BEE VENOM FOR OBESITY
What Is Obesity?
It is a medical condition, which arises when a person has additional weight in the form of stored fat that can affect overall health. Obesity can be termed as a complex disease, as it can complicate into different issues like certain cancers, high blood pressure, diabetes, and heart disease. So it is more than just a cosmetic factor.
For many people, it is very difficult to avoid obesity and that’s due to an inherited factor combined with poor diet and an inactive lifestyle. For other subjects of obesity, without any hereditary reason, behavior changes increased physical activity, dietary changes and certain medications can help lose weight. Weight loss regime along with prescription medicines are usually meant for people having chronic obesity.
How Is Obesity Detected?
Normally, the BMI tool is used to assess the weight of a person according to sex, age, weight, and height. BMI index above 25 indicates that a person is overweight, while above 30 indicates that a person is obese.
A person's body shape and weight are further determined through the distribution of fat, waist to height and waist to hip size ratio. An obese person can also develop arthritis and metabolic syndrome. Under metabolic syndrome come different issues like type II diabetes and high blood pressure.
What Factors Lead To Obesity?Several factors can lead to obesity in men and women and they are stated below:
How Is Obesity Treated?
A professional doctor will conduct some tests after a physical examination. It is important to take into regard the health history of an obese person and then make an appropriate diagnosis. This will eventually make the treatment process a lot easier and effective, in the long run.
Many doctors recommend strong medicines to obese people, which help with weight loss, but damage other organs like kidneys and liver. These medicines work unnaturally to help a person get rid of excess fat. What an obese person needs to do is; start exercising daily and have a healthy diet and then choose a natural medicine that will help get rid of accumulated fat.
Bee venom contains various essential components that are useful in different medical conditions. It has been observed that bee venom with a proper dose regime can help get rid of obesity. And the best part is that, when administered carefully, there are no side effects of this treatment.
Bee Venom Suppresses the Differentiation of Preadipocytes and High Fat Diet-Induced Obesity by Inhibiting Adipogenesis
Introduction
Obesity results due to excessive or abnormal accumulation of fat. Hypertrophy and adipocyte hyperplasia are determinant factors of this medical condition. Adipocytes differentiate from other precursor cells or stem cells and that is a complex mechanism of the gene expression that results in different obesity-related diseases. 3T3-L1 preadipocyte, an undifferentiated fibroblast, has been used in different clinical studies; related to differentiation and adipogenesis. These cells have a different repose to the adipogenic inducers that include dexamethasone, IBMX (3-isobutyl-1-methylxanthine) and insulin.
Differentiation sequence from the preadipocytes to adipocytes consists of terminal differentiation, mitotic clonal expansion, and confluence. What happens in the first stage is that, the confluent cells become a part of the growth arrest phase. Later, these cells restart their cycle and enhance the cell numbers 3 to 4 times during the MCE phase. Hyperplasia experienced during this cell differentiation phase is linked with the production of adipogenic transcription factors.
It is noted that CCAAT/ (C/EBPs) and PPAR (peroxisome proliferator-activated receptors) γ promote the differentiation phase of adipocytes. In an early stage, differentiation of the 3T3-L1 Cells, expression of C/EBPδ and C/EBPβ were significantly increased after a hormonal induction that ultimately increases expression of PPARγ and C/EBPα. C/EBPδ is very important for MCE to originate during the differentiation phase in an early stage of adipogenesis. The gene expression of C/EBPβ persuades the expression of C/EBPα and PPARγ. Complete activation of PPARγ and C/EBP family regulates the expression of different adipogenic factors that support fat accumulation.
AMPK (adenosine monophosphate-activated protein kinase), is also known as a prime controller of energy homeostasis, serves as an important target for managing obesity. During adipogenesis, activation of AMPK regulates lipid and glucose metabolism through the inactivation of metabolic enzymes. AMPK's phosphorylation inactivates ACC (acetyl coenzyme which is a carboxylase) and HMGCR (3-hydroxy-3-methylglutaryl coenzyme which is a reductase), which leads to prevention of cholesterol and fatty acids synthesis and increased oxidation of fatty acid. It has also been found that AMPK controls channeling of acyl-CoA towards lipid biosynthesis and beta-oxidation that leads to inhibition of GPAT (glycerol-3-phosphate acyltransferase). Furthermore, AMPKs phosphorylation also prevents the expression of adipogenic transcription factors like PPARγ, C/EBPα, C/EBPδ, and C/EBPβ.
BV (bee venom) is a complex of different peptides, proteins and other low weight components that act as an effective defense tool. In spite of the painful sting of a honeybee, its venom has been used for a long time, in different forms, to treat pain, diseases, and even tumors. It has been found that inhibition of the atherosclerotic lesions through suppression of non-harmful prostatic hyperplasia and anti-inflammatory mechanism in clinical rats is presented by bee venom.
Melittin is perhaps the most useful component of bee venom, which is responsible for the downregulation of pro-inflammatory cytokines, NF-κB signal pathway, proatherogenic proteins, and adhesion molecules and for improvement in the atherosclerotic lesion in high fat treated experimental animal models. Adding more, macrophage infiltration, fibrotic factors, adhesion molecules, pro-inflammatory cytokines, and apamin attenuated lipids in the fat-induced atherosclerotic experimental mice was observed. It was reported that bee venom exhibits anti-obesity effects. The mechanism behind it needs to be studied carefully. In this clinical experiment anti-obesity effects of bee venom in the 3T3-L1 preadipocytes in HFD induced obesity models were investigated carefully.
Method
Oil red O, insulin, DEX, IBMX and BVA, and other chemicals were obtained from Sigma Chemical, USA. Penicillin-streptomycin, fetal bovine serum, bovine serum, and Dulbecco's modified eagles medium were all obtained from Life Technologies, USA. While antibodies against β-actin (C4; cat. no. sc-47778), C/EBPα (C-18; cat. no. sc-9314) and PPARγ (E-8; cat. no. sc-7273) were obtained from the Santa Cruz Biotechnology, Inc, USA. ERK, JNK, phosphor stress-activated protein kinase/Jun-amino terminal kinase, Phospho extracellular signal-regulated kinase, phospho-p38 MAPK, p38, AMPK, p-AMPK, ACC and p-ACC antibodies were acquired from Cell Signaling Technology, USA, while the horseradish peroxidase-conjugated antibodies were acquired from the Jackson ImmunoResearch Laboratories, Inc. The USA. The SYBR Green Master Mix was acquired from the Applied Biosystems, USA. PPARγ, C/EBPδ, C/EBPα, C/EBPβ, and GAPDH were obtained from Bioneer, Korea.
3T3-L1, Preadipocytes were obtained from Korean Cell Line Band, Korea and then cultured in DMEM with a supplement of 10% of BS, 100µg/mL of streptomycin and 100U/mL of penicillin in an incubator having a temperature of 37 degrees centigrade and 5% carbon dioxide. Adipocyte differentiation was conducted through culturing of 3T3-L1 cells in dishes at a density level of 2x105 cells/mL to confluence. During the stage of full confluence, the plates were treated with different concentrations of bee venom. During the second stage, which started on the day 3 of differentiation, cells were subjected to 1µ/mL of insulin in the DMEM with a 10% FBS (v/v) and 1% of PS. During the third or final stage, cells were shifted to the DMEM having 1%PS and 10%FBS – culture media was replaced after 3 days.
Viability of cells was assessed through an MTT (3 - (4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) assay. 3T3-L1 cells were placed in a 96 well plate and were treated with different concentrations of bee venom for some time of 72hours at a temperature of 37 degrees centigrade having some humidity and 5% of carbon dioxide. Once the treatment was completed, cells were subjected to staining with an MTT solution for 4hours at a temperature of 37 degrees centigrade. Once the excess reagent was removed, the insoluble formazan was dissolved in DMSO. Cell viability was calculated at 570nm through the Epoch microvolume spectrophotometer.
Total RNA was subjected to isolation and the cells were homogenized with a Trizol reagent obtained from Invitrogen, USA. The cDNA was acquired through isolation of total RNA, a d 16 primer and AMV-RT. Relative gene expression was calculated through real-time PCR using the SYBR green PCR master mix. Gene Ct values of C/EBPδ, PPARγ, C/EBPα, and C/EBPβ were normalized through a Gene Express Program.
Acquired blood samples were centrifuged at 1003xg for 15 minutes at normal temperature to generate serum samples. They were later stored at -80 degrees centigrade to conduct further measurements. Serum concentrations of LDL cholesterol and triglyceride were determined through enzymatic methods conducted with commercial kits from BioVision, USA. Other processes involved were: MTT assay, Oil Red O staining, western blot analysis, histological analysis, and statistical analysis.
Results
It is known that lipid and hyperplasia accumulation occurs in the 3T3-L1 cell line during the process of differentiation. These cell lines were treated with bee venom having concentration starting from 1.25µg/mL and going up till 40µg/mL in MDO or BS. After treating cell lines with bee venom, cell viability was assessed by an MTT (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) assay. It was found that bee venom had no sort of effect on cell viability in culture media but bee venom reduced cell viability at 2.5µg/mL and higher concentrations.
Inhibitory effect of bee venom on lipid accumulation in the adipocytes treated with and without bee venom for 9 days was determined. It was noticed that the differentiation induced adipocytes dramatically increased lipid storage by nearly 2 times when compared with the undifferentiated cells. On the other hand, bee venom treatment impressively reduced the lipid droplet accumulation, but in a dose-dependent system.
Transcriptions factors like the C/EBP family and PPARγ play an important role in the differentiation and maturation of adipocytes. The anti adipogenesis effect of bee venom in the differentiated adipocytes was investigated through Western blot analysis and qRT-PCR. It was established that the mRNA expression of PPARγ, C/EBPδ, C/EBPα, and C/EBPβ was upregulated during differentiation induced adipocytes when compared to what was observed in the undifferentiated cells. Bee venom treatment helped in downregulating mRNA expression of the C/EBP family and also decreased the PPARγ expression for both protein and mRNA levels.
To examine the effect of bee venom on the MAPK pathway, protein expression of different factors involved in the MAPK pathway was determined through Western blot analysis. This process was done in differentiated adipocytes. It was noticed that the phosphorylation of JNK and ERK decreased while phosphorylation of p38 went up during differentiation when compared to the results of undifferentiated control cells. Phosphorylation of JNK and ERK was significantly upregulated in the bee venom treated adipocytes while phosphorylation of p38 was remarkably decreased.
This clinical study also revealed that the overall bodyweight of mice belonging to the HFD (high-fat diet) group increased when compared to the bodyweight of the ND (normal diet) group. Totally in contrast to the HFD group, bee venom injection decreased the overall body weight, weight gain, and fat. Obesity is generally characterized by hyperplasia and the hypertrophy of adipose tissue and for that reason, inhibitory effects of bee venom were examined through H and E staining. H and E staining analysis data showed that the HFD group of mice indicated hypertrophy of the adipocytes in epididymal adipose tissue, while the bee venom treated group demonstrated repressed hypertrophy of the adipocytes.
Conclusion
In the end, we can declare that this clinical study successfully established that bee venom prevents an early adipogenic process by downregulating the MCE stage through regulation of PPARγ, AMPK, ERK, and C/EBPs signaling. According to these findings, it can also be said that bee venom can be used as an effective therapeutic and preventive agent in treating obesity.
Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5793096/
It is a medical condition, which arises when a person has additional weight in the form of stored fat that can affect overall health. Obesity can be termed as a complex disease, as it can complicate into different issues like certain cancers, high blood pressure, diabetes, and heart disease. So it is more than just a cosmetic factor.
For many people, it is very difficult to avoid obesity and that’s due to an inherited factor combined with poor diet and an inactive lifestyle. For other subjects of obesity, without any hereditary reason, behavior changes increased physical activity, dietary changes and certain medications can help lose weight. Weight loss regime along with prescription medicines are usually meant for people having chronic obesity.
How Is Obesity Detected?
Normally, the BMI tool is used to assess the weight of a person according to sex, age, weight, and height. BMI index above 25 indicates that a person is overweight, while above 30 indicates that a person is obese.
A person's body shape and weight are further determined through the distribution of fat, waist to height and waist to hip size ratio. An obese person can also develop arthritis and metabolic syndrome. Under metabolic syndrome come different issues like type II diabetes and high blood pressure.
What Factors Lead To Obesity?Several factors can lead to obesity in men and women and they are stated below:
- Consuming additional calories for a long period
- Leading an inactive lifestyle
- Not getting enough sleep
- Presence of endocrine disruptors
- Weight gain due to any certain medication
- Obesity gene
How Is Obesity Treated?
A professional doctor will conduct some tests after a physical examination. It is important to take into regard the health history of an obese person and then make an appropriate diagnosis. This will eventually make the treatment process a lot easier and effective, in the long run.
Many doctors recommend strong medicines to obese people, which help with weight loss, but damage other organs like kidneys and liver. These medicines work unnaturally to help a person get rid of excess fat. What an obese person needs to do is; start exercising daily and have a healthy diet and then choose a natural medicine that will help get rid of accumulated fat.
Bee venom contains various essential components that are useful in different medical conditions. It has been observed that bee venom with a proper dose regime can help get rid of obesity. And the best part is that, when administered carefully, there are no side effects of this treatment.
Bee Venom Suppresses the Differentiation of Preadipocytes and High Fat Diet-Induced Obesity by Inhibiting Adipogenesis
Introduction
Obesity results due to excessive or abnormal accumulation of fat. Hypertrophy and adipocyte hyperplasia are determinant factors of this medical condition. Adipocytes differentiate from other precursor cells or stem cells and that is a complex mechanism of the gene expression that results in different obesity-related diseases. 3T3-L1 preadipocyte, an undifferentiated fibroblast, has been used in different clinical studies; related to differentiation and adipogenesis. These cells have a different repose to the adipogenic inducers that include dexamethasone, IBMX (3-isobutyl-1-methylxanthine) and insulin.
Differentiation sequence from the preadipocytes to adipocytes consists of terminal differentiation, mitotic clonal expansion, and confluence. What happens in the first stage is that, the confluent cells become a part of the growth arrest phase. Later, these cells restart their cycle and enhance the cell numbers 3 to 4 times during the MCE phase. Hyperplasia experienced during this cell differentiation phase is linked with the production of adipogenic transcription factors.
It is noted that CCAAT/ (C/EBPs) and PPAR (peroxisome proliferator-activated receptors) γ promote the differentiation phase of adipocytes. In an early stage, differentiation of the 3T3-L1 Cells, expression of C/EBPδ and C/EBPβ were significantly increased after a hormonal induction that ultimately increases expression of PPARγ and C/EBPα. C/EBPδ is very important for MCE to originate during the differentiation phase in an early stage of adipogenesis. The gene expression of C/EBPβ persuades the expression of C/EBPα and PPARγ. Complete activation of PPARγ and C/EBP family regulates the expression of different adipogenic factors that support fat accumulation.
AMPK (adenosine monophosphate-activated protein kinase), is also known as a prime controller of energy homeostasis, serves as an important target for managing obesity. During adipogenesis, activation of AMPK regulates lipid and glucose metabolism through the inactivation of metabolic enzymes. AMPK's phosphorylation inactivates ACC (acetyl coenzyme which is a carboxylase) and HMGCR (3-hydroxy-3-methylglutaryl coenzyme which is a reductase), which leads to prevention of cholesterol and fatty acids synthesis and increased oxidation of fatty acid. It has also been found that AMPK controls channeling of acyl-CoA towards lipid biosynthesis and beta-oxidation that leads to inhibition of GPAT (glycerol-3-phosphate acyltransferase). Furthermore, AMPKs phosphorylation also prevents the expression of adipogenic transcription factors like PPARγ, C/EBPα, C/EBPδ, and C/EBPβ.
BV (bee venom) is a complex of different peptides, proteins and other low weight components that act as an effective defense tool. In spite of the painful sting of a honeybee, its venom has been used for a long time, in different forms, to treat pain, diseases, and even tumors. It has been found that inhibition of the atherosclerotic lesions through suppression of non-harmful prostatic hyperplasia and anti-inflammatory mechanism in clinical rats is presented by bee venom.
Melittin is perhaps the most useful component of bee venom, which is responsible for the downregulation of pro-inflammatory cytokines, NF-κB signal pathway, proatherogenic proteins, and adhesion molecules and for improvement in the atherosclerotic lesion in high fat treated experimental animal models. Adding more, macrophage infiltration, fibrotic factors, adhesion molecules, pro-inflammatory cytokines, and apamin attenuated lipids in the fat-induced atherosclerotic experimental mice was observed. It was reported that bee venom exhibits anti-obesity effects. The mechanism behind it needs to be studied carefully. In this clinical experiment anti-obesity effects of bee venom in the 3T3-L1 preadipocytes in HFD induced obesity models were investigated carefully.
Method
Oil red O, insulin, DEX, IBMX and BVA, and other chemicals were obtained from Sigma Chemical, USA. Penicillin-streptomycin, fetal bovine serum, bovine serum, and Dulbecco's modified eagles medium were all obtained from Life Technologies, USA. While antibodies against β-actin (C4; cat. no. sc-47778), C/EBPα (C-18; cat. no. sc-9314) and PPARγ (E-8; cat. no. sc-7273) were obtained from the Santa Cruz Biotechnology, Inc, USA. ERK, JNK, phosphor stress-activated protein kinase/Jun-amino terminal kinase, Phospho extracellular signal-regulated kinase, phospho-p38 MAPK, p38, AMPK, p-AMPK, ACC and p-ACC antibodies were acquired from Cell Signaling Technology, USA, while the horseradish peroxidase-conjugated antibodies were acquired from the Jackson ImmunoResearch Laboratories, Inc. The USA. The SYBR Green Master Mix was acquired from the Applied Biosystems, USA. PPARγ, C/EBPδ, C/EBPα, C/EBPβ, and GAPDH were obtained from Bioneer, Korea.
3T3-L1, Preadipocytes were obtained from Korean Cell Line Band, Korea and then cultured in DMEM with a supplement of 10% of BS, 100µg/mL of streptomycin and 100U/mL of penicillin in an incubator having a temperature of 37 degrees centigrade and 5% carbon dioxide. Adipocyte differentiation was conducted through culturing of 3T3-L1 cells in dishes at a density level of 2x105 cells/mL to confluence. During the stage of full confluence, the plates were treated with different concentrations of bee venom. During the second stage, which started on the day 3 of differentiation, cells were subjected to 1µ/mL of insulin in the DMEM with a 10% FBS (v/v) and 1% of PS. During the third or final stage, cells were shifted to the DMEM having 1%PS and 10%FBS – culture media was replaced after 3 days.
Viability of cells was assessed through an MTT (3 - (4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) assay. 3T3-L1 cells were placed in a 96 well plate and were treated with different concentrations of bee venom for some time of 72hours at a temperature of 37 degrees centigrade having some humidity and 5% of carbon dioxide. Once the treatment was completed, cells were subjected to staining with an MTT solution for 4hours at a temperature of 37 degrees centigrade. Once the excess reagent was removed, the insoluble formazan was dissolved in DMSO. Cell viability was calculated at 570nm through the Epoch microvolume spectrophotometer.
Total RNA was subjected to isolation and the cells were homogenized with a Trizol reagent obtained from Invitrogen, USA. The cDNA was acquired through isolation of total RNA, a d 16 primer and AMV-RT. Relative gene expression was calculated through real-time PCR using the SYBR green PCR master mix. Gene Ct values of C/EBPδ, PPARγ, C/EBPα, and C/EBPβ were normalized through a Gene Express Program.
Acquired blood samples were centrifuged at 1003xg for 15 minutes at normal temperature to generate serum samples. They were later stored at -80 degrees centigrade to conduct further measurements. Serum concentrations of LDL cholesterol and triglyceride were determined through enzymatic methods conducted with commercial kits from BioVision, USA. Other processes involved were: MTT assay, Oil Red O staining, western blot analysis, histological analysis, and statistical analysis.
Results
It is known that lipid and hyperplasia accumulation occurs in the 3T3-L1 cell line during the process of differentiation. These cell lines were treated with bee venom having concentration starting from 1.25µg/mL and going up till 40µg/mL in MDO or BS. After treating cell lines with bee venom, cell viability was assessed by an MTT (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) assay. It was found that bee venom had no sort of effect on cell viability in culture media but bee venom reduced cell viability at 2.5µg/mL and higher concentrations.
Inhibitory effect of bee venom on lipid accumulation in the adipocytes treated with and without bee venom for 9 days was determined. It was noticed that the differentiation induced adipocytes dramatically increased lipid storage by nearly 2 times when compared with the undifferentiated cells. On the other hand, bee venom treatment impressively reduced the lipid droplet accumulation, but in a dose-dependent system.
Transcriptions factors like the C/EBP family and PPARγ play an important role in the differentiation and maturation of adipocytes. The anti adipogenesis effect of bee venom in the differentiated adipocytes was investigated through Western blot analysis and qRT-PCR. It was established that the mRNA expression of PPARγ, C/EBPδ, C/EBPα, and C/EBPβ was upregulated during differentiation induced adipocytes when compared to what was observed in the undifferentiated cells. Bee venom treatment helped in downregulating mRNA expression of the C/EBP family and also decreased the PPARγ expression for both protein and mRNA levels.
To examine the effect of bee venom on the MAPK pathway, protein expression of different factors involved in the MAPK pathway was determined through Western blot analysis. This process was done in differentiated adipocytes. It was noticed that the phosphorylation of JNK and ERK decreased while phosphorylation of p38 went up during differentiation when compared to the results of undifferentiated control cells. Phosphorylation of JNK and ERK was significantly upregulated in the bee venom treated adipocytes while phosphorylation of p38 was remarkably decreased.
This clinical study also revealed that the overall bodyweight of mice belonging to the HFD (high-fat diet) group increased when compared to the bodyweight of the ND (normal diet) group. Totally in contrast to the HFD group, bee venom injection decreased the overall body weight, weight gain, and fat. Obesity is generally characterized by hyperplasia and the hypertrophy of adipose tissue and for that reason, inhibitory effects of bee venom were examined through H and E staining. H and E staining analysis data showed that the HFD group of mice indicated hypertrophy of the adipocytes in epididymal adipose tissue, while the bee venom treated group demonstrated repressed hypertrophy of the adipocytes.
Conclusion
In the end, we can declare that this clinical study successfully established that bee venom prevents an early adipogenic process by downregulating the MCE stage through regulation of PPARγ, AMPK, ERK, and C/EBPs signaling. According to these findings, it can also be said that bee venom can be used as an effective therapeutic and preventive agent in treating obesity.
Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5793096/
