Korean J Gastroenterol.  2018 Mar;71(3):132-142. 10.4166/kjg.2018.71.3.132.

Combined Extracts of Artemisia and Green Tea, Mitigated Alcoholic Gastritis Via Enhanced Heat-shock Protein 27

• Kim YS
• Jeong M
• Han YM
• Park JM
• Kwon SO
• Hong SP
• Hahm KB

• KMID: 2407670
• DOI: http://doi.org/10.4166/kjg.2018.71.3.132

Abstract

BACKGROUND/AIMS
Several lines of evidence from epidemiologic and laboratory studies have shown that the consumption of Artemisia or green tea extracts (MPGT) is inversely associated with the risk of alcohol-induced damage and other chronic diseases. Supported by previous studies showing that the combined extract of Artemisia and green tea, MPGT, exerted significantly either antioxidative or anti-inflammatory actions against Helicobacter pylori-associated gastric diseases, it was hypothesized that MPGT can offer protection against alcoholic gastritis.
METHODS
Ethanol was administered to induce gastric damage in Wistar rats, which had been pretreated with various doses of MPGT, to measure the rescuing action of a MPGT pretreatment against ethanol-induced gastric damage. In addition, the molecular mechanisms for the preventive effects were examined.
RESULTS
The MPGT pretreatment (100, 300, and 500 mg/kg) alleviated the ethanol-induced gastric damage, which was evidenced by the significant decrease in calcium-dependent phospholipase A2, MAPKs, and NF-κB levels compared to ethanol alone. Furthermore, the MPGT pretreatment preserved 15-prostaglandin dehydrogenase, whereas cyclooxygenase-2 was decreased significantly. All of these biochemical changes led to the significant alleviation of alcohol-associated gastric mucosal damage. Ethanol significantly increased the TUNEL positivity in the stomach, but MPGT decreased the apoptotic index significantly, which was associated with significantly lower pathological scores of ethanol-induced mucosal ulcerations. The significant protective changes observed alcoholic gastritis with MPGT were related to the increased expression of cytoprotective genes, such as heat-shock protein (HSP)27, HSP60, and PDGF.
CONCLUSIONS
The efficient anti-inflammatory, anti-apoptotic, and regenerative actions of MPGT make it a potential nutrient phytoceutical to rescue the stomach from alcoholic gastritis.

Keyword

Ethanol; Gastric damages; Artemisia extracts; Green tea extracts; HSP27

MeSH Terms

Alcoholics*
Artemisia*
Chronic Disease
Cyclooxygenase 2
Ethanol
Gastritis*
Heat-Shock Proteins*
Helicobacter
HSP27 Heat-Shock Proteins*
Humans
In Situ Nick-End Labeling
Oxidoreductases
Phospholipases A2
Rats, Wistar
Stomach
Stomach Diseases
Tea*
Ulcer
Cyclooxygenase 2
Ethanol
HSP27 Heat-Shock Proteins
Heat-Shock Proteins
Oxidoreductases
Phospholipases A2
Tea

Figure

• 

  Fig. 1. MPGT pretreatment alleviated ethanol-induced gastric damage. (A) Experimental scheme. Five animal groups were set: Group 2 as the control and Groups 3, 4, and 5 pretreated with 100 mg/kg, 300 mg/kg, and 500 mg/kg MPGT. The rats were sacrificed after 1 hour of 100% ethanol. Representative pictures of the gross appearance of the rat stomach after alcohol ingestion in the absence or presence of 100, 300, and 500 mg/kg MPGT pretreatment. (B) Gross and microscopic photograph of ethanol-induced gastric mucosal lesions according to the group, ×40 magnification. (C, D) The scoring criteria are written in Methods. The mean score for the gross and pathologic lesion scores. The mean levels of the gross lesion index and mean scores of pathological evaluation are presented as the mean±SD from 8 rats/group. MPGT, extracts of artemisia and green tea; SD, standard deviation.

• 

  Fig. 2. Status of ethanol-induced gastric inflammation. (A) Mean score for inflammation according to group. (B) Changes in IL-8, phosphate ERK, phosphate JNK, and phosphate p38 mRNA and their protein expression. (C) Immunohistochemistry staining with the NF-κ B p65 anti-body, ×100 magnification, and the mean positivity according to the group. ERK, extracellular signal-regulated kinases; JNK, c-Jun N-terminal kinases; mRNA, messenger RNA; SD, standard deviation.

• 

  Fig. 3. Status of ethanol-induced mucosal and submucosal damage. (A) Score of mucosal and submucosal damage. Pathology image representing the mucosal and submucosal condition from Groups 2 and 5 ×100 magnification. (B) Western blot for cPLA2 and 15-PGDH (C) RT-PCR and Western blot for COX-2 according group (left) (D) Immunohistochemical staining with COX-2 antibody (middle), ×100 magnification, and mean COX-2 positivity. cPLA2, calcium-dependent phospholipase A2; COX-2, cyclooxygenase-2; ERK, extracellular signal-regulated kinases; JNK, c-Jun N-terminal kinases, SD, standard deviation.

• 

  Fig. 4. Status of an ethanol-induced gastric ulcer. (A) Mean score for ethanol-induced gastric ulcers. The arrow shows the significant denudation and exfoliation of gastric mucosa. (B) TUNEL staining for apoptotic cells. All images were obtained from the non-ulcerated mucosa of each group and significantly increased TUNEL positive cells were noted in Group 2 even in the non-ulcerated gastric mucosa. (C) Mean TUNEL positive (apoptotic index) according to the group. (D) Western blot for BAX expression. TUNEL, terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling; SD, standard deviation.

• 

  Fig. 5. Western blot for HSP27 and HSP. (A) Changes in HSP27 and HSP70 according to group. (B) Changes in Hsp27 and Hsp70 mRNA by RT-PCT. (C) Changes in Pdgf mRNA by RT-PCR. HSP, heat-shock protein; mRNA, messenger RNA; SD, standard deviation.

Reference

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