RESEARCH OF A. LAPPA L. LEAVES MICROBIAL-FERMENTED BIOMASS HYPOGLYCEMIC ACTIVITY

RESEARCH OF A. LAPPA L. LEAVES MICROBIAL-FERMENTED BIOMASS HYPOGLYCEMIC ACTIVITY

Ruslan Khaibullin

Postgraduate student of «Chemistry and biotechnology» department, Perm National Research Polytechnic University (PNRPU),

Russia, Perm

Anna Burluckaya

Researcher of Pharmacology department, Perm State Medical University (PSMU),

Russia, Perm

Larisa Volkova

Doctor of Medicine, Professor of «Chemistry and biotechnology» department, Perm National Research Polytechnic University (PNRPU),

Russia, Perm

Galina Vdovina

Doctor of Pharmacy, Professor of Pharmacology department, Perm State Medical University (PSMU),

Russia, Perm

ABSTRACT

Arctium lappa L. (burdock) leaves fermented biomass is a promising substance for the pharmaceutical preparations production for the metabolic syndrome correction and prevention due to the presence of inulin polysaccharide, as well as bacterial metabolites exhibiting biologically active properties.

The article presents experimental data on the hypoglycemic effect of A. lappa L fermented leaf biomass extracts on laboratory animals. The fermented biomass hypoglycemic activity was found at a 500 mg/kg of animal body weight concentration.

Keywords: A. lappa L. leaves fermented biomass, blood sugar level, hypoglycemic effect, metabolic syndrome.

Introduction

Metabolic syndrome (MS) is a body pathologies combination that increase the risk of arterial hypertension, cardiovascular diseases, atherosclerosis, and type 2 diabetes mellitus (DM2) [1]. An important role in the prevention and treatment of MS and DM2 is played by medicinal plants, one of which is widely used A. lappa L. root biomass [2,3]. A. lappa L. leaf biomass fermentation with the help of Lactobacillus genus microorganisms allows not only to expand the traditional use of this plant's roots, but also to enrich the resulting product with bacterial metabolites, which have an additional positive effect on the gastrointestinal tract state [4, 5]. In combination with polysaccharides, in particular inulin, which are part of plant materials, enrichment with the probiotics metabolites can make it possible to use this fermented product as a means for the prevention and adjuvant therapy of metabolic disorders.

Thus, the assessment of the A. lappa L. leaves fermented pharmaceutical substance hypoglycemic activity is an urgent task.

Materials and methods

In the experiment, water extracts of the A. lappa L. leaves biomass fermented substances were studied. This fermented substances was obtained by the previously developed microbial fermentation technology [6]. For the research, 3 test substance's concentrations, based on the main active substance - inulin: 100/60/20 mg of inulin per kg of animal body weight or, respectively, 500/300/100 mg of the substance per kg of body weight were used. Metformin (Siofor®500, MENARINI-VON HEYDEN, Germany) was used as a reference drug.

The evaluation of the obtained substances hypoglycemic properties was carried out on laboratory animals according to the glucose tolerance test method [7]. In the experiment, male healthy white outbred rats with an average weight of 315.0 ± 10.4 g were used in the amount of 40 pieces, divided into 5 groups of 8 animals in each:

1) Control group;

2) The group that received metformin;

3) The group that received the fermented substance's aqueous extract at the rate of 500 mg of the substance per 1 kg of animal body weight;

4) The group that received the fermented substance's aqueous extract at the rate of 300 mg of the substance per 1 kg of animal body weight;

5) The group that received the fermented substance's aqueous extract at the rate of 100 mg of the substance per 1 kg of animal body weight.

Previously, all animals were subjected to 12-hour food deprivation with free access to water. Further, 40% glucose solution at a dosage of 4 ml/kg of body weight was administered once orally into the animals' organism. After that, oral administration was carried out in amounts of 7.5 ml of extract per 1 kg of body weight:

1. Group № 1 - water;
2. Group № 2 - metformin solution (Siofor®500, MENARINI-VON HEYDEN, Germany) 85 mg/kg;
3. Group № 3 - fermented substance extract 500mg/kg;
4. Group № 4 - fermented substance extract 300mg/kg;
5. Group № 5 - fermented substance extract 100mg/kg.

The blood glucose level was assessed using an Accu-Chek Active glucometer (Roche Diagnostics, Switzerland). Blood was taken from the fingers of the animal hind limb. Blood glucose concentrations were measured on an empty stomach before the administration of glucose, then 30, 60, and 120 minutes after the administration of glucose, metformin, and extracts of the test substance. The glycemic level in the dynamics of mmol/L was assessed using descriptive statistics methods.

The matrix of the experiment is presented in table 1 (table 1).

Table 1.

Matrix for planning an assess hypoglycemic activity experiment

Experimental part

The experiment results are presented in table 2 (table 2).

Table 2.

The results of the hypoglycemic activity assessment

According to the results obtained, it was found that the highest glycemic level (blood sugar level increase from 42 to 79%) was achieved 60 minutes after the experiment start in the control group case and 30 minutes after the experiment start in the experimental groups case, then began the decrease in blood glucose concentration.

The results obtained, in accordance with the Student's t-test statistical evaluation data, show the significance of the metformin hypoglycemic activity at a concentration of 85 mg/kg in the range from 60 to 120 minutes and the resulting substance at a concentration of 500 mg/kg in the range of 60 minutes compared with the control group. The dynamics of glycemia is shown in Figure 1 (Figure 1).

Figure 1. Test substance glycemic dynamics over time

Conclusion

Thus, in the resulting A. lappa L. fermented pharmaceutical substance at a 500 mg/kg of body weight concentration, when conducting a glucose tolerance test on laboratory animals, a statistically significant hypoglycemic activity was found compared to the control group.

It should be noted that the hypoglycemic activity of preparations based on A. lappa L. biomass was previously studied in the long term. This, in comparison with the positive results of a short-term study of the resulting fermented substance, indicates its effectiveness and the need for additional longer studies.

References:

1. Bastaki, S. Diabetes mellitus and its treatment // Journal of diabetes and metabolism. 13 (3). 2005. 111-134;
2. Kooti, W., Farokhipour, M., Asadzadeh, Z., Ashtary-Larky, D., Asadi-Samani, M. (2016). The role of medicinal plants in the treatment of diabetes: a systematic review // Electron Phys. 8 (1), 1832–1842. doi: 10.19082/1832;
3. Chan, Yuk-Shing at al. A review of the pharmacological effects of Arctium lappa (burdock) // Inflammopharmacology. 19. 2010. 245-54. 10.1007/s10787-010-0062-4;
4. Khaibullin R.G., Volkova L.V. Perspektivy poluchenija biologicheski aktivnyh substancij iz biomassy rastenij metodom mikrobnoj fermentacii (obzor) [Perspectives for production plant biomass biologically active product by the microbial fermentation method (review)]// Voprosy biologicheskoj, medicinskoj i farmacevticheskoj himii. [Problems of biological medical and pharmaceutical chemistry] - 2021; 4: 3−12; [in Russian]
5. Khaibullin R.G., Volkova L.V. Matematicheskoe modelirovanie nakopleniya polisaxaridov v biomasse list`ev A. lappa L. v processe mikrobnoj fermentacii [Mathematical modeling of polysaccharides accumulation in A. Lappa L. leaves biomass in microbial fermentation process]// Mezhdunarodny`j nauchno-issledovatel`skij zhurnal. [International research journal] 2022. - № 4 (118), ch. 2. - S. 178-183; [in Russian]
6. Pat. № 2733141 Russian Federation. Sposob poluchenija fitokompozicii / Volkova L.V., Khaibullin R.G.; The applicant and the patentee Perm national research polytechnic university - № 2019119049: appl. 06/18/19: publ. 29.09.20. - 6 s.;
7. Doklinicheskie issledovanija lekarstvennyh veshhestv : ucheb. posobie [Preclinical studies of medicinal substances: textbook]/ A.V. Buzlama [at al.] ; ed. by. A. A. Svistunova. - M. : GJeOTAR-Media, 2017. - 384 p. [in Russian]