Effects of Broccoli Seed Extract on Helicobacter pylori

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• 1.Introduction
  Helicobacter pylori (Helicobacter pylori, HP) is a bacteria that colonizes the stomach and infects more than half of the world's population. HP infection is closely related to diseases such as gastritis, peptic ulcer, gastric cancer and gastric MALT lymphoma, and is clearly defined as a Class I carcinogen of gastric cancer by the WHO. As the antibiotic resistance of HP continues to increase, leading to a decline in the eradication rate. Researchers are constantly exploring new methods and new ways of anti-HP treatment. Sulforaphane is a natural active product, widely present in cruciferous vegetables such as broccoli in the form of glucoraphanin. Studies have found that sulforaphane exhibits anti-HP activity in vivo and in vitro, regardless of whether the HP strain has drug resistance. Therefore, sulforaphane is considered to be a potential anti-HP phytochemical discovered in recent years. The natural sulforaphane against HP infection will be an important supplement to traditional triple and quadruple therapy. It is of great significance for improving the eradication rate of HP, reducing the risk of gastric cancer, and reducing the economic burden of family and society [1].
  2.Researches
  2.1 In vitro studies
  HP infection is difficult to eradicate, and treatment failure is mainly due to the emergence of drug-resistant strains, which persist in the gastrointestinal tract in the form of non-growth tolerance or as intracellular pathogens. In 2002, Fahey et al. studied the antibacterial effect of sulforaphane on HP [2]. Detected the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 3 reference strains of sulforaphane extracted from broccoli seeds and 45 HP clinical isolated from patients with gastritis and gastric or duodenal ulcer. The results showed that sulforaphane has high antibacterial activity against both sensitive and resistant strains (average MIC = 2µg/mL), indicating that the antibacterial activity of sulforaphane does not depend on the bacterial phenotype. In addition, 24 hours after the intervention of sulforaphane, it can clear the intracellular HP that causes persistent HP infection in human epithelial cells (HEp-2). This result has great clinical value, because as mentioned earlier, one of the reasons for the failure of clinical treatment is the emergence of resistant strains as intracellular pathogens.
  2.2 Animal experiments
  The antibacterial effect of sulforaphane on HP has also been confirmed by in vivo experiments. Studies by Haristoy et al. confirmed that sulforaphane can completely eradicate human gastric HP xenografts implanted in nude mice. Among the 11 xenograft nude mice treated with sulforaphane, 8 mice showed that HP infection was completely eradicated. HP-infected mice on a high-salt diet (7.5% sodium chloride), oral administration of broccoli sprouts rich in sulforaphane can reduce bacterial colonization in the stomach and reduce mucosal tumor necrosis factor-α and interleukin-1β express, reduce gastric inflammation, and prevent gastric atrophy induced by high-salt diet [3].
  2.3 Clinical researches
  Human experiments have also confirmed that the intervention of HP-infected patients with broccoli sprouts rich in sulforaphane can significantly inhibit the activity of HP and reduce the gastritis caused by HP.
  In a preliminary study on whether oral broccoli sprouts can eradicate HP infection, nine patients with HP infection consumed broccoli sprouts (14 g, 28 g or 56 g, respectively twice a day, which is equivalent to SFN 22.5 , 45 or 90 mg) for 7 days. The results showed that among the 9 patients, 7 patients turned negative on the 8th day, and 6 of them were still negative on the 35th day. Then a carbon-14 urea breath test (UBT) was performed on these 6 patients, and it was found that 2 were negative, 2 were positive, and 2 were indeterminate. One of the two patients with uncertain results agreed to tissue biopsy, and the results proved to be negative. Therefore, in general, HP was eradicated in at least 3 of the 9 patients [4].
  In a human study in 2009, 48 patients infected with HP were randomly divided into a test group and a placebo group. The test group consumed broccoli sprouts rich in sulforaphane for 8 weeks (70 g/day), and the placebo group consumed alfalfa sprouts that do not contain sulforaphane. The urea breath test and HP stool antigen (both are biomarkers of HP colonization) and serum pepsinogen I and II (biomarkers of gastric inflammation) confirmed that the level of urease was reduced. However, once the intervention is stopped, the HP level will return to the baseline level after 8 weeks [5].
  In another clinical trial in 2014, 86 patients with type 2 diabetes who were infected with HP were given standard triple therapy (20mg omeprazole, 500mg clarithromycin, 1000mg amoxicillin, twice a day for 14 days), broccoli sprout extract rich in sulforaphane (6 g/day for 28 days), or a combination of the two. The results showed that the eradication rate of sulforaphane combined with triple therapy group (91.7%) was significantly higher than that of triple therapy group alone (89.3%) and broccoli sprout group (56.0%), indicating that sulforaphane enhanced the clearance effect of standard triple therapy on HP [6].
  In summary, sulforaphane has been proven to have an inhibitory effect on HP in vitro, in vivo (including animals and humans), intracellular and extracellular HP, and can eradicate HP-resistant bacteria. Oral mucositis is a disease of mouth and throat pain caused by cancer treatment, which is often accompanied by constipation and oral odor. Our client is developing a drug for treating mucositis. According to feedback, broccoli seed extract glucoraphanin has been found to be effective in treating mucositis. The National Cancer Institute's clinical trial (NCT02023931) also proved the chemopreventive ability of broccoli seed preparations rich in sulforaphane on the carcinogenic effects of the oral environment. Another invention patent showed (CN 103717225 B) that the combination of zinc salt and isothiocyanate (sulforaphane is an isothiocyanate) can effectively reduce oral volatile sulfur compounds and thereby reduce oral odor. Therefore, we recommend the water extract of broccoli seeds (glucoraphanin and sulforaphane) as the core raw materials to intervene in adjusting the harmful oral flora and eliminate odors.
  References
  【1】Moeller H, Correa P. Carcinogenicity of some biological agents. Cancer Epidemiol Biomarkers Prev. 1994 Oct-Nov;3(7):627.
  【2】Fahey JW, Haristoy X, Dolan PM, et al. Sulforaphane inhibits extracellular, intracellular, and antibiotic-resistant strains of Helicobacter pylori and prevents benzo[a]pyrene-induced stomach tumors. Proc Natl Acad Sci U S A. 2002 May 28;99(11):7610-5.
  【3】Haristoy X, Angioi-Duprez K, Duprez A, Lozniewski A. Efficacy of sulforaphane in eradicating Helicobacter pylori in human gastric xenografts implanted in nude mice. Antimicrob Agents Chemother. 2003 Dec;47(12):3982-4.
  【4】Galan MV, Kishan AA, Silverman AL. Oral broccoli sprouts for the treatment of Helicobacter pylori infection: a preliminary report. Dig Dis Sci. 2004 Aug;49(7-8):1088-90. 
  【6】Yanaka A, Fahey JW, Fukumoto A, et al. Dietary sulforaphane-rich broccoli sprouts reduce colonization and attenuate gastritis in Helicobacter pylori-infected mice and humans. Cancer Prev Res (Phila). 2009 Apr;2(4):353-60.
  【6】Mirmiran P, Bahadoran Z, Golzarand M, et al. A comparative study of broccoli sprouts powder and standard triple therapy on cardiovascular risk factors following H.pylori eradication: a randomized clinical trial in patients with type 2 diabetes. J Diabetes Metab Disord. 2014 May 28;13:64.