Sains Malaysiana 51(6)(2022): 1833-1845

http://doi.org/10.17576/jsm-2022-5106-19

The Effects of the Fractions of Piper sarmentosum Leaves on Inhibition of Adipogenesis of 3T3 L1 Preadipocytes

(Kesan daripada Fraksi Daun Piper sarmentosum pada Perencatan Adipogenesis pada Sel 3T3L1 Pra-adiposit)

FAIRUS AHMAD¹, IMA NIRWANA SOELAIMAN², KHAIRANA HUSAIN³, FARIHAH HJ SUHAIMI¹, CHUA KIEN HUI⁴ & NORZANA ABD GHAFAR^1,*

¹Department of Anatomy, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaakob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

²Department of Pharmacology, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaakob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

³Drug and Herbal Research Centre, Faculty of Pharmacy, Jalan Raja Muda Abdul Aziz, 53000 Kuala Lumpur, Federal Territory, Malaysia

⁴Department of Physiology, Universiti Kebangsaan Malaysia Medical Centre,

Jalan Yaakob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

Received: 29 June 2021/Accepted: 2 November 2021

ABSTRACT

Piper sarmentosum Roxb. was reported to have anti-obesity, hypoglycaemic and anti-oxidant properties. The aim of this study was to identify the fractions of P. sarmentosum leaf extract in inhibiting adipogenesis of 3T3L1 preadipocytes. The crude extract of the P. sarmentosum leaves was fractionated to produce hexane, dichloromethane, methanol, and aqueous fractions. Various dilutions of the fractions; hexane (0.1 - 1 µg/mL), dichloromethane (9.76 - 97.6 µg/mL), methanol (3.6 - 36 mg/mL), and aqueous (1 - 10 mg/mL), were treated onto the 3T3L1 preadipocytes from 3^rd to 15^th day of culture. The crude extract (1 - 10 mg/mL) and glycyrrhizic acid (GCA) (0.24 - 2.4 mg/mL) were used as positive controls. The viability of the adipocytes was measured by MTT assay at the 15^th day of culture. The content of each fraction was quantified with reference standards of naringin, naringenin, pellitorine, sarmentosine and β-sitosterol by using HPLC. The results showed that 49.1% of the crude extract contained aqueous fraction, 0.12% in hexane fraction, 9.7% in dichloromethane fraction and 36% in methanol fraction. The aqueous fraction and crude extract at the dose of 7 mg/mL and GCA at the dose of 1.92 mg/mL showed potent inhibitory effects on the adipogenesis. However, none of the reference standards were identified from the fractions using HPLC analysis. In conclusion, the aqueous fraction was the main fraction in the crude extract of the P. sarmentosum and contributed a significant role in inhibiting adipogenesis of the 3T3L1 preadipocytes.

Keywords: Adipogenesis; aqueous fraction; P. sarmentosum; 3T3L1 preadipocytes

Abstrak

Piper sarmentosum Roxb. adalah tumbuhan yang mempunyai ciri-ciri anti-obesiti, hipoglisemia dan antioksidan. Tujuan kajian ini adalah untuk mengenal pasti fraksi ekstrak daun P. sarmentosum dalam menghalang adipogenesis pada sel 3T3L1 pra-adiposit. Melalui proses fraksinasi, ekstrak daun P. sarmentosum ini dapat menghasilkan fraksi heksana, diklorometana, metanol dan akueus. Pencairan fraksi heksana (0.1 - 1 μg/mL), diklorometana (9.76 - 97.6 μg/mL), metanol (3.6 - 36 mg/mL) dan akueus (1 - 10 mg/mL) digunakan untuk merawat sel 3T3 L1 pra-adiposit dalam kultur. Pencairan ekstrak (1 - 10 mg/mL) dan asid gliserizik (GCA) (0.24 - 2.4 mg/mL) digunakan sebagai kawalan positif. Kebolehhidupan sel adiposit pada hari ke-15 kultur diukur dengan menggunakan kaedah asai MTT. Kandungan setiap fraksi dianalisis dengan menggunakan kaedah HPLC untuk mengesan rujukan sebatian piawai naringin, naringenin, pellitorine, sarmentosine dan β-sitosterol. Keputusan menunjukkan 49.1% ekstrak mengandungi fraksi akues, 0.12% fraksi heksana, 9.7% fraksi diklorometana dan 36% fraksi metanol. Fraksi akues dan ekstrak pada dos 7 mg/mL dan GCA pada dos 1.92 mg/mL menunjukkan kesan perencatan yang kuat terhadap adipogenesis. Walau bagaimanapun, tidak terdapat rujukan sebatian piawai pada fraksi ekstrak melalui analisis HPLC. Kesimpulannya, fraksi akues adalah fraksi utama ekstrak P. sarmentosum dan ia memainkan peranan yang penting dalam merencat adipogenesis sel 3T3L1 pra-adiposit.

Kata kunci: Adipogenesis; fraksi akues; P. sarmentosum; sel 3T3L1 pra-adiposit

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^*Corresponding author; email: norzana@ukm.edu.my