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Phytochemical, hematologic and anti-tumor activity evaluations of Carica papaya leaf extract

Hoang Nguyen

California Health Solutions Services, California, USA

Department of Biological Chemistry and Pharmacology, College of Medicine, The Ohio State University, Columbus, Ohio, USA

E-mail : bhuvaneswari.bibleraaj@uhsm.nhs.uk

Joseph P Kitzmiller

Department of Biological Chemistry and Pharmacology, College of Medicine, The Ohio State University, Columbus, Ohio, USA

Pooja Khungar

Advocate Masonic, Chicago, Illinois, USA

Narendra Dabbade

Gottlieb Memorial Hospital/Loyola University Medical School, Chicago, Illinois, USA

Kieu Tho Nguyen

Gia Dinh Hospital/Ho Chi Minh University School of Medicine and Pharmacy, Ho Chi Minh City, Vietnam

Elizabeth Lynn Flynn

University of Florida, School of Pharmacy, Gainesville, Florida, USA

DOI: 10.15761/JTS.1000185

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Abstract

Carica Papaya has been used for its homeostatic and health-promoting properties in several non-Western medicinal practices (e.g., Ayurveda, Traditional Chinese and Vietnamese, Unani) for centuries. More recently, anecdotal reports have emerged suggesting regular consumption of tea made from C. Papaya leaf extract has been associated with remission of certain cancers including some advanced solid-tumor cancers. We aimed, therefore, to use scientific methodologies to investigate C. Papaya leaf extract and its in vitro effects on human cancer cells and in vivo effects in human patients with cancer. Liquid chromatography-mass spectroscopy (LC-MS) was used to determine the phytochemical composition of C. Papaya leaf extract, immune-modulatory and ant-cancer properties were studies in peripheral blood mononuclear cells and various solid-tumor cells lines, and clinical laboratory measurements (hematologic and organ-specific and non-specific parameters) were examined in 116 cancer patients receiving capsules of C. Papaya leaf extract. Our results indicate that C. Papaya may have substantial potential as an adjuvant therapy for certain cancers, and additional investigation is warranted.

Key words

Carica Papaya, papaya, liquid chromatography-mass spectroscopy, phytochemistry, hematology, anti-tumor effect, ethnopharmacology, alternative medicine

Introduction

Plants provide a healthful source of nutrition, and the use of plants for medicinal purposes dates to ancient history [1]. Contemporary consensus holds that diets high in fruits and vegetables promote good health and overall wellbeing [2]. Therapeutic properties of various plants and plant components have been demonstrated using modern clinical sciences techniques (e.g., randomized controlled trials) [3-5]. Reports of findings from studies of Carica Papaya have suggested that phytochemicals (carotenoids and polyphenols) in the leaf extract have healthful properties, promoting both anti-tumor and homeostatic activity [6-9]. Native to Central and South America and parts of southern Mexico, C. Papaya is cultivated in most tropical countries. Its leaves, fruits, latex, and roots are routinely used as a natural remedy against a variety of ailments. The constituents of the various parts of the C. Papaya plant (Table 1) have been determined and reported previously [6]. Here in we report our in vitro and in vivo investigations of C. Papaya leaf extract.

Table 1. Primary constituents of C. Papaya

Part

Constituents

Fruits

Fat, carbohydrates, phosphorous, iron, Vitamin C, thiamine, riboflavin, niacin, and carotene, amino acids, citric and malic acids (green fruits), volatile compounds: linalool, benzylisothiacyanate, protein, fat and fiber

Latex

Peptidase A and B and lysozymes, proteolytic enzymes, papain and chemopapain, glutamine cyclotransferase, chymopapains A, B and C

Leaves

Vitamin C and E, Alkaloids carpain, pseudocarpain and dehydracarpaine I and II, choline, carposide

Root

Enzyme myrosin and carposide

Materials and methods

Collection and Processing of C. Papaya

C. Papaya was purchased from a market region in Gainesville, Florida (USA) and deposited at the lab of the Department of Pharmaceutical Outcomes and Policy at the College of Pharmacy of the University of Florida (USA). C. Papaya leaf specimens were authenticated by visual inspection, rinsed thoroughly with purified water, size-reduced, and triturated to a fine powder. A uniform mortar paste was formed by combining the fine powder with minimal amounts of purified water. Gravity filtration of the paste resulted in purified C. Papaya leaf extract.

Phytochemical Analysis of C. Papaya Leaf Extract

Laser-coupled mass spectrometry (LCMS) methodology was utilized to conduct the phytochemical analysis of the purified extract of C. Papaya leaf. The extract was subjected to chromatographic separation on Phenomonex Phase C-18 Columns (water:acetonitrile ratio 80:20, 24 degrees Celsius, flow rate 1.4ml/min, electronic spray ionization mode, m/z range 50-1000) [10,11]. Spectra analysis in both positive and negative mode were performed to determine the phytochemical constituents of the purified extract of C. Papaya leaf.

In vitro Studies of C. Papaya Leaf Extract

To assess potential anti-cancer properties of C. Papaya, measurements of 3H-thymidine (3[H]- TdR) incorporation in various solid-tumor cell lines (Hela, MCF-7, HepG2, PC-14, Panc-1 and H2452) and large-cell lymphoma cell lines (Jurkat, ARH77, Raji and Karpas-299) after 24-hour exposure to various concentrations of C. Papaya leaf extract were performed [12]. Annexin V measurements in controls and after exposure to C. Papaya leaf extract at various concentrations were also compared, and measurements of luminescence after 0, 18 and 24-hour exposure to etoposide or C. Papaya leaf extract were compared. To investigate the effect of C. Papaya leaf extract on cytokine production, peripheral blood mononuclear cells (PBMCs) were stimulated with immobilized anti-CD3 and soluble CD28 in the presence or absence of C. Papaya (24-hour exposure) [13]. Supernatants from triplicate cultures were collected and quantification of IL-2, IL-4, IL-10, IL-12p40, IFN-γ, and TNF-α was performed by enzyme-linked immunosorbent assay (ELISA). The cytotoxicity of C. Papaya leaf extract was evaluated by measuring 3[H]- TdR incorporation and percent lysis in PBMC controls and in PBMCs after 24-hour exposure to C. Papaya leaf extract at various effector-target (E/T) ratios [14]. To investigate the effect of C. Papaya leaf extract on genetic expression and regulation, mRNA concentrations were measured in PBMCs without exposure and with 24-hour exposure to various doses of C. Papaya leaf extract. Quantification of mRNA was performed by real-time polymerase chain reaction (PCR) standard methodology.

Clinical Investigation of Capsules of C. Papaya Leaf Extract

Purified leaf extract of C. Papaya was dried for 20 hours to obtain low moisture and encapsulated, producing capsules of uniform C. Papaya leaf extract content suitable for human consumption. Preparation and encapsulation were performed at the pharmaceutical lab of the Department of Pharmaceutical Outcomes and Policy at the College of Pharmacy of the University of Florida under the auspices of the Florida Department of Health Internal Review Board. During the period of January 2014 to October 2016, C. Papaya capsules were administered to oncology patients to determine the effects on hematology and biochemical parameters. C. Papaya capsules were administered to 11 pediatric patients (males and females, 3-8 years of age, of various racial and ethnic backgrounds, and of various body composition) at the Loyola University Medical Center in Chicago Illinois (USA). C. Papaya capsules were administered to 95 adult patients (males and females, 18-72 years of age, of various racial and ethnic backgrounds, and of various body composition) with solid-tumor cancers (lung, breast) at the Loyola University Medical Center in Chicago Illinois (USA) or the Saint Bernard Hospital in Chicago Illinois (USA). C. Papaya capsules were administered to 10 adult Vietnamese patients (males and females, 18-72 years of age, and of various body composition) with solid-tumor cancer (tonsillar) at the International Medical Center in Ho Chi Minh City (Vietnam). Human consumption of C. Papaya capsules was supervised by licensed medical doctors and occurred under the auspices of the respective hospital’s Internal Review Boards. Hematologic and biochemical parameters were measured in the respective hospital’s clinical pathology laboratories using standard clinical methodologies.

Results and discussion

LC-MS analysis of C. Papaya leaf extract was used to detect the spectra (Figure 1), and 21 phyto-chemical constituents (Table 2) were identified. The presence of various pharmacologic phyto-compounds (alkaloids, phenolics, flavonoids, amino acids) was consistent with previously reported analyses of parts of C. Papaya specimens.

Figure 1a. LCMS spectrum (positive mode) analysis of extract of leaf of C. Papaya

Figure 1b. LMCS spectrum (negative mode) analysis of extract of leaf of C. Papaya

Table 2. Phytochemical analysis of extract of leaf of C. Papaya  

Number

Compound

Molecular Mass

1.

2.

3.

4.

5.

6.

7.

8.

9.

10.

11.

12.

13.

14.

15.

16.

17.

18.

19.

20.

21.

Tocopherol

Ascorbic acid

Carpaine

Deoxykaempfe

Kaempferol

Deoxyquercetin

Quercetin

Dicoumarol

Coumaroylquin

Coumarin

Folic acid

Cystine

Homocysteine

Cysteine

L Glutamic

p- C. Alcohol

dime- phenol

umbelliferone

phenylalanine

Caff-Alcohol

Methyl- ketone

420.67

173.05

455.76

267.41

278.67

280.34

300.45

340.42

344.57

147.67

440.65

120.32

133.59

176.33

143.33

152.44

153.67

160.11

164.59

165.02

169.79

The in vitro results shown in Figure 2 strongly suggest that C. Papaya leaf extract may inhibit tumor cell growth and proliferation and may promote apoptotic activity in tumor cells. Decreased 3[H]- TdR incorporation, resulting from exposure to increased concentrations of C. Papaya leaf extract, suggests a dose-dependent inhibition of tumor cell growth and proliferation in solid-tumor cells lines (Hela, MCF-7, HepG2, PC-14, Panc-1 and H2452) and large-cell lymphoma cell lines (Jurkat, ARH77, Raji and Karpas-299). Increased 7-Aminoactinomycin D (7-AAD) fluorescent marker and decreased Annexin V- cell detection after exposure to increased concentration of C. Papaya leaf extract suggest a dose-dependent promotion of apoptotic activity. The increased detection of luminescence after 0, 18 and 24-hour exposure to C. Papaya leaf extract or etoposide (positive control) demonstrates a dose-dependant promotion of apoptotic activity in these cancer cells.

Figure 2. Effects of C. Papaya leaf extract on tumor cell growth, proliferation and induction of apoptotic activity. (A) Fractional incorporation of 3H-thymidine (3[H]- TdR) in solid-tumor cell lines Hela, MCF-7, HepG2, PC-14, Panc-1 and H2452 at various concentrations of extract of C. Papaya leaf extract. Decreased 3[H]- TdR incorporation at higher extract concentrations demonstrates dose-dependent inhibition of tumor cell proliferation. (B) Fractional incorporation of 3H-thymidine (3[H]- TdR)TdR incorporation in large-cell lymphoma cell lines Jurkat, ARH77, Raji and Karpas-299 cells at various concentrations of extract of C. Papaya leaf extract. Decreased 3[H]- TdR incorporation at higher extract concentrations demonstrates dose-dependent inhibition of tumor cell proliferation. (C) 7-Aminoactinomycin D (7-AAD) fluorescent marker for detection of DNA from dead cells and labeled Annexin V- cells were used to measure apoptotic activity after exposure to C. Papaya leaf extract at various concentrations. (D) Luminescence after 0, 18 and 24-hour exposure to etoposide or C. Papaya leaf extract. Increased luminescence indicates increased apoptotic activity.

The in vitro results shown in Figure 3 strongly suggest that C. Papaya leaf extract may have important immune-modulatory effects. Decreased levels of inflammatory markers (IL-2, IL-4, IL-12 p40, IL-12 p70, INF-γ, and TNF-α) in human PBMCs following 24-hour exposure to increased concentrations of C. Papaya leaf extract demonstrate a dose-dependent attenuation of immune-oncologic activity.

Figure 3. Immuno-modulatory activity secondary to in vitro exposure to C. Papaya leaf extract. (A) Measured concentrations of a) IL-2 and b) IL-4 in human peripheral blood mononuclear cells (PBMCs) following 24-hour exposure to various concentrations of C. Papaya leaf extract. (B) a) Percent of annexin V-negative cells in human PBMCs following 24-hour exposure to C. Papaya leaf extract and b) fractional incorporation of 3H-thymidine (3[H]- TdR) in human PBMCs without and with 72-hour exposure to C. Papaya leaf extract. C) Measured concentrations of a) IL-12 p40, (b) IL-12 p70, (c) INF-γ, and (d) TNF- α in human PBMCs following 24-hour exposure to various concentrations of C. Papaya leaf extract.

The in vitro results shown in Figure 4 strongly suggest that C. Papaya leaf extract may have important cytotoxic effects. The decreased 3[H]- TdR detected and the increased percent lysis detected in human PBMCs after 24-hour exposure to C. Papaya leaf extract at increased effector-target ratios (E/T) demonstrate a dose-dependent cytotoxic effect in these cancer cells.

Figure 4. Cytotoxic effects of C. Papaya leaf extract. (A) fractional incorporation of 3H-thymidine (3[H]- TdR) in human peripheral blood mononuclear cells after 24-hour exposure to C. Papaya leaf extract at various effector-target ratios (E/T ratio). (B) Percent lysis of PBMCs at various E/T ratios. * denotes statistical significance (p < 0.05).

The in vitro results shown in Figure 5 strongly suggest that C. Papaya leaf extract may have important effects on genetic expression and regulation. Real-time PCR analysis revealed that increased mRNA expression in several cancer-related and/or immuno-regulatory genes (CCL2, CCL7, CCL8 and SERPINB2) resulted from exposure to C. Papaya leaf extract, and the results suggest that this effect on mRNA was dose-dependant. The clinical implications of the results are uncertain, and additional investigation is warranted.

Figure 5. Genetic expression and regulatory effects secondary to in vitro exposure to C. Papaya leaf extract. A) Real-time PCR analysis performed on peripheral blood mononuclear cells (PBMCs) without exposure (lane a) and with 24-hour exposure (lane b) to C. Papaya leaf extract. B) Genetic up-regulation of select genes in PBMCs after 24-hour exposure to C. Papaya leaf extract. C) Increased mRNA expression of select cancer biology related genes measured in PBMCs with and without 24-hour exposure to C. Papaya leaf extract. D) Dose-dependent increased mRNA expression of select cancer biology related genes measured in PBMCs without and with 24-hour exposure to C. Papaya leaf extract at low (0.06%) and high (0.125%) doses.

Human hematology laboratory measurements in cancer patients that received oral doses of encapsulated C. Papaya leaf extract are presented in Table 3. A decrease in MCV was observed in male patients treated with medium dose of C. Papaya leaf extract (p=0.043). The clinical importance of this finding is uncertain. Human clinical laboratory measurements (metabolic and organ system parameters) in cancer patients that received oral doses of C. Papaya leaf extract are presented in Table 4. Increased ALT levels were observed in male patients that received low and medium doses (p=0.032 and 0.029, respectively) of C. Papaya leaf extract. Increased Total Cholesterol was observed in female patient treated with medium doses (p=0.01).

Table 3. Hematologic parameters in male and female patients consuming various amounts of C. Papaya leaf extract. Values are expressed as mean+/-standard deviation. WBC: White Blood Cells; RBC: Red Blood Cells; HBG: Hemoglobin; MCV: Mean Corpuscular volume; MCHC: mean corpuscular hemoglobin concentration. * indicates a statistically significant (p<0.05) difference for control.​

Platelet (10K/mcL)

Gender

Hematologic
Parameter

Control

C. Papaya Leaf Extract
0.01g/kg BW

0 C. Papaya Leaf
Extract 0.16g/kg BW

C. Papaya Leaf
Extract 2.0 g/kg BW

Male

WBC (10K/mcL)

3.4+/-3.5

7.0+/-2.1

3.7+/-3.2

7.5+/-1.8

 

RBC (1000KxmcL)

6.5+/-0.3

6.8+/-0.3

7.2+/-0.2

7.0+/-0.2

 

Hb(g/dL)

13.0+/-0.5

14.4+/-0.7

14.0+/-0.3

14.6+/-0.6

 

MCV (fL)

63.8+/-0.8

63.4+/-0.4

*60.9+/-1.0

63.2+/-2.6

 

Lymphocyte (%)

83.4+/-6.5

*86.8+/-2.0

83.0+/-3.0

83.5+/-6.0

 

1012+/-221

1022+/-140

998+/-236

1022+/-130

 

MCHC (g/dL)

33.3+/-0.7

34.5+/-0.6

32.1+/-0.5

33.0+/-0.2

Female

WBC (10K/mcL)

6.8+/-2.0

5.9+/-0.8

4.2+/-3.4

6.5+/-1.7

 

RBC (1000KxmcL)

6.6+/-0.5

6.3+/-0.1

6.4+/-0.2

6.5+/-0.6

 

Hb(g/dL)

13.9+/-0.7

13.3+/-0.3

13.6+/-0.6

13.7+/-0.7

 

MCV (fL)

62.3+/-2.4

63.5+/-1.3

62.3+/-1.0

61.8+/-1.7

 

Lymphocyte (%)

81.6+/-10.1

84.6+/-1.7

83.2+/-4.9

82.0+/-5.3

 

Platelet (10K/mcL)

1030+/-434

1075+/-103

1153+/-300

1085+/-285

 

MCHC (g/dL)

34.0+/-0.8

34.5+/-0.2

22.0+/-0.3

20.2+/-0.6

Table 4. Clinical laboratory values in male and female patients consuming various amounts of C. Papaya leaf extract. Values are expressed as mean+/-standard deviation. AST: aspartate transaminase; ALT: Alanine Aminotransferase; CK: Creatinine Kinase; LDH: Lactate dehydrogenase. * indicates a statistically significant (p<0.05) difference for control.

Gender

Organ
Profile

Parameter

Control

C. Papaya Leaf
Extract
0.01g/kg BW

C. Papaya Leaf
Extract
0.16g/kg BW

C. Papaya Leaf
Extract
2.0 g/kg BW

Male

Metabolic

Glucose (mmol/L)

6.3+/-1.5

10.0+/-4.4

11.5+/-9.0

8.1+/-5.0

 

Liver

AST (U/L)

199+/-42

225+/-20

250+/-75

232+/-25

 

 

ALT (U/L)

58.2+/-12

82.0+/-16.0

*83.5+/-2.6

66.0+/-3.5

 

Renal

Creatinine (mcmol/L)

49.4+/-10.7

54.52+/-7.9

56.8+/-16.5

50.8+/-5.7

 

Cardiac

CK (U/L)

1021+/-190

1002+/-231

1189+/-675

740+/-196

 

 

LDH (U/L)

2505+/-340

2530+/-380

2070+/-560

2328+/-302

 

Lipid

Total Cholesterol (mmol/L)

1.3+/-0.2

1.3+/-0.2

1.5+/-0.1

1.4+/-0.3

Female

Metabolic

Glucose (mmol/L)

11.1+/-4.5

12.5+/-4.2

12.9+/-4.0

11.6+/-4.7

 

Liver

AST (U/L)

43.2+/-33

39.4+/-42

316+/-74

265+/-117

 

 

ALT (U/L)

52.5+/-22

61.4+/-4.8

89.0+/-7.0

73.5+/-40

 

Renal

Creatinine (mcmol/L)

64.2+/-4.2

70.4+/-10.3

65.4+/-3.5

50.0+/-5.9

 

Cardiac

CK (U/L)

1223+/-521

1209+/-423

1506+/-733

1223+/-570

 

 

LDH (U/L)

1768+/-820

1700+/-849

2169+/-805

2850+/-190

 

Lipid

Total Cholesterol (mmol/L)

1.7+/-0.3

1.8+/-0.3

*2.1+/-0.1

1.3+/-0.3

The clinical importance of these finding is uncertain. Oral consumption of C. Papaya leaf extract (capsules) was well tolerated: no adverse effects were observed and no patient complaints were reported.

Conclusions

Anecdotal reports along with the in vivo and in vitro studies presented in this report suggest that the leaf extract of C. Papaya has substantial potential as an adjuvant treatment for solid-tumors (e.g., lung, breast, tonsil). Some significant effects on certain hematologic and organ chemistry parameters were also observed. Additional investigation of C. Papaya leaf extract is warranted.

Acknowledgements

The authors are grateful to Ms. Gloria Gomez for assistance with data analysis. The authors are grateful for the assistance and expertise in laboratory analysis of the laboratory staff of the College of Pharmacy at the University of Florida (Gainseville, Florida, USA), the Pathology Department at Loyola University Medical Center (Chicago, Illinois, USA), and the Pathology Lab of Saint Bernard Hospital (Gainseville, Florida, USA). The authors are also thankful for the oversight of all aspects of this research but the Internal Review Boards of those institutions as well as the Florida Department of Health, the Illinois Department of Public Health and the Vietnam Board of Medicine and Pharmacy.

References

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Editorial Information

Editor-in-Chief

Terry Lichtor

Article Type

Research Article

Publication history

Received date: March 17, 2017
Accepted date: April 17, 2017
Published date: April 20, 2017

Copyright

© 2017 Nguyen H. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Citation

Nguyen H, Kitzmiller JP, Khungar P, Dabbade N, Nguyen KT, et al. (2017) Phytochemical, hematologic and anti-tumor activity evaluations of Carica papaya leaf extract. J Transl Sci 3: DOI: 10.15761/JTS.1000185

Corresponding author

Hoang Nguyen

Department of Biological Chemistry and Pharmacology, College of Medicine, The Ohio State University, Columbus, 5084 Graves Hall, 333 West Tenth Avenue, Columbus, OH 43210, USA

Figure 1a. LCMS spectrum (positive mode) analysis of extract of leaf of C. Papaya

Figure 1b. LMCS spectrum (negative mode) analysis of extract of leaf of C. Papaya

Figure 2. Effects of C. Papaya leaf extract on tumor cell growth, proliferation and induction of apoptotic activity. (A) Fractional incorporation of 3H-thymidine (3[H]- TdR) in solid-tumor cell lines Hela, MCF-7, HepG2, PC-14, Panc-1 and H2452 at various concentrations of extract of C. Papaya leaf extract. Decreased 3[H]- TdR incorporation at higher extract concentrations demonstrates dose-dependent inhibition of tumor cell proliferation. (B) Fractional incorporation of 3H-thymidine (3[H]- TdR)TdR incorporation in large-cell lymphoma cell lines Jurkat, ARH77, Raji and Karpas-299 cells at various concentrations of extract of C. Papaya leaf extract. Decreased 3[H]- TdR incorporation at higher extract concentrations demonstrates dose-dependent inhibition of tumor cell proliferation. (C) 7-Aminoactinomycin D (7-AAD) fluorescent marker for detection of DNA from dead cells and labeled Annexin V- cells were used to measure apoptotic activity after exposure to C. Papaya leaf extract at various concentrations. (D) Luminescence after 0, 18 and 24-hour exposure to etoposide or C. Papaya leaf extract. Increased luminescence indicates increased apoptotic activity.

Figure 3. Immuno-modulatory activity secondary to in vitro exposure to C. Papaya leaf extract. (A) Measured concentrations of a) IL-2 and b) IL-4 in human peripheral blood mononuclear cells (PBMCs) following 24-hour exposure to various concentrations of C. Papaya leaf extract. (B) a) Percent of annexin V-negative cells in human PBMCs following 24-hour exposure to C. Papaya leaf extract and b) fractional incorporation of 3H-thymidine (3[H]- TdR) in human PBMCs without and with 72-hour exposure to C. Papaya leaf extract. C) Measured concentrations of a) IL-12 p40, (b) IL-12 p70, (c) INF-γ, and (d) TNF- α in human PBMCs following 24-hour exposure to various concentrations of C. Papaya leaf extract.

Figure 4. Cytotoxic effects of C. Papaya leaf extract. (A) fractional incorporation of 3H-thymidine (3[H]- TdR) in human peripheral blood mononuclear cells after 24-hour exposure to C. Papaya leaf extract at various effector-target ratios (E/T ratio). (B) Percent lysis of PBMCs at various E/T ratios. * denotes statistical significance (p < 0.05).

Figure 5. Genetic expression and regulatory effects secondary to in vitro exposure to C. Papaya leaf extract. A) Real-time PCR analysis performed on peripheral blood mononuclear cells (PBMCs) without exposure (lane a) and with 24-hour exposure (lane b) to C. Papaya leaf extract. B) Genetic up-regulation of select genes in PBMCs after 24-hour exposure to C. Papaya leaf extract. C) Increased mRNA expression of select cancer biology related genes measured in PBMCs with and without 24-hour exposure to C. Papaya leaf extract. D) Dose-dependent increased mRNA expression of select cancer biology related genes measured in PBMCs without and with 24-hour exposure to C. Papaya leaf extract at low (0.06%) and high (0.125%) doses.

Table 1. Primary constituents of C. Papaya

Part

Constituents

Fruits

Fat, carbohydrates, phosphorous, iron, Vitamin C, thiamine, riboflavin, niacin, and carotene, amino acids, citric and malic acids (green fruits), volatile compounds: linalool, benzylisothiacyanate, protein, fat and fiber

Latex

Peptidase A and B and lysozymes, proteolytic enzymes, papain and chemopapain, glutamine cyclotransferase, chymopapains A, B and C

Leaves

Vitamin C and E, Alkaloids carpain, pseudocarpain and dehydracarpaine I and II, choline, carposide

Root

Enzyme myrosin and carposide

Table 2. Phytochemical analysis of extract of leaf of C. Papaya  

Number

Compound

Molecular Mass

1.

2.

3.

4.

5.

6.

7.

8.

9.

10.

11.

12.

13.

14.

15.

16.

17.

18.

19.

20.

21.

Tocopherol

Ascorbic acid

Carpaine

Deoxykaempfe

Kaempferol

Deoxyquercetin

Quercetin

Dicoumarol

Coumaroylquin

Coumarin

Folic acid

Cystine

Homocysteine

Cysteine

L Glutamic

p- C. Alcohol

dime- phenol

umbelliferone

phenylalanine

Caff-Alcohol

Methyl- ketone

420.67

173.05

455.76

267.41

278.67

280.34

300.45

340.42

344.57

147.67

440.65

120.32

133.59

176.33

143.33

152.44

153.67

160.11

164.59

165.02

169.79

Table 3. Hematologic parameters in male and female patients consuming various amounts of C. Papaya leaf extract. Values are expressed as mean+/-standard deviation. WBC: White Blood Cells; RBC: Red Blood Cells; HBG: Hemoglobin; MCV: Mean Corpuscular volume; MCHC: mean corpuscular hemoglobin concentration. * indicates a statistically significant (p<0.05) difference for control.​

Platelet (10K/mcL)

Gender

Hematologic
Parameter

Control

C. Papaya Leaf Extract
0.01g/kg BW

0 C. Papaya Leaf
Extract 0.16g/kg BW

C. Papaya Leaf
Extract 2.0 g/kg BW

Male

WBC (10K/mcL)

3.4+/-3.5

7.0+/-2.1

3.7+/-3.2

7.5+/-1.8

 

RBC (1000KxmcL)

6.5+/-0.3

6.8+/-0.3

7.2+/-0.2

7.0+/-0.2

 

Hb(g/dL)

13.0+/-0.5

14.4+/-0.7

14.0+/-0.3

14.6+/-0.6

 

MCV (fL)

63.8+/-0.8

63.4+/-0.4

*60.9+/-1.0

63.2+/-2.6

 

Lymphocyte (%)

83.4+/-6.5

*86.8+/-2.0

83.0+/-3.0

83.5+/-6.0

 

1012+/-221

1022+/-140

998+/-236

1022+/-130

 

MCHC (g/dL)

33.3+/-0.7

34.5+/-0.6

32.1+/-0.5

33.0+/-0.2

Female

WBC (10K/mcL)

6.8+/-2.0

5.9+/-0.8

4.2+/-3.4

6.5+/-1.7

 

RBC (1000KxmcL)

6.6+/-0.5

6.3+/-0.1

6.4+/-0.2

6.5+/-0.6

 

Hb(g/dL)

13.9+/-0.7

13.3+/-0.3

13.6+/-0.6

13.7+/-0.7

 

MCV (fL)

62.3+/-2.4

63.5+/-1.3

62.3+/-1.0

61.8+/-1.7

 

Lymphocyte (%)

81.6+/-10.1

84.6+/-1.7

83.2+/-4.9

82.0+/-5.3

 

Platelet (10K/mcL)

1030+/-434

1075+/-103

1153+/-300

1085+/-285

 

MCHC (g/dL)

34.0+/-0.8

34.5+/-0.2

22.0+/-0.3

20.2+/-0.6

Table 4. Clinical laboratory values in male and female patients consuming various amounts of C. Papaya leaf extract. Values are expressed as mean+/-standard deviation. AST: aspartate transaminase; ALT: Alanine Aminotransferase; CK: Creatinine Kinase; LDH: Lactate dehydrogenase. * indicates a statistically significant (p<0.05) difference for control.

Gender

Organ
Profile

Parameter

Control

C. Papaya Leaf
Extract
0.01g/kg BW

C. Papaya Leaf
Extract
0.16g/kg BW

C. Papaya Leaf
Extract
2.0 g/kg BW

Male

Metabolic

Glucose (mmol/L)

6.3+/-1.5

10.0+/-4.4

11.5+/-9.0

8.1+/-5.0

 

Liver

AST (U/L)

199+/-42

225+/-20

250+/-75

232+/-25

 

 

ALT (U/L)

58.2+/-12

82.0+/-16.0

*83.5+/-2.6

66.0+/-3.5

 

Renal

Creatinine (mcmol/L)

49.4+/-10.7

54.52+/-7.9

56.8+/-16.5

50.8+/-5.7

 

Cardiac

CK (U/L)

1021+/-190

1002+/-231

1189+/-675

740+/-196

 

 

LDH (U/L)

2505+/-340

2530+/-380

2070+/-560

2328+/-302

 

Lipid

Total Cholesterol (mmol/L)

1.3+/-0.2

1.3+/-0.2

1.5+/-0.1

1.4+/-0.3

Female

Metabolic

Glucose (mmol/L)

11.1+/-4.5

12.5+/-4.2

12.9+/-4.0

11.6+/-4.7

 

Liver

AST (U/L)

43.2+/-33

39.4+/-42

316+/-74

265+/-117

 

 

ALT (U/L)

52.5+/-22

61.4+/-4.8

89.0+/-7.0

73.5+/-40

 

Renal

Creatinine (mcmol/L)

64.2+/-4.2

70.4+/-10.3

65.4+/-3.5

50.0+/-5.9

 

Cardiac

CK (U/L)

1223+/-521

1209+/-423

1506+/-733

1223+/-570

 

 

LDH (U/L)

1768+/-820

1700+/-849

2169+/-805

2850+/-190

 

Lipid

Total Cholesterol (mmol/L)

1.7+/-0.3

1.8+/-0.3

*2.1+/-0.1

1.3+/-0.3