical practice is to use drugs that invigorate the spleen and qi. The efficacy of a single drug is often not ideal, and it is often synergistically enhanced by the combination of complementary drugs. Ginseng and Atractylodes macrocephala are the main medicinal pairs used in clinical treatment of spleen and stomach deficiency syndrome. Atractylodes macrocephala strengthens the spleen and qi, dries dampness and promotes diuresis, and is combined with ginseng, which supplements the spleen and qi, nourishes yin and generates fluids. It is used in a balanced and necessary manner. Research has found that prescriptions mainly composed of ginseng and Atractylodes macrocephala can significantly treat chronic atrophic gastritis, such as Sijunzi Tang and Guizhi Ginseng Tang, both of which are formulated in equal quantities at a 1:1 ratio. The volatile oil of Atractylodes macrocephala is the main active ingredient of Atractylodes macrocephala, which has the effects of promoting gastrointestinal motility, anti-inflammatory, anti-tumor, etc. However, it is currently unknown whether it can serve as a material basis for treating chronic atrophic gastritis.

Aquaporins (AQPs) are related to the regulation of water metabolism in the body. According to reports, reduced expression of AQP3 and AQP4 can lead to water loss in the body and exacerbate the degree of atrophy of gastric mucosal glands in CAG. From this, it can be seen that the abnormal expression of AQP3 and AQP4 may be the molecular biology basis of spleen dysfunction and may also be one of the mechanisms of CAG production. Whether the combination of Atractylodes macrocephala volatile oil and ginseng can achieve the goal of treating chronic atrophic gastritis and enhancing efficacy by upregulating the expression of AQP3 and AQP4 remains to be confirmed through research.

Based on this, this experiment uses gas chromatography-mass spectrometry, electron microscopy, and immunohistochemistry techniques to compare the material basis and biological activity (microstructure, ultrastructure, and AQP3, AQP4 expression) of volatile oil components before and after the combination of Atractylodes macrocephala and Panax ginseng, revealing the scientific connotation of rational combination of traditional Chinese medicine.

 

CAG is a chronic digestive system disease caused by multiple factors, with a high risk of developing into gastric cancer. In the global cancer statistics, the incidence rate and related mortality of gastric cancer rank among the top five. Therefore, the diagnosis and treatment of CAG is one of the important means to prevent the occurrence and development of gastric cancer. Traditional Chinese Medicine believes that spleen and stomach qi deficiency is a common pathogenesis of CAG, and clinical treatment often involves a combination of drugs that strengthen the spleen and invigorate qi. Classic medicines have shown significant clinical efficacy in treating chronic obstructive pulmonary disease (CAG) with the combination of Atractylodes macrocephala and Panax ginseng, nourishing the spleen and qi. The volatile oil of Atractylodes macrocephala is the main active ingredient of ginseng medicine. Therefore, this study compared the material basis and biological activity changes of the volatile oil components before and after Atractylodes macrocephala combined with ginseng, and explored the synergistic mechanism of the two drugs in the treatment of CAG.

In terms of the amount of volatile oil extracted, the amount of volatile oil extracted after compatibility was only 1.44mL higher than that before compatibility. This may be due to the increased content of volatile oil in Atractylodes macrocephala caused by compatibility, or the result of the addition of a small amount of ginseng volatile oil or the production of new components. To investigate the reasons for the changes in its material basis, GC-MS analysis was performed on the volatile oils before and after compatibility. Similar to previous studies, ginseng contains very little volatile oil, making it difficult to extract and determine independently. Therefore, this experiment measured and compared the volatile oil of Atractylodes macrocephala and the volatile oil after compatibility as the research objects. The test results showed that the volatile oil components of Atractylodes macrocephala mainly include Atractylodes macrocephala, Eucalyptol 3,7 (11) – diene, Heptane, Docosane, etc. Among them, Atractylodes macrocephala has the highest content, up to 51.67%. After compatibility with ginseng, the relative content of Atractylodes macrocephala decreased to 45.94%, and a small amount of ginseng volatile oil components (such as ginsenoside 1.01%, β – ginsenoside 0.61%, α – coumarin 0.76%, allophyllene 0.59%, and (E) – β – farnesene 0.39%) were added, indicating that the composition of active ingredients in Atractylodes macrocephala after compatibility may vary due to changes in content or the addition of new components. During the compatibility process, the addition of new ingredients and the physical or chemical reactions that occur due to external factors such as boiling temperature, moisture, and oxygen content may be important reasons for changing the pharmacological substance basis to a certain extent. At present, it is widely believed that the combination of multiple components in traditional Chinese medicine can achieve synergistic effects by intervening and improving the pathogenesis through multiple targets and pathways. Previous studies have found that the combination of ginseng and atractylodes can enhance the therapeutic effect of CAG by increasing the content of saponins and the generation of new components. This indicates that changes in the composition of ginseng and atractylodes saponins contribute to the enhancement of drug efficacy, confirming the view that the combination can enhance the synergistic effect of multiple components in traditional Chinese medicine. In addition, volatile oil components are also important active ingredients in ginseng medicine. There are studies showing that the volatile oil of Atractylodes macrocephala combined with ginseng in a 1:1 and 1:2 ratio has stronger antibacterial activity than the volatile oil of Atractylodes macrocephala. It is speculated that the combination of Atractylodes macrocephala and Panax ginseng may alter the activity of the volatile oil in treating CAG due to changes in its chemical composition. But animal experiments are still needed to confirm.

The pathological research results of animal experiments showed that compared with the model group, the number of gastric mucosal gland deformations decreased and the degree of atrophy was significantly reduced in each drug group. The shape and size of gastric pits tended to be regular, the area of inflammatory cell infiltration was significantly reduced, and the pathological score of gastric tissue was also significantly reduced (P<0.05). Moreover, the pathological score of the volatile oil group after compatibility showed a downward trend compared to the Atractylodes macrocephala volatile oil group. The above results indicate that the volatile oil of Atractylodes macrocephala and Panax ginseng showed a good improvement effect on gastric mucosa before and after compatibility, and the therapeutic effect of Panax ginseng after compatibility showed a certain trend of improvement compared to before compatibility. Regarding the reasons, previous studies have shown that atrophy of gastric mucosal glands is the main pathological feature of CAG. This is due to the infiltration and transmembrane migration of inflammatory cells (mainly monocytes and neutrophils) into the gastric mucosa, which enhances cell permeability and leads to water loss. AQPs are closely related to the regulation of water balance in the body, with AQP3 and AQP4 being highly expressed in normal gastrointestinal tissues and lowly expressed in the gastric mucosa of patients with chronic atrophic gastritis, which may be due to water loss in the gastric mucosa. The immunohistochemical results of this experiment showed that compared with the model group, the Atractylodes macrocephala volatile oil group had a trend of enhancing the expression of AQP3 in gastric mucosa and significantly increasing the expression of AQP4 (P<0.05); The expression of AQP3 and AQP4 in gastric mucosa was significantly increased in the combination of volatile oil group (P<0.05). From this, it can be seen that the combination of volatile oils can enhance the expression of AQP3 and AQP4, prevent the reduction of water from interstitial spaces to cells, improve the imbalance of water metabolism in the body, and alleviate pathological manifestations such as gastric mucosal cell atrophy in CAG. This indicates that the dysregulation of AQP3 and AQP4 expression can serve as one of the pathogenesis mechanisms of CAG and may also become a target for the prevention and treatment of CAG. Based on the research results of material basis, the combination of Atractylodes macrocephala and Panax ginseng may enhance the spleen tonifying and drying effects by affecting the volatile oil material basis and water metabolism, thus achieving the goal of enhancing the therapeutic effect of chronic atrophic gastritis. This result provides a scientific basis for elucidating the scientific connotation of rational compatibility of traditional Chinese medicine.

In summary, the volatile oil of Atractylodes macrocephala can alleviate the pathological conditions of gastric tissue in rats with chronic atrophic gastritis, and its effect is enhanced when combined with ginseng. The mechanism may be related to the decrease in the proportion of Atractylodes macrocephala, the small addition of ginseng volatile oil, and the increased expression of aquaporins AQP3 and AQP4. The research results clarify the scientific connotation of the compatibility and efficacy enhancement of traditional Chinese medicine from the perspectives of material basis and pharmacological mechanism, providing a basis for rational clinical application and a reference for similar related traditional Chinese medicine compatibility research.